ABSTRACT
Objective: To study the effects of dihydromyricetin (DMY) on the proliferation, apoptosis and epithelial mesenchymal transition (EMT) of esophageal squamous cell carcinoma (ESCC) cell KYSE150 and KYSE410. Methods: KYSE150 and KYSE410 cells were treated with different concentrations of DMY (0, 25, 50, 100, 150, 200 µmol/L) for 24 hours. The median inhibition concentration (IC50) values of KYSE150 and KYSE410 were detected by cell counting kit-8 (CCK-8) method. Then 0.5 dimethyl sulfoxide (DMSO) was used as control group, dihydromyricetin (DMY), dihydromyricetin and transforming growth factor-ß1 (DMY+ TGF-ß1), transforming growth factor-ß1 (TGF-ß1) were used as experimental group. Cell proliferation and apoptosis rates were measured by clonal formation and flow cytometry. Transwell invasion and wound healing assay were used to detect cell invasion and migration. The protein expression levels of Caspase-3, Caspase-9, Bcl-2, Bax, Smad2/3, phosphorylation-Smad2/3 (p-Smad2/3) and Vimentin were detected by western blot. Results: The IC50 values of DMY on KYSE410 and KYSE150 cells were 100.51 and 101.27 µmol/L. The clone formation numbers of KYSE150 and KYSE410 in DMY group [(0.53±0.03) and (0.31±0.03)] were lower than those in DMSO group [(1.00±0.10) and (1.00±0.05), P<0.05]. The apoptosis rates of KYSE150 and KYSE410 cells in DMY group [(1.84±0.22)% and (2.80±0.07)%] were higher than those in DMSO group [(1.00±0.18)% and (1.00±0.07)%, P<0.05]. The invasion numbers of KYSE150 and KYSE410 cells in DMY group [(0.42±0.03) and (0.29±0.05)] were lower than those in DMSO group [(1.00±0.08) and (1.00±0.05), P<0.05]. The migration rates of KYSE150 and KYSE410 cells in DMY group [(0.65±0.14)% and (0.40±0.17)%] were lower than those in DMSO group [(1.00±0.10)% and (1.00±0.08)%, P<0.05]. The clone formation numbers of KYSE150 and KYSE410 in TGF-ß1 group [(1.01±0.08) and (0.99±0.25)] were higher than those in DMY+ TGF-ß1 group [(0.73±0.10) and (0.58±0.05), P<0.05]. The apoptosis rates of KYSE150 and KYSE410 cells in TGF-ß1 group [(0.81±0.14)% and (1.18±0.10)%] were lower than those in DMY+ TGF-ß1 group [(1.38±0.22)% and (1.85±0.04)%, P<0.05]. The invasion numbers of KYSE150 and KYSE410 cells in TGF-ß1 group [(1.19±0.11) and (1.39±0.11)] were higher than those in DMY+ TGF-ß1 group [(0.93±0.09) and (0.93±0.05), P<0.05]. The migration rates of KYSE150 and KYSE410 cells in TGF-ß1 group [(1.87±0.19)% and (1.32±0.04)%] were higher than those in DMY+ TGF-ß1 group [(0.86±0.16)% and (0.77±0.12)%, P<0.05]. The protein expression levels of Bax, Caspase-3 and Caspase-9 in KYSE150 and KYSE410 cells in DMY group were higher than those in DMSO group, while the protein expression level of Bcl-2 was lower than that in DMSO group (P<0.05). The protein expression levels of p-Smad2/3, Smad2/3 and Vimentin in KYSE150 and KYSE410 cells in DMY group were lower than those in DMSO group (P<0.05). The protein expression levels of Bax, Caspase-3 and Caspase-9 in KYSE150 and KYSE410 cells in TGF-ß1 group were lower than those in DMY+ TGF-ß1 group, and the protein expression level of Bcl-2 was higher than that in DMY+ TGF-ß1 group (P<0.05). The protein expression levels of Bax, Caspase-3 and Caspase-9 in KYSE150 and KYSE410 cells in DMY+ TGF-ß1 group were lower than those in DMY group, and the protein expression level of Bcl-2 was higher than that in DMY group (P<0.05). The protein expression levels of p-Smad2/3, Smad2/3 and Vimentin in KYSE150 and KYSE410 cells in TGF-ß1 group were higher than those in DMY+ TGF-ß1 group (P<0.05). Conclusion: DMY can inhibit the proliferation and EMT of ESCC mediated by TGF-ß1 and promote cell apoptosis.
Subject(s)
Esophageal Neoplasms , Esophageal Squamous Cell Carcinoma , Apoptosis , Caspase 3/metabolism , Caspase 9/metabolism , Cell Line, Tumor , Cell Movement , Cell Proliferation , Dimethyl Sulfoxide/pharmacology , Epithelial-Mesenchymal Transition , Esophageal Neoplasms/metabolism , Flavonols , Humans , Signal Transduction , Transforming Growth Factor beta1/metabolism , Transforming Growth Factor beta1/pharmacology , Vimentin/metabolism , bcl-2-Associated X Protein/metabolism , bcl-2-Associated X Protein/pharmacologyABSTRACT
This study investigated the association of tumor necrosis factor-α (TNF-α)-308, -238, and -863 polymorphisms with osteoarticular tuberculosis (OA-TB) prognosis in a Hebei population. Genomic DNA was extracted from venous blood samples of 120 OA-TB patients and 100 healthy volunteers. TNF-α-308, -238, and -863 were analyzed by PCR-restriction fragment length polymorphism; genotype and allele frequencies were calculated. Serum TNF-α level was significantly higher in OA-TB patients (283.16 ± 51.68 ng/L) than in control (122.54 ± 54.65 ng/L; P < 0.05). Higher frequency of TNF-α-308 GG genotype in healthy volunteers (91.0%) than in OA-TB patients (79.2%) indicated that it was a protective factor against OA-TB (OR = 0.405, 95%CI = 0.147-0.657, P = 0.007). Higher frequencies of TNF-α-308 GA genotype and TNF-α-308 allele (A) in OA-TB patients (20.8 and 10.4%, respectively) than in healthy volunteers (8.0 and 5.0%, respectively) indicated an association with increased risk of OA-TB (OR = 3.112, 95%CI = 1.520-6.343, P = 0.003; OR = 3.109, 95%CI = 1.676-6.538, P = 0.006; respectively). Haplotype association analysis of TNF-α polymorphisms (-308/-238/-863) showed a higher frequency of TNF-α AGA in OA-TB patients (12.1%) than in healthy volunteers (3.5%), indicating that it was a risk factor for OA-TB (OR = 4.201, 95%CI = 1.80-9.91, P = 0.010). TNF-α-308 G/A and TNF-α AGA (-308/-238/-863) were associated with a predisposition to OA-TB, which could aid clinical detection, prevention, and prognosis of OA-TB.
