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Objective:To explore the inhibitory effect of Shiquandabu decoction on the spontaneous len tumor of the SV40 T antigen transgenic (TG) mice,and to clarify its molecular mechanism.Methods:The SV40 T antigen TG mice were randomly divided into control group (n=39) and drug treatment group (n=25).The mice in control group were fed normally,while the mice in drug treatment group were fed with Shiquandabu decoction at the 3rd week after birth,the survival time of mice was recorded.Three mice in control group and drug treatment group were randomly chosen to collect the blood from the tail vein and the amino acid levels were measured respectively 8 and 15 weeks after Shiquandabu decoction administration.Then the mice were sacrificed and the liver tissue wascollected.Gene chip hybridization was used to detect the differences in the expressions of ribosomal function related genes in liver tissue of the mice in two groups and the related signal pathway was explored.Results:The survival analysis demonstrated that the survival rate of TG mice in drug treatment was higher than that in control group (P＜0.05).Compared with control group,the serum levels of alanine,valine,leucine,isoleucine,threonine,methionine,proline,tyrosine,lysine,sarcosine,citrulline,ornithine and hydroxylysine of the mice in drug treatment group 8 weeks after administration of Shiquandabu decoction were increased (P＜0.05);and the serum levels of cystathionine,taurine,methylhistidine,anserine and ethanolamine were decreased (P＜0.05).Fifteen weeks after administration,compare with control group,the serum levels of threonine and citrulline of the mice in drug teeatment group were increased (P＜0.05),but the serum levels of other amino acids had no significant difference (P＞ 0.05).The canonical analysis showed that thirteen genes involved in ribosomal function from 9 083 genes in liver tissue in drug treatment group had the changes compared with control group (P＜ 0.05).Conclusion:Shiquandabu decoction can effectively prolong the lifetime of the TG mice by improving the levels of serum amino acids and promoting the liver ribosomal protein synthesis.
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Objective To investigate the clinicopathological and prognostic significance of lysosome-associated membrane protein-2a (LAMP2a) expression in the development of colorectal cancer.Methods Immunohistochemistry was performed to detect the LAMP2a expression in colorectal cancer,normal mucosa,and adenoma tissues.Results LAMP2a expression in colorectal cancer and adenoma was higher than that in normal mucosa (P ＜ 0.05).Further,LAMP2a expression in colorectal cancer was negatively correlated with TNM staging and lymph node metastasis (P ＜ 0.05);however,the invasion and liver metastasis,depth of invasion and differentiation had no correlation with the lymph node (P ＞ 0.05).Kaplan-Meier analysis showed that LAMP2a expression in colorectal cancer is not significantly correlated with good prognosis (P ＞ 0.05).Univariate survival analysis showed that lymphatic invasion,vascular invasion,lymph node metastasis,TNM stage,liver metastasis,distant metastasis,and differentiation were correlated with poor prognosis of colorectal cancer (P ＜ 0.05).The Cox proportional hazards regression model showed that the depth of invasion and distant metastasis were important factors affecting the survival period of patients with colorectal cancer (P ＜ 0.05).Conclusion LAMP2a may play a role in the carcinogenesis and progression of colorectal cancer,and may be used as a molecular marker for the biological behavior of colorectal cancer.
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Objective To investigate the effects of inhibitor of growth 5 (ING5) gene on the proliferation, apoptosis, migration and cell cycle of human breast cancer Bcap-37 cells.Methods The eukaryotic ING5-expressing plasmid and GFP-empty plasmid were steadily transfected in Bcap-37 cells, the expression of green fluorescent protein was measured with fluorescence microscopy, and the high expression of ING5 was measured by real time-PCR. Bcap-37-ING5 cells served as the experimental group, Bcap-37-GFP cells as the mock group and Bcap-37 as the control group. The effects of ING5 on the proliferation were detected by MTT, the cell cycle and apoptosis were detected by Flow cytometry, and the cell migration was detected by cell wound scratch assay and Transwell experiment.Results Bcap-37 cell lines steadily expressing ING5 protein with GFP-tag were acquired by stable transfection. ING5 over-expression inhibited the proliferation and led to G2 arrest of Bcap-37 cells, increased cells apoptosis and decreased the cell migration ability (P<0.05).Conclusion ING5 over-expression may have reverse effect for malignant phenotype of breast cancer cells, and may be employed to indicate the biomarker of prognosis of breast cancer patients and regarded as a target of gene therapy.