Subject(s)
Genetic Association Studies , Genetic Predisposition to Disease , Tuberculosis, Osteoarticular/genetics , Tumor Necrosis Factor-alpha/genetics , Adult , Alleles , Female , Genotype , Haplotypes , Humans , Male , Middle Aged , Polymorphism, Single Nucleotide , Prognosis , Risk Factors , Tuberculosis, Osteoarticular/pathologyABSTRACT
This study aimed to analyze the expression, clinical significance of cyclin G2 (CCNG2) in colorectal carcinoma, and the biological effect in its cell line by CCNG2 overexpression. Immunohistochemistry and Western blot were used to analyze CCNG2 protein expression in colorectal cancer and to study the influence of the upregulated expression of CCNG2 that might be found on SW480 cell biological effect. We found that the level of CCNG2 protein expression was significantly lower in colorectal cancer tissue than normal tissues (P < 0.05). The level of CCNG2 was correlated with T stages, lymph node metastasis, clinic stage, and histological grade (P < 0.05). Loss of CCNG2 expression correlated significantly with poor overall survival time by Kaplan-Meier analysis (P < 0.05). The result of biological function has shown that SW480 cell-transfected CCNG2 had a lower survival fraction, higher percentage of the G0/G1 phases, and lower CDK2 protein expression compared with SW480 cell-untransfected CCNG2 (P < 0.05).
Subject(s)
Colorectal Neoplasms/pathology , Cyclin G2/physiology , Adult , Aged , Cell Cycle , Colorectal Neoplasms/metabolism , Colorectal Neoplasms/mortality , Cyclin G2/analysis , Cyclin G2/genetics , Cyclin-Dependent Kinase 2/physiology , Down-Regulation , Female , Humans , Male , Middle Aged , Neoplasm Staging , PrognosisABSTRACT
This study aims to analyze the expression and clinical significance of Filamin A (FLNA) in prostate carcinoma and the biological effect in its cell line by FLNA overexpression. Immunohistochemistry and Western blot were used to analyze FLNA protein expression in 68 cases of prostate cancer and 37 cases of normal tissues to study the influence of the upregulated expression of FLNA that might be found on PC-3 cell biological effect. In the immunohistochemical analysis, the level of FLNA protein expression was found to be significantly lower in prostate cancer tissue than in normal tissues (P < 0.05). In the Western blot analysis, the relative amount of FLNA protein in prostate cancer tissue was found to be significantly lower than in normal tissues (P < 0.05). The level of FLNA protein expression was not correlated with age and PSA concentration (P > 0.05), but it was correlated with T stages, lymph node metastasis, clinic stage, and Gleason score (P < 0.05). The result of biological function showed that PC-3 cell transfected FLNA had a lower survival fraction, a significant decrease in migration and invasion, and a lower matrix metallopeptidase 9 (MMP-9) protein expression compared with PC-3 cell untransfected FLNA (P < 0.05). FLNA expression decreased in prostate cancer and correlated significantly with T stages, lymph node metastasis, clinic stage, and Gleason score, suggesting that FLNA may play important roles as a negative regulator to prostate cancer PC-3 cell by promoting the degradation of MMP-9.
Subject(s)
Cell Movement , Filamins/physiology , Matrix Metalloproteinase 9/metabolism , Prostatic Neoplasms/pathology , Aged , Filamins/analysis , Filamins/genetics , Humans , Immunohistochemistry , Lymphatic Metastasis , Male , Middle Aged , Neoplasm Invasiveness , Neoplasm Staging , Prostatic Neoplasms/enzymologyABSTRACT
This study aimed to analyze the expression, clinical significance of B cell translocation gene 1 (BTG1) in nonsmall cell lung cancer (NSCLC) and the biological effect in its cell line by BTG1 overexpression. Immunohistochemistry and western blot were used to analyze BTG1 protein expression in 82 cases of NSCLC and 38 cases of normal tissues to study the relationship between BTG1 expression and clinical factors. Recombinant lentiviral vector was constructed to overexpress EMP-1 and then infect NSCLC H1299 cell line. Quantitative real-time RT-PCR and western blot were used to detect the mRNA level and protein of BTG1. 3-[4,5-dimethylthiazol -2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, cell apoptosis, cell cycles, and migration and invasion assays were also conducted as to the influence of the upregulated expression of BTG1 that might be found on H1299 cells biological effect. The level of BTG1 protein expression was found to be significantly lower in NSCLC tissue than normal tissues (P < 0.05). Decreased expression of BTG1 was significantly correlated with lymph node metastasis, clinic stage, and histological grade of patients with NSCLC (P < 0.05). Meanwhile, loss of BTG1 expression correlated significantly with poor overall survival time by Kaplan-Meier analysis (P < 0.05). The result of biological function show that H1299 cell transfected BTG1 had a lower survival fraction; higher percentage of the G0/G1 phases; higher cell apoptosis; significant decrease in migration and invasion; and lower CyclinD1, Bcl-2, and MMP-9 protein expression compared with H1299 cell untransfected BTG1 (P < 0.05). BTG1 expression decreased in NSCLC and correlated significantly with lymph node metastasis; clinical stage; histological grade; poor overall survival; cell proliferation; cell cycles; cell apoptosis; and migration and invasion in NSCLC cell by regulating CyclinD1, Bcl-2, and MMP-9 protein expression, suggesting that BTG1 may play important roles as a negative regulator to NSCLC cell.
Subject(s)
Carcinoma, Non-Small-Cell Lung/pathology , Lung Neoplasms/pathology , Neoplasm Proteins/physiology , Adult , Aged , Carcinoma, Non-Small-Cell Lung/mortality , Carcinoma, Non-Small-Cell Lung/secondary , Cell Line, Tumor , Cell Proliferation , Down-Regulation , Female , Humans , Lung/chemistry , Lung Neoplasms/mortality , Male , Middle Aged , Neoplasm Invasiveness , Neoplasm Proteins/analysis , Neoplasm Proteins/genetics , PrognosisABSTRACT
This study aimed to analyze the expression, clinical significance of cyclin G2 (CCNG2) in gastric carcinoma, and the biological effect in its cell line by CCNG2 overexpression. Immunohistochemistry and western blot were used to analyze CCNG2 protein expression in 87 cases of gastric cancer and normal tissues to study on the relationship between CCNG2 expression and clinical factors. CCNG2 lentiviral vector was transfected into gastric SGC-7901 cell line. RT-PCR and western blot were used to detect the mRNA level and protein of CCNG2. MTT assay and cell cycle were also conducted as to the influence of the upregulated expression of CCNG2 that might be found on SGC-7901 cell biological effect. Immunohistochemically, the level of CCNG2 protein expression was found to be significantly lower in gastric cancer tissue than in normal tissues (P < 0.05). Western blot shows that the relative amount of CCNG2 protein in gastric cancer tissue was found to be significantly lower than in normal tissues (P < 0.05). The level of CCNG2 protein expression was correlated with T stages, lymph node metastasis, clinical stage, and histological grade (P < 0.05). Loss of CCNG2 expression correlated significantly with poor overall survival time by Kaplan-Meier analysis (P < 0.05). The result of biological function showed that SGC-7901 cell-transfected CCNG2 had a lower survival fraction, higher percentage of the G0/G1 phases, and lower CDK2 protein expression compared with SGC-7901 cell untransfected CCNG2 (P < 0.05). CCNG2 expression decreased in gastric cancer and correlated significantly T stages, lymph node metastasis, clinical stage, histological grade, and poor overall survival, suggesting that CCNG2 may play important roles as a negative regulator to gastric cancer cell by promoting degradation of CDK2.
Subject(s)
Adenocarcinoma/metabolism , Adenocarcinoma/pathology , Cell Transformation, Neoplastic/metabolism , Cyclin G2/metabolism , Stomach Neoplasms/metabolism , Stomach Neoplasms/pathology , Adenocarcinoma/mortality , Adult , Aged , Blotting, Western , Cell Line, Tumor , Female , Flow Cytometry , Humans , Immunohistochemistry , Kaplan-Meier Estimate , Male , Middle Aged , Neoplasm Grading , Neoplasm Staging , Prognosis , Reverse Transcriptase Polymerase Chain Reaction , Stomach Neoplasms/mortality , Transfection , Young AdultABSTRACT
This study aimed to analyze the expression and clinical significance of filamin A (FLNA) in gastric carcinoma and the biological effect in its cell line by FLNA overexpression. Immunohistochemistry and western blot were used to analyze FLNA protein expression in 47 cases of gastric cancer and 47 cases of normal tissues to study the relationship between FLNA expression and clinical factors. FLNA lentiviral vector and empty vector were respectively transfected into gastric cancer SGC-7901 cell line. Reverse transcription-polymerase chain reaction (RT-PCR) and western blot were used to detect the mRNA level and protein of FLNA. 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and migration and invasion assays were also conducted to determine the influence of the upregulated expression of FLNA that might be found on SGC-7901 cell biological effect. Immunohistochemistry: The level of FLNA protein expression was found to be significantly lower in gastric cancer tissue than normal tissues (P < 0.05). Western blot: The relative amount of FLNA protein in gastric cancer tissue was found to be significantly lower than in normal tissues (P < 0.05). The level of FLNA protein expression was not correlated with gender, age, and tumor invasion (P > 0.05), but it was correlated with lymph node metastasis, clinic stage, and histological grade (P < 0.05). Loss of FLNA expression correlated significantly with poor overall survival time by Kaplan-Meier analysis (P < 0.05). The result of biological function showed that SGC-7901 cell transfected FLNA had a lower survival fraction, significant decrease in migration and invasion, and lower matrix metallopeptidase 9 (MMP-9) protein expression compared with SGC-7901 cell untransfected FLNA (P < 0.05). FLNA expression decreased in gastric cancer and correlated significantly with lymph node metastasis, clinic stage, histological grade, and poor overall survival, suggesting that FLNA may play important roles as a negative regulator to gastric cancer SGC-7901 cell by promoting degradation of MMP-9.