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Objective To explore the effects and molecular mechanisms of suberoylanilide hydroxamic acid (SAHA) on ovarian carcinoma. Methods (1)Two groups of ovarian carcinoma cell lines (SKOV3 and SKOV3/DDP, HO8910 and HO8910-PM) were exposed to SAHA (1, 3, 5 and 7μmol/L SAHA,group 1-group 4). CCK-8 method was employed to eval-uate the inhibitory effects of SAHA.(2)Ovarian cancer cell lines treated with SAHA (2 or 5μmol/L SAHA) were used as 1 and 2 groups. Flow cytometry was performed following staining with Annexin V-FITC and PI for cell cycle and apoptosis.(3) Reverse transcription polymerase chain reaction (RT-PCR) and Western blot assay were used to assess the mRNA and pro-tein expression levels of phenotypic correlation factor. Results (1)After 48 h of SAHA treatment,the OD value of SKOV3, SKOV3/DDP,and HO8910 showed a trend of gradually reduce (P<0.05).(2)The apoptotic rates were significantly higher in SAHA 1 and SAHA 2 groups than those of control group (P<0.05). Compared with control group, after 48 h of SAHA treat-ment,S phase and G2/M phase of SKOV3 and SKOV3/DDP cells increased;G0/G1 phase of HO8910 and HO8910-PM cells increased in SAHA 1 and 2 groups (P<0.05).(3)The expression levels of CyclinB1 and Cdc2 (p34) mRNA were significant-ly lower in SAHA 1 and 2 groups than those of control group,while the expression levels of Caspase-3,p21 and p53 mRNA expression were significantly higher in SAHA 1 and 2 groups than those of control group. Furthermore,the expression of Ac-Histone H3,Ac-Histone H4,p53 protein were markedly improved,and CyclinB1,Cdc2(p34) protein decreased in SAHA 1-4 groups. Conclusion SAHA may suppress cell growth, induce apoptosis and cause cycle arrest in ovarian carcinoma cells by promoting histone acetylation or modulating their phenotype-related proteins of Caspase-3, p53, CyclinB1 and Cdc2(p34).
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Objective To study the effect of ING5 gene on growth inhibition of gastric cancer. Methods ING5 expressing plasmid was transfect?ed into SGC?7901 cells. The cell viability was assessed by CCK?8,cell apoptosis and cycle was detected by flow cytometry analysis,the migration and invasion was evaluated by scratch and transwell,the expressions of mRNA and protein were determined by real?time quantitative PCR and West?ern blot respectively. Nude mice were implanted subcutaneously with SGC?7901 cells and tumor size and related protein were analyzed. Results After transfection of pEGFP?N1?ING5,the proliferation of gastric cancer SGC?7901 cells was significantly inhibited(P<0.05),the apoptosis was decreased(P<0.05),the percentage of G1 phase was increased and G2 phase was decreased(P<0.05),the ability of migration and invasion was reduced(P<0.05),the expression of NF?κB,PI3K,p?Akt1/2/3,Bcl?2,XIAP,β?catenin,c?myc mRNA,and protein levels were significantly in?creased(P<0.05),and the expression of Akt1/2/3、MMP9 mRNA and protein levels were significantly decreased(P<0.05). Conclusion The ING5gene inhibits the SGC?7901 cell proliferation,induces G1 arrest,inhibits the migration and invasion,and effectively regulates the related genes and proteins about cell proliferation,cell cycle and adhesion,but reduces apoptosis. ING5 can inhibit the evolution and development of gastric can?cer.
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Objective Rho signaling component α-catulin,is a cytoskeletal linker protein and plays an important role in apoptotic and senescence resistance,cytoskeletal reorganization,mobility,invasion and epithelial to mesenchymal transition(EMt)of cancer cells. Methods Here,we ex-amined α-catulin expression in squamous epithelium,dysplasia and cancer of head and neck on tissue microarrays by immunostaining. Its expres-sion was compared with clinicopathological parameters and survival rate of cancers. Results It was found that α-catulin expression level was signifi-cantly higher in primary cancers than that in normal squamous epithelium and dysplasia(P 0.05). Cox′s proportional hazard model indicated that distant metastasis and tNM staging were independent prognostic factors for overall survival of the patients with head and neck squamous carcinoma(HNSCC,P < 0.05). Conclusion these findings suggested that up-regulated expression of α-catulin protein may play an important role in the pathogenesis of HNSCC,which might be employed as a potential marker for tumorgenesis of HNSCCs.