Subject(s)
Adenocarcinoma/metabolism , Biomarkers, Tumor/analysis , Cell Movement , Filamins/biosynthesis , Stomach Neoplasms/metabolism , Adenocarcinoma/mortality , Adenocarcinoma/pathology , Adult , Aged , Blotting, Western , Cell Line, Tumor , Cell Movement/genetics , Female , Filamins/genetics , Humans , Immunohistochemistry , Kaplan-Meier Estimate , Lymphatic Metastasis , Male , Middle Aged , Neoplasm Grading , Neoplasm Invasiveness/genetics , Neoplasm Staging , Prognosis , Reverse Transcriptase Polymerase Chain Reaction , Stomach Neoplasms/mortality , Stomach Neoplasms/pathology , Transfection , Young AdultABSTRACT
This study aimed to analyze the expression, clinical significance of filamin A (FLNA) in nasopharyngeal carcinoma, and the biological effect in its cell line by FLNA overexpression. Immunohistochemistry and western blot were used to analyze FLNA protein expression in 63 cases of nasopharyngeal cancer and 21 cases of normal tissues to study the relationship between FLNA expression and clinical factors. FLNA lentiviral vector and empty vector were respectively transfected into nasopharyngeal cancer CNE2 cell line. Reverse transcription-polymerase chain reaction (RT-PCR) and western blot were used to detect the mRNA level and protein of FLNA. 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide, migration, and invasion assays were also conducted as to the influence of the upregulated expression of FLNA that might be found on CNE2 cell biological effect. Immunohistochemistry: the level of FLNA protein expression was found to be significantly lower in nasopharyngeal cancer tissue than normal tissues (P < 0.05). Western blot: the relative amount of FLNA protein in nasopharyngeal cancer tissue was found to be significantly lower than in normal tissues (P < 0.05). The level of FLNA protein expression was correlated with T stages, lymph node metastasis, clinic stage, and histological grade (P < 0.05). Loss of FLNA expression correlated significantly with poor overall survival time by Kaplan-Meier analysis (P < 0.05). The result of biological function has shown that CNE2 cell-transfected FLNA had a lower survival fraction, significant decrease in migration and invasion, and lower matrix metallopeptidase 9 (MMP-9) protein expression compared with CNE2 cell-untransfected FLNA (P < 0.05). FLNA expression decreased in nasopharyngeal cancer and correlated significantly lymph node metastasis, clinic stage, histological grade, and poor overall survival, suggesting that FLNA may play important roles as a negative regulator to nasopharyngeal cancer CNE2 cell by promoting degradation of MMP-9.
Subject(s)
Filamins/physiology , Nasopharyngeal Neoplasms/mortality , Carcinoma , Cell Movement , Filamins/analysis , Humans , MAP Kinase Signaling System , Matrix Metalloproteinase 9/metabolism , Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms/chemistry , Nasopharyngeal Neoplasms/genetics , Nasopharyngeal Neoplasms/pathology , Neoplasm Invasiveness , PrognosisABSTRACT
This study aimed to analyze the expression, clinical significance of epithelial membrane protein 1 (EMP1) in breast carcinoma and the biological effect in its cell line by EMP1 overexpression. Immunohistochemistry and western blot were used to analyze EMP1 protein expression in 67 cases of breast cancer and 35 cases of normal tissues to study the relationship between EMP1 expression and clinical factors. Quantitative real-time RT-PCR and western blot were used to detect the mRNA level and protein of EMP1. MTT assay, migration and invasion assays were also conducted as to the influence of the upregulated expression of EMP1 that might be found on MCF-7 cell biological effect. The relative amount of EMP1 protein in breast cancer tissue was found to be significantly lower than in normal tissues (P < 0.05). The level of EMP1 protein expression was correlated with T stages, lymph node metastasis, clinic stage, and histological grade (P < 0.05). Meanwhile, loss of EMP1 expression correlated significantly with poor overall survival time by Kaplan-Meier analysis (P < 0.05). The result shown that MCF-7 cell transfected EMP1 had a lower survival fraction, higher cell apoptosis, significant decrease in migration and invasion, higher caspase-9, and lower VEGFC protein expression compared with MCF-7 cell untransfected EMP1 (P < 0.05). EMP1 expression decreased in breast cancer and correlated significantly with lymph node metastasis, clinic stage, histological grade, and poor overall survival, T stages, suggesting that EMP1 may play important roles as a negative regulator to breast cancer MCF-7 cell by regulating the expression of caspase 9 and VEGFC protein.
Subject(s)
Breast Neoplasms/pathology , Neoplasm Proteins/physiology , Receptors, Cell Surface/physiology , Breast/chemistry , Breast Neoplasms/mortality , Caspase 9/analysis , Cell Proliferation , Female , Humans , Lymphatic Metastasis , MCF-7 Cells , Neoplasm Proteins/analysis , Neoplasm Proteins/genetics , Neoplasm Staging , Prognosis , Receptors, Cell Surface/analysis , Receptors, Cell Surface/genetics , Vascular Endothelial Growth Factor C/analysisABSTRACT
To determine the expression and function of B cell translocation gene 1 (BTG1) in esophageal carcinoma, esophageal samples were taken from cancer lesions (n = 74) and adjacent normal tissue (n = 34) in esophageal cancer patients immediately after endoscopic biopsy. BTG1 expression was determined by immunohistochemistry and Western blotting. The effect of BTG1 overexpression was examined in vitro utilizing a human esophageal cancer cell line ECA-109 stably transfected with a recombinant lentivirus (LeBTG1 cells) and compared to empty vector-transfected controls (LeEmpty). BTG1 overexpression was verified by real-time reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot. The expression of proteins involved in cell cycle regulation (cyclin D1) and apoptosis (Bcl-2) and cell migration (MMP-9) in LeBTG1 cells was analyzed by Western blot. The effect of BTG1 overexpression on cell viability and proliferation was assessed by an MTT assay in LeBTG1 and LeEmpty cells. Flow cytometric analyses were used to evaluate the effect of BTG1 expression on cell cycle distribution and apoptosis. The migration and invasion potential of LeBTG1 cells was examined by plating cells in Matrigel-coated chambers. The level of BTG1 protein expression was found to be significantly lower in esophageal cancer tissue than normal tissues (P < 0.05). Decreased expression of BTG1 was significantly correlated with lymph node metastasis, clinical stage, and histological grade of patients with esophageal cancer (P < 0.05). Meanwhile, loss of BTG1 expression correlated significantly with poor overall survival time by Kaplan-Meier analysis (P < 0.05). The result of biological function shown that Eca-109 cell-transfected BTG1 had a lower survival fraction, higher percentage of the G0/G1 phases, higher cell apoptosis, significant decrease in migration and invasion, and lower cylin D1, Bcl-2, and MMP-9 protein expression compared with Eca-109 cell-untransfected BTG1 (P < 0.05). Reduced BTG1 expression is associated with increased disease severity, suggesting it is a negative regulator of esophageal cancer and can serve as a prognostic indicator.
Subject(s)
Biomarkers, Tumor/analysis , Carcinoma, Squamous Cell/metabolism , Carcinoma, Squamous Cell/pathology , Esophageal Neoplasms/metabolism , Esophageal Neoplasms/pathology , Neoplasm Proteins/biosynthesis , Aged , Apoptosis , Carcinoma, Squamous Cell/mortality , Cell Proliferation , Down-Regulation , Esophageal Neoplasms/mortality , Esophageal Squamous Cell Carcinoma , Female , Flow Cytometry , Humans , Immunohistochemistry , Kaplan-Meier Estimate , Male , Middle Aged , Neoplasm Invasiveness/pathology , Neoplasm Proteins/analysis , Prognosis , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
This study aimed to analyze the expression, clinical significance of f epithelial membrane protejn-1 (EMP-1) in prostate carcinoma, and the biological effect in its cell line by EMP1 overexpression. Immunohistochemistry and Western blot were used to analyze EMP1 protein expression in 76 cases of prostate cancer and 34 cases of normal tissues to study the relationship between EMP1 expression and clinical factors. EMP1 lentiviral vector and empty vector were respectively transfected into prostate cancer PC-3 cell line. Quantitative real-time reverse transcription polymerase chain reaction and Western blot were used to detect the mRNA level and protein of EMP1. 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, migration, and invasion assays were also conducted as to the influence of the upregulated expression of EMP1 that might be found on PC-3 cell biological effect. Immunohistochemistry: The level of EMP1 protein expression was found to be significantly lower in prostate cancer tissue than normal tissues (P < 0.05). Western blot: The relative amount of EMP1 protein in prostate cancer tissue was found to be significantly lower than in normal tissues (P < 0.05). The level of EMP1 protein expression was not correlated with age and prostate-specific antigen (PSA) concentration (P > 0.05), but it was correlated with T stages, lymph node metastasis, clinic stage, and Gleason score (P < 0.05). The result of biological function shown that PC-3 cell transfected EMP1 had a lower survival fraction, higher cell apoptosis, significant decrease in migration and invasion, higher caspase-9, and lower VEGFC protein expression compared with PC-3 cell untransfected EMP1 (P < 0.05). EMP1 expression decreased in prostate cancer and correlated significantly T stages, lymph node metastasis, clinic stage, and Gleason score, suggesting that EMP1 may play important roles as a negative regulator to prostate cancer PC-3 cell by regulating the expression of regulation of caspase-9 and VEGFC protein.
Subject(s)
Caspase 9/analysis , Neoplasm Proteins/physiology , Prostatic Neoplasms/pathology , Receptors, Cell Surface/physiology , Vascular Endothelial Growth Factor C/analysis , Adult , Aged , Cell Line, Tumor , Cell Proliferation , Humans , Male , Middle Aged , Neoplasm Invasiveness , Neoplasm Proteins/analysis , Neoplasm Proteins/genetics , Neoplasm Staging , Prostatic Neoplasms/chemistry , Receptors, Cell Surface/analysis , Receptors, Cell Surface/geneticsABSTRACT
This study aimed to analyze the expression, clinical significance of epithelial membrane protein-1 (EMP1) in nasopharyngeal carcinoma, and the biological effect in its cell line by EMP1 overexpression. Immunohistochemistry and Western blot were used to analyze the EMP1 protein expression in 75 cases of nasopharyngeal cancer and 31 cases of normal tissues to study the relationship between EMP1 expression and clinical factors. Recombinant lentiviral vector was constructed to overexpress EMP1 and then infect nasopharyngeal cancer CNE2 cell line. Quantitative real-time RT-PCR and Western blot were used to detect the mRNA level and protein of EMP1. MTT assay, cell apoptosis, migration, and invasion assays were also conducted to determine the influence of the upregulated expression of EMP1 that might be found on CNE2 cells' biological effect. Immunohistochemistry and Western blot: The level of EMP1 protein expression was found to be significantly lower in nasopharyngeal cancer tissue than in the normal tissues (P < 0.05). Decreased expression of EMP1 was significantly correlated with T stages, lymph node metastasis, clinic stage, and histological grade of patients with nasopharyngeal cancer (P < 0.05). Meanwhile, the loss of EMP1 expression correlated significantly with poor overall survival time by Kaplan-Meier analysis (P < 0.05). The result of biological function has shown that CNE2 cell-transfected EMP1 had a lower survival fraction, higher cell apoptosis, significant decrease in migration and invasion, higher caspase-9, and lower vascular endothelial growth factor C protein expression compared with CNE2 cell-untransfected EMP1 (P < 0.05). EMP1 expression decreased in nasopharyngeal cancer and correlated significantly T stages, lymph node metastasis, clinic stage, histological grade, and poor overall survival, suggesting that EMP1 may play important roles as a negative regulator to nasopharyngeal cancer cell.