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Objective to explore the role of Beclin 1 in gastric carcinogenesis and subsequent progression. Methods MkN28 cells were trans-fected with Beclin 1-expressing plasmid,and then the proliferation and cell cycle was measured by CCk-8 and PI staining. Beclin 1 expression was examined by immunohistochemistry and in situ hybridization on tissue microarrays containing gastric cancers,adjacent non-neoplastic mucosa,and metastatic lymph node. the correlation with the tumorgenesis,clinicopathological and prognostic parameters was analyzed. Results Beclin 1 overex-pression resulted in G2 arrest of MkN28 cells and reduced the proliferation. Beclin 1 mRNA was highly expressed in gastric cancer than matched mu-cosa by ISH(P < 0.05). Beclin 1 was highly expressed in male than female patients with gastric cancer(P < 0.05). the elder patients with gastric cancer had higher Beclin 1 expression than younger ones(P < 0.05). the diffuse-type carcinomas showed less Beclin 1 expression than intestinal and mixed type ones(P < 0.05). kaplan-Meier analysis indicated that Beclin 1 expression was positively correlated to favorable prognosis of the can-cer patients(P < 0.05). Conclusion Beclin 1 expression is closely linked to pathogenesis,metastasis and differentiation of gastric cancer. Beclin 1 might be employed to indicate the favorable prognosis of gastric cancer patients and regarded as a target of gene therapy.
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Objective To clarify the clinicopathological significance and the reversing effects of BTG3 expression on the aggressive phenotype in gastric cancer. Methods BTG3 expression was detected in gastric cancer tissues by on tissue microarray and immunostaining. BTG3?expressing plasmid was transfected into MKN28 and MGC803 cells,the proliferation,cell cycle,differentiation and autophagy were analyzed by CCK?8,PI staining,alkaline phosphatase activity and GFP?LC?3B transfection,respectively. Results BTG3 overexpression inhibited cell proliferation,in?duced S/G2 arrest,differentiation and autophagy in both cells(P<0.05). BTG3 expression was decreased in gastric cancer in comparison with the adjacent mucosa(P<0.05),and positively correlated with venous invasion and dedifferentiation of the cancers(P<0.05). Conclusion BTG3 ex?pression contributes to gastric carcinogenesis and subsequent progression. BTG3 overexpression can reverse the aggressive phenotypes,which could be employed as a potential target for gene therapy of gastric cancer.
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AIM:To investigate the effects of 17-AAG on apoptosis and cell cycle of HCT-15 cells and to clar-ify the related mechanisms .METHODS: MTT method was employed to evaluate the inhibitory effects of 17-AAG with Aifferent time and different doses on the proliferation of HCT-15 cells.The cells were stained with Annexin V-FITC/propid-iumiodide and measured by flow cytometry .The expression of STAT3, cyclin D1, Cyt C, caspase 9 and caspase 3 at mR-NA and protein levels was determined by RT-PCR and Western blotting .RESULTS:Treatment with 17-AAG at concentra-tion of 1.25~20 mg/L for 24 h and 48 h significantly inhibited the activity of HCT-15 cells at both time-and concentra-tion-dependent manners .Treatment with 17-AAG at concentrations of 0.425, 0.85 and 1.7 mg/L for 48 h significantly in-duced apoptosis and cell cycle arrest of HCT-15 cells.The exposure of 17-AAG at concentrations of 0.425, 0.85 and 1.7 mg/L for 48 h to the HCT-15 cells significantly down-regulated the expression of STAT 3 and cyclin D1 at mRNA and pro-tein levels, but up-regulated Cyt C, caspase 9 and caspase 3 mRNA and protein in a concentration-dependent manner . CONCLUSION:17-AAG inhibits the cell activity , induces apoptosis and G 1 arrest by down-regulating the expression of cyclin D1, and promoting the mitochondria apoptosis through STAT 3 pathway.