Subject(s)
Apoptosis , Cell Proliferation , Nasopharyngeal Neoplasms/pathology , Neoplasm Proteins/physiology , Neovascularization, Pathologic/prevention & control , Receptors, Cell Surface/physiology , Adult , Aged , Female , Humans , Lymphatic Metastasis , Male , Middle Aged , Nasopharyngeal Neoplasms/blood supply , Nasopharyngeal Neoplasms/mortality , Nasopharyngeal Neoplasms/therapy , Neoplasm Proteins/analysis , Neoplasm Staging , Prognosis , Receptors, Cell Surface/analysis , Vascular Endothelial Growth Factor C/analysisABSTRACT
To determine the expression and function of B cell translocation gene 1 (BTG1) in nasopharyngeal carcinoma. Nasopharyngeal samples were taken from cancer lesions (n = 75) and adjacent normal tissue (n = 33) in nasopharyngeal cancer patients immediately after endoscopic biopsy. BTG1 expression was determined by immunohistochemistry and Western blotting. The effect of BTG1 overexpression was examined in vitro utilizing a human nasopharyngeal cancer cell line CNE2 stably transfected with a recombinant lentivirus (LeBTG1 cells) and compared to empty vector-transfected controls (LeEmpty). BTG1 overexpression was verified by real-time reverse transcriptase polymerase chain reaction and Western blot. The expression of proteins involved in cell cycle regulation (cyclin D1), apoptosis (Bcl-2) and cell migration (MMP-9) in LeBTG1 cells were analyzed by Western blot. The effect of BTG1 overexpression on cell viability and proliferation was assessed by an MTT assay in LeBTG1 and LeEmpty cells. Flow cytometric analyses were used to evaluate the effect of BTG1 expression on cell cycle distribution and apoptosis. The migration and invasion potential of LeBTG1 cells was examined by plating cells in Matrigel-coated chambers. BTG1 protein expression was significantly lower in nasopharyngeal cancer tissue biopsies than normal tissue as measured by immunohistochemistry (36.0 vs. 81.8 % of tissues; P < 0.05) and Western blotting (0.221 ± 0.019 vs. 0.652 ± 0.055; P < 0.05). Decreased expression of BTG1 was significantly correlated with nasopharyngeal cancer tumor stage, lymph node metastasis, clinical stage and pathologic differentiation (P < 0.05), as well as with reduced overall five-year survival rates compared to patients with higher expression levels (31.2 vs. 70.2 %; P < 0.05). In vitro analyses revealed that LeBTG1 cells had a reduced survival fraction compared to control LeEmpty cells, with higher rates of apoptosis (9.3 ± 0.7 vs. 2.3 ± 0.3 %; P < 0.05). The proportion of LeBTG1 cells in G0/G1 stage and S phase was also significantly different from LeEmpty cells (82.6 ± 3.8 and 10.1 ± 1.0 %, vs. 62.2 ± 2.4 and 28.9 ± 2.0 %, respectively; Ps < 0.05), and the migration and invasion of LeBTG1 cells was significantly impaired with respect to LeEmpty cells (96.0 ± 13.0 and 91.0 ± 11.0 vs. 158.0 ± 17.0 and 142.0 ± 15.0, respectively; Ps < 0.05). These effects were accompanied by decreased protein expression of cyclin D1, Bcl-2 and MMP-9 in LeBTG1 cells (0.231 ± 0.021, 0.413 ± 0.046, 0.131 ± 0.011, respectively) compared to control LeEmpty cells (0.636 ± 0.067, 0.821 ± 0.083, 0.451 ± 0.041, respectively; Ps < 0.05). Reduced BTG1 expression is associated with increased disease severity, suggesting it is a negative regulator of nasopharyngeal cancer and can serve as a prognostic indicator.
Subject(s)
Down-Regulation , Gene Expression Regulation, Neoplastic , Nasopharyngeal Neoplasms/genetics , Neoplasm Proteins/genetics , Adult , Aged , Apoptosis/genetics , Biomarkers, Tumor , Carcinoma , Cell Cycle/genetics , Cell Movement/genetics , Collagen , Cyclin D1/genetics , Drug Combinations , Humans , Laminin , Lymphatic Metastasis , Matrix Metalloproteinase 9/genetics , Middle Aged , Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms/pathology , Proteoglycans , Proto-Oncogene Proteins c-bcl-2/geneticsABSTRACT
BACKGROUND: Epidemiological and experimental carcinogenesis studies provide evidence that components of garlic (Allium sativum) have anticancer activity. Hepatocellular carcinoma is highly malignant and metastatic. Currently, there is no effective chemotherapy for patients with advanced Hepatocellular carcinoma leading to an urgent need to seek for novel therapeutic options. AIM: To investigate the effect of cell growth, cell apoptosis of Garlic-Derived Compound S-Allylmercaptocysteine (SAMC) on Human Gastric Cancer Cells Line SGC 7901 cells. MATERIALS AND METHODS: The SGC 7901 cells were cultured with different concentration's SAMC. Cell viability was detected by AO/EB staining. JNK and P38 pathway were assayed by PCR (polymerase chain reaction). RESULTS: The best concentration of SAMC (300 µM) for induction SGC 7901 apoptosis was confirmed through cell viability. The PCR assay demonstrated that JNK and P38 pathway play important role in apoptosis of SGC 7901 cells. CONCLUSIONS: This study indicated that SAMC can inhibit cell proliferation and induct apoptosis of SGC 7901 cells via JNK and P38 pathway.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis/drug effects , Cysteine/analogs & derivatives , Stomach Neoplasms/drug therapy , Cell Line, Tumor , Cysteine/pharmacology , Garlic/chemistry , Humans , MAP Kinase Signaling System/drug effects , Stomach Neoplasms/enzymology , Stomach Neoplasms/pathologyABSTRACT
Objective: To evaluate the early efficacy and safety of transnasal endoscopic decompression in the annulus of zinn (AZ) region for refractory dysthyroid optic neuropathy (DON) and to preliminarily analyze the correlated factors of postoperative visual function outcome. Methods: From July 2021 to January 2023, 35 patients (56 eyes) with DON who received AZ area decompression in Peking University Third Hospital were included retrospectively, including 9 males (13 eyes) and 26 females (43 eyes), aging (52.2±12.0) years. Among them, 35 eyes underwent two-wall (medial and inferior) orbital decompression using an endonasal endoscopic approach, while 21 eyes received three-wall (medial, lateral, and inferior) orbital decompression through a combined approach. Key parameters such as best corrected visual acuity (BCVA), visual field (MD value), eyeball prominence, intraocular pressure, and complications were recorded. Postoperative data were collected one month after surgery. The statistical analysis was performed using paired t-test and Spearman correlation analysis. Results: Significant outcomes were observed post surgery in BCVA, visual field, intraocular pressure and proptosis (t value was 8.37, 6.17, 4.50, and 9.20, respectively, all P<0.001). The reduction in proptosis was statistically significant between the 2-wall and 3-wall orbital decompression groups (t=-2.82, P=0.007). Changes in BCVA, visual field, and intraocular pressure before and after surgery was greater in the 3-wall orbital decompression group compared to 2-wall orbital decompression group, although the difference was not statistically significant (all P>0.05). Change in postoperative visual acuity and visual field was significantly positively correlated with preoperative visual acuity and preoperative visual field (all P<0.001). Similarly, change in intraocular pressure and proptosis was positively correlated with preoperative intraocular pressure and preoperative protrusion (all P<0.001). Preoperative diplopia was reported in seven patients (20.0%), and two new cases (5.7%) were noted post-operation, which resolved within 3 months after surgery. Conclusions: Endoscopic endonasal decompression of the AZ area is a safe and effective surgical treatment for DON, with notable improvements in BCVA. Furthermore, three-orbital wall decompression seems to yield better outcomes in terms of eye retraction.
Subject(s)
Exophthalmos , Optic Nerve Diseases , Female , Male , Humans , Decompression, Surgical , Retrospective Studies , Lumbar Vertebrae , Optic Nerve Diseases/surgeryABSTRACT
OBJECTIVE: Exposure of oxidized low-density lipoprotein (ox-LDL) could cause dysfunction of HUVEC, thus leading to atherosclerosis development, which is a common inflammatory vascular disease. Long noncoding RNA X-inactive specific transcript (XIST) has been reported to be implicated in atherosclerosis. However, the mechanism by which this lncRNA participates in the progression of atherosclerosis is poorly defined. MATERIALS AND METHODS: HUVEC challenged by ox-LDL were used as a cellular model of atherosclerosis. Cell viability, apoptosis, LDH release, and inflammatory cytokines secretion were detected by MTT, flow cytometry, and ELISA assays. The expression levels of XIST, microRNA (miR)-30c-5p, and phosphatase and tensin homolog deleted on chromosome 10 (PTEN) were measured by quantitative Real Time-Polymerase Chain Reaction and Western blot. The target interaction between XIST and miR-30c-5p or miR-30c-5p and PTEN was validated by the Luciferase reporter assay and RNA immunoprecipitation. RESULTS: Treatment of ox-LDL induced cell apoptosis and inflammatory cytokines release in HUVEC. XIST expression was enhanced in HUVEC treated by ox-LDL, and its knockdown decreased cell apoptosis and inflammatory response in ox-LDL-treated cells. MiR-30c-5p was a target of XIST and its overexpression suppressed cell apoptosis and inflammatory response induced by ox-LDL, which was weakened by the introduction of XIST. PTEN was a target of miR-30c-5p, and its interference led to great inhibition of cell apoptosis and inflammatory response induced by ox-LDL in HUVEC, while this effect was attenuated by miR-30c-5p deficiency or XIST overexpression. CONCLUSIONS: XIST knockdown suppresses inflammatory response and apoptosis of HUVEC stimulated by ox-LDL by increasing miR-30c-5p and decreasing PTEN.
Subject(s)
Apoptosis/drug effects , Lipoproteins, LDL/toxicity , MicroRNAs/metabolism , PTEN Phosphohydrolase/metabolism , RNA, Long Noncoding/metabolism , 3' Untranslated Regions , Atherosclerosis/genetics , Atherosclerosis/metabolism , Atherosclerosis/pathology , Base Sequence , Binding Sites , Human Umbilical Vein Endothelial Cells , Humans , Interleukin-1beta/analysis , Interleukin-6/analysis , MicroRNAs/chemistry , MicroRNAs/genetics , PTEN Phosphohydrolase/chemistry , PTEN Phosphohydrolase/genetics , RNA Interference , RNA, Long Noncoding/antagonists & inhibitors , RNA, Long Noncoding/genetics , RNA, Small Interfering/metabolism , Sequence AlignmentABSTRACT
Objective: To investigate the role of body fat ratio in the evaluation of obstructive sleep apnea(OSA). Methods: A retrospective analysis was made on 174 cases (between November, 2017 and April, 2018 showed that) of sleep monitoring in the Department of Otorhinolaryngology in Peking University Third Hospital. The data included the gender, age, body fat rate, body mass index (BMI), neck circumference, and apnea-hypopnea index (AHI). The above data were analyzed by non parametric correlation analysis, receiver operating characterristic (ROC) curve analysis and multiple factor Logistic regression analysis to study the relationship between the gender,age,body fat rate,BMI,neck circumference and other indexes of the patients with AHI. Results: Nonparametric correlation analysis showed that the correlation from strong to weak to AHI among women was BMI (r=0.621, P<0.001),body fat rate (r=0.602, P<0.001), age (r=0.570, P<0.001), neck circumference (r=0.402, P=0.014), respectively. BMI (r=0.599, P<0.001), neck circumference (r=0.493, P<0.001), body fat rate (r=0.318, P<0.001), and age (r=0.256, P=0.003) among men. ROC curve analysis showed that the strong to weak index (area under curve,AUC) of the AHI>15/h among women was the body fat rate (AUC=0.884, P=0.001), BMI(AUC=0.810, P=0.008), neck circumference (AUC=0.759, P=0.027), age (AUC=0.750, P=0.033), and the male was BMI (AUC=0.765,P<0.001), neck circumference (AUC=0.720, P<0.001), age (AUC=0.634, P=0.008), and body fat rate (AUC=0.632, P=0.010), respectively. Multifactor Logistic regression analysis showed that the body fat rate (OR=1.704,95%CI=1.012-2.870) in women was an independent risk factor for AHI greater than 15/h; the age of male (OR=1. 044, 95%CI=1.005-1.085) and BMI (OR=1.285, 95%CI=1.056-1.562) were independent risk factors for AHI greater than 15/h. Conclusion: Body fat rate can be used as a new indicator for predicting the severity of OSA,especially in adult female population. In adult female moderate to severe OSA patients (AHI>15/h), compared with BMI,neck circumference and age,the body fat rate has the greatest correlation with AHI. Compared with BMI,neck circumference and age,the body fat rate has a decisive role in predicting moderate to severe OSA (AHI>15/h).