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Objective To construct and confirm the JC virus(JCV) T antigen expression plasmid using mouse keratin 19 (K19) promoter specific for the gastric epithelial cells.Methods The Ndel site was mutated by FCR with Bell insertion at both sides.The DNA fragment digested by Bcl Ⅰ was ligated with the plasmid containing K19 promoter via Bam Ⅰ site.The DNA sequence was confirmed by restriction enzyme digestion and direct DNA sequencing.Cytokeratin 19 protein was examined to screen gastric carcinoma cell for transfection of K19-JCV T antigen expression plasmid by immunohistochemistry.The Western blot was employed to detect the JCV T antigen expression in the gastric carcinoma transfectant.Results K19-JCV T antigen expressing plasmid was successfully constructed.The ACS strongly expressed cytokeratin 19 protein and was selected for the transfection of K19-JCV T antigen expressing plasmid.JCV T antigen was positively expressed in the AGS transfectant.Conclusion The synonymous mutation and compatible ligation are useful in the plasmid construction.The methy lation of restriction enzyme should be considered.It is meaning for the transgenic animal model of gastric carcinoma to successfully construct the JC virus T antigen expression plasmid in gastric mucosa.
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<p><b>BACKGROUND</b>It has been known that Kai1 is one of transmembrane 4 superfamily regulating cell proliferation, adhesion and mobility and its down-regulated expression is closely correlated with progression and prognosis of tumor. Fas ligand (FasL) is one of tumor necrosis factor/nerve growth factor family activating caspase-3, a key proteinase in cell apoptosis and leading immune escape in tumor cells. The aim of this study is to investigate the expression of Kai1 and FasL in non-small cell lung cancer (NSCLC) and to explore their roles and relationship in carcinogenesis and progression of NSCLC.</p><p><b>METHODS</b>Kai1 and FasL expressions were examined in 79 NSCLC tissues and their adjacent normal lung tissues by SP immunohistochemistry. Their expressions were compared with clinicopathological features of NSCLC. The relationship between Kai1 and FasL expressions was also analyzed in NSCLC.</p><p><b>RESULTS</b>Kai1 expression in NSCLC tissue was remarkably lower than that in their adjacent tissue (55.7% vs 82.6%, P < 0.05). However, it was the converse for FasL (73.4% vs 52.2%, P < 0.05). Kai1 expression was not correlated with age and gender of NSCLC patients, tumor location or histological classification (P > 0.05), but negatively with lymph node metastasis and clinicopathological staging and positively with differentiation degree (P < 0.05). FasL expression was not correlated with age and gender of the patients, tumor location or histological classification (P > 0.05), but positively with lymph node metastasis and negatively with differentiation degree (P < 0.05). Kai1 expression was closely correlated with FasL expression in NSCLC (P < 0.05).</p><p><b>CONCLUSIONS</b>Down-regulation of Kai1 expression and up-regulation of FasL may play important roles in carcinogenesis and progression of NSCLC. Kai1 and FasL could be considered as molecular markers to reflect pathobiological behavior of NSCLC. Additionally, close correlation of FasL expression with Kai1 expression in NSCLC provides a novel insight into the regulatory effects of FasL expression on Kai1 expression.</p>
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<p><b>OBJECTIVE</b>To investigate the expression of PTEN and Caspase-3 in malignant lymphoma of the stomach and explore their role in progression of primary gastric malignant lymphoma.</p><p><b>METHODS</b>Formalin-fixed paraffin embedded tissues from 56 cases of primary gastric malignant lymphoma and their adjacent non-tumor mucosa were evaluated for PTEN and Caspase-3 protein expression by streptavidin-biotin-complex (SABC) immunohistochemistry. Their expression was compared with clinical tumor parameters with the relationship between PTEN and Caspase-3 expression concerned as well.</p><p><b>RESULTS</b>The positive rate of PTEN expression in primary gastric lymphomas (50.0%, 28/56) was significantly lower than that in adjacent non-tumor gastric mucosa (96.4%, 27/28) (P < 0.05). Meanwhile, 43 of 56 (76.8%) gastric lymphomas indicated Caspase-3 expression, less than that in adjacent non-tumor mucosa (93.5%, 29/31) (P < 0.05). The expression of PTEN was negatively correlated with invasion and lymph node metastasis of gastric lymphoma (P < 0.05), while the Caspase-3 expression was negatively associated with the latter one (P < 0.05). Additionally, the PTEN expression was positively correlated with Caspase-3 expression in the primary gastric malignant lymphoma (P < 0.05).</p><p><b>CONCLUSIONS</b>The down-regulated expression of PTEN and Caspase-3 played an important role in progression of primary malignant gastric lymphoma. PTEN, as a molecular marker of pathobiological behaviors of tumor, contributes to tumor progression by increasing cell mobility and angiogenesis, as well as decreasing cell adhesion and apoptosis.</p>
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Adult , Aged , Female , Humans , Male , Middle Aged , Biomarkers, Tumor , Metabolism , Caspase 3 , Caspases , Metabolism , Down-Regulation , Gastric Mucosa , Lymphatic Metastasis , Lymphoma , Pathology , Neoplasm Invasiveness , PTEN Phosphohydrolase , Phosphoric Monoester Hydrolases , Metabolism , Stomach Neoplasms , Pathology , Tumor Suppressor Proteins , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the mutation and expression of tumor suppressor gene-PTEN mRNA and explore their roles in tumorigenesis and progression of ovarian cancer.</p><p><b>METHODS</b>Mutated exon 5 of PTEN gene was examined in normal ovary (n=5), ovarian cyst (n=5), ovarian borderline tumor (n=9), epithelial ovarian cancer (n=60), and ovarian cancer cell line (n=1) by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). mRNA expression of PTEN gene was evaluated in corresponding tissues and cell line by reverse transcription polymerase chain reaction (RT-PCR). The mutation and mRNA expression of PTEN gene were compared with clinicopathological features of ovarian cancer.</p><p><b>RESULTS</b>Mutated exon 5 of PTEN gene was detected only in 5 (7.1%) cases of epithelial ovarian cancer. mRNA expression level of PTEN gene in ovarian borderline tumor or ovarian cancer was lower than that in normal ovary or ovarian cyst (P < 0.05). The level of PTEN gene mRNA expression was negatively correlated with clinicopathological staging of ovarian cancer, whereas positively correlated with histological differentiation (P < 0.05). mRNA expression level of PTEN gene in ovarian endometrioid cancer was significantly lower than that in ovarian serous or mucinous cancer (P < 0.05).</p><p><b>CONCLUSIONS</b>Mutation of PTEN gene occurs in ovarian cancer. Down-regulated expression of PTEN is probably an important molecular event in tumorigenesis of ovarian cancer. Abnormal expression of PTEN gene is involved in progression of ovarian cancer. Reduced expression of PTEN gene is closely associated with tumorigenesis and pathobiological behaviors of ovarian endometrioid cancer.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Middle Aged , Carcinoma, Endometrioid , Genetics , Down-Regulation , Exons , Gene Expression Regulation, Neoplastic , Genes, Tumor Suppressor , Mutation , Ovarian Neoplasms , Genetics , PTEN Phosphohydrolase , Phosphoric Monoester Hydrolases , Genetics , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , RNA, Messenger , Genetics , Tumor Suppressor Proteins , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To clarify the role of midkine (MK) in vulvar carcinogenesis though examination of its expression in vulvar lesions including vulvar condyloma acuminata (VCA), vulvar intraepithelial neoplasia (VIN) and vulvar squamous cell carcinomas (VSCC), and to analyze the relationship between MK expression and human papilloma virus (HPV) infection.</p><p><b>METHODS</b>Thirty VSCC, 15 VIN and 10 VCA patients were studied by streptavidin-biotin-immunoperoxidase method. MK expression was compared with clinicopathologic features of vulvar tumors.</p><p><b>RESULTS</b>MK was expressed in 26 of 30 VSCC (87%), 3 of 5 VIN III and all VCA samples, whereas no MK expression was detected in the VIN I-II samples or in normal epithelium. The difference of MK expression between VIN III and VSCC was statistically significant (P < 0.05). MK was more intensely expressed in differentiated-type (well differentiated and moderately differentiated) VSCC than in undifferentiated-type (poorly differentiated) VSCC. There was no statistically significant correlation between MK expression and clinical stage, lymph node metastasis and HPV infection in VSCC. MK expression were observed in all HPV-positive specimens including 2 VSCC, 1 VIN III and all VCA.</p><p><b>CONCLUSIONS</b>MK gene expression may be a late event in vulvar squamous cell malignant transformation, and may be associated with vulvar tumor cell differentiation. HPV-positive vulvar tumors expressed MK protein.</p>