Subject(s)
Obesity/physiopathology , Sleep Apnea, Obstructive/physiopathology , Adipose Tissue/physiopathology , Adult , Body Mass Index , Female , Humans , Male , Obesity/complications , Polysomnography , Retrospective Studies , Sleep Apnea, Obstructive/etiologyABSTRACT
OBJECTIVE: Digoxin is a kind of plant-derived cardiac glycoside that is mainly used to treat heart diseases, especially in congestive heart failure or arrhythmia. However, its potentiality presented in anti-tumor remains unexplored. The purpose of this study was designed to investigate the beneficial pharmacological activity of digoxin on breast cancer cell line (MDA-MB-231, MM231). MATERIALS AND METHODS: The methyl thiazolyl tetrazolium (MTT) assay was utilized to detect the proliferation of the breast cancer MM231. The apoptotic cell numbers were determined by the flow cytometry analysis. The expressions of Bcl-2 (B-cell lymphoma-2) and Bax (Bcl2-associated X protein) were detected by Western blot analysis. RESULTS: Digoxin dose-dependently blocked the cell growth of the breast cancer MM231 through MTT assay, whereas the apoptotic numbers were significantly elevated as reflected in acridine orange staining and flow cytometry analysis. In addition, findings from Western blotting method indicated that digoxin intervention showed reduced Bcl-2 expression and elevated Bax level in MM231 cells, characterized by increased Bax/Bcl-2 ratio. CONCLUSIONS: Digoxin plays a potential anti-tumor role in breast cancer in vitro, possibly by inducing mitochondria-dependent apoptosis.
Subject(s)
Apoptosis/drug effects , Breast Neoplasms/drug therapy , Digoxin/pharmacology , Breast Neoplasms/pathology , Cell Line, Tumor , Cell Proliferation/drug effects , Female , Humans , Proto-Oncogene Proteins c-bcl-2/analysis , bcl-2-Associated X Protein/analysisABSTRACT
Objective: To evaluate the daily airborne pollen concentrations and visiting rate of patients with allergic rhinitis (AR) and their correlation during 2012-2014 in Beijing. Methods: Daily airborne pollen concentrations (55 998 numbers in total and 549 numbers in average) and its constitution from April to September each year (2012 to 2014) were compared. The number of patients with AR (44 203 in total) who visited the outpatient department of Otorhinolaryngology Head and Neck Surgery, Peking University Third Hospital between January 2012 and December 2014 was analyzed by month. Using SPSS 22.0 software, Kruskal-Wallis test was done for the comparison of visiting rate of patients with AR and airborne pollen concentrations. Correlation analysis between them was made as well. Results: χ(2) value of airborne pollen concentrations between different months in 2012 to 2014 was 110.7, 108.4 and 121.4, respectively; all P<0.01. The airborne pollen concentrations had two peaks per year, respectively: April to May, August to September. χ(2) value of visiting rate of patients with AR between different months in 2012 to 2014 was 175.0, 185.1 and 134.5, respectively; all P<0.01. Visiting rate of patients with AR showed two scattering peaks each year, respectively: April to May, August to September. The highest pollen concentration of spring (April to May) was in early and middle April. Tree pollen was the major portion in spring, which were poplar pollen, pine tree pollen, ash tree pollen, cypress tree pollen and birch trees pollen. The highest pollen concentration of autumn (August to September) was in late August and early September. Weed pollen was the major portion in summer and autumn, which were artemisia pollen, chenopodiaceae pollen and humulus japonicas pollen. The visiting rate of patients with AR showed significant correlation with airborne pollen concentrations (r value was 0.537, 0.484 and 0.566, respectively; all P<0.01). Conclusion: The visiting rate of patients with AR showed positive correlation with airborne pollen concentrations in recent three years.
Subject(s)
Allergens/analysis , Ambulatory Care Facilities/statistics & numerical data , Otolaryngology/statistics & numerical data , Pollen , Rhinitis, Allergic/epidemiology , Humans , Rhinitis, Allergic, Seasonal/epidemiology , Seasons , Software , Statistics, Nonparametric , Time Factors , TreesABSTRACT
Objective:The aim of this study is to investigate the climatic changes on spring phenology impacting on the consultation rate of patients with allergic rhinitis.Method:We analyzed the periods of full flowering stages of 15 kinds of flowers in Beijing from 2013 to 2014,and the number of patients with allergic rhinitis in the outpatient clinic of Otolaryngology Department of Peking University Third Hospital in the same time.The correlation test, analysis of variance and rank sum test were used to investigate the inherent laws and associations between the full flowering stages and the ratio of patients with allergic rhinitis.Result:â Between 2013 and 2014,the early and middle April were the full flowering stages of Platycladus orientalis,Salix matsudana, Fraxinus chinensis,Elms and Birches.The number of patients with allergic rhinitis in outpatient clinic elevated obviously in this period.â¡There were overlaps of full flowering stages among different plants.During periods in which three or more kinds of pollens in full flowering stages,the consultation rate of patients with allergic rhinitis is higher than less overlaping flower period.The difference was statistically significant(F value were -2.281,-2.964 respectively, P <0.05).Also,there were obvious positive linear correlations between the number of overlap of different pollens and the consultation rate of patients with allergic rhinitis in in 2013 and 2014(r=0.957, 0.964 respectively).Conclusion:During the overlaping flower periods,there was a higher consultation rate of patients with allergic rhinitis.In prevention and treatment of allergic rhinitis,the overlapping effect of full flowering stages of different pollens should be taken into account.Upgrade the alarm level will reduce the incidence of AR and the degree of allergic rhinitis episodes.