ABSTRACT
BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is increasingly recognized as a chronic, progressive and fatal lung disease with an unknown etiology. Current studies focus on revealing the genetic factors in the risk of IPF, making the integrative analysis of genetic variations and transcriptomic alterations of substantial value. AIM: This study aimed to improve the understanding of the molecular basis of IPF through an integrative analysis of whole-exome sequencing (WES), bulk RNA sequencing (RNA-seq) and single-cell RNA sequencing (scRNA-seq) data. METHODS: WES is a powerful tool for studying the genetic basis of IPF, allowing for the identification of genetic variants that may be associated with the development of the disease. RNA-seq data provide a comprehensive view of the transcriptional changes in IPF patients, while scRNA-seq data offer a more granule view of cell-type-specific alterations. RESULTS: In this study, we identified a comprehensive mutational landscape of recurrent genomic and transcriptomic variations, including single-nucleotide polymorphisms, CNVs and differentially expressed genes, in IPF populations, which may play a significant role in the development and progression of IPF. CONCLUSIONS: Our study provided valuable insights into the genetic and transcriptomic variations associated with IPF, revealing changes in gene expression that may contribute to disease development and progression. These findings highlight the importance of an integrative approach to understanding the molecular mechanisms underlying IPF and may pave the way for identifying potential therapeutic targets.
Subject(s)
Idiopathic Pulmonary Fibrosis , Humans , Idiopathic Pulmonary Fibrosis/genetics , Gene Expression Profiling , MutationABSTRACT
Objective: To investigate the regulatory mechanisms of piwi-interacting RNA (piRNA) in bisphenol A (BPA)-induced prostate cancer cell invasion and migration. Methods: The Cancer Genome Atlas (TCGA) data was used to analyze and screen for piRNAs with significantly increased expression in prostate cancer tissues. PC-3 cells were treated with different concentrations of BPA for 12, 24, and 48 h, respectively, and the 20% inhibitory concentration (IC20) was measured using a CCK-8 assay. The expression levels of piRNAs before and after BPA treatment were determined by reverse transcription-quantitative PCR. Target genes regulated by BPA and associated with prostate cancer were screened in the Comparative Toxicogenomics Database (CTD). Dual-luciferase reporter gene assay was performed to verify the relationship between piRNA and target genes, and the expression change of the piRNA target gene was detected by Western blotting. Cell migration and invasion assays were used to determine the effects of piRNA on the malignant phenotype of prostate cancer cells. Results: After treatment of PC-3 cells with 160 µmol/L BPA, the expression of piR-sno48 was most significantly increased (P<0.05). Transfection of piR-sno48 antagomir resulted in decreased expression of endogenous piR-sno48 and a significant increase in the expression of its target gene GSTP1 (P<0.05). However, the expression of GSTP1 did not change significantly in BPA-treated PC-3 cells after transfection with piR-sno48 antagomir (P>0.05). The dual-luciferase reporter gene confirmed that piR-sno48 inhibited the expression of GSTP1 by forming an inversely complementary sequence with the 3'-UTR of GSTP1. The Transwell assay results showed that treatment with BPA significantly increased the invasion and migration ability of prostate cancer cells (P<0.01), whereas piR-sno48 antagonists significantly inhibited the effects above (P<0.01). Conclusion: BPA promotes the invasion and migration of prostate cancer cells by upregulating the expression of piR-sno48 and suppressing the expression of GSTP1. Interfering with the expression of endogenous piR-sno48 may inhibit the malignant phenotype of prostate cancer cells caused by BPA.
Subject(s)
Prostate , Prostatic Neoplasms , Male , Humans , Piwi-Interacting RNA , Antagomirs , Prostatic Neoplasms/geneticsABSTRACT
Infant intestinal development is immature and, thus, is vulnerable to bacterial and viral infections, which damage intestinal development and even induce acute enteritis. Numerous studies have investigated that lactoferrin (LF) has protective effects on the intestine and may play a role in preventing intestinal inflammation in infants. Lactoferrin is divided into 2 types, namely apo-LF and holo-LF, depending on the degree of iron saturation, which may affect its bioactivities. However, the role of LF iron saturation in protecting infant intestinal inflammation has not been clearly clarified. Therefore, in this study, young mice models with intestinal damage induced by lipopolysaccharides (LPS) in vivo and primary intestinal epithelial cells in vitro were constructed to enteritis injury in infants for investigation. The apo-LF and holo-LF were subsequently applied to the mouse models to investigate and compare their levels of protection in the intestinal inflammatory injury, as well as to identify which LF was most active. Moreover, the specific mechanism of the LF with optimal iron saturation was further investigated through Western blot assay. Results demonstrated that disease activity index, shortened length of colon tissue, and histopathological score were significantly decreased in the apo-LF group compared with those of the LPS group and the holo-LF group. In the apo-LF group, the concentration of LPS in the intestinal tract and the number of gram-negative bacteria colonies decreased significantly and the expression levels of proinflammatory factors in the colon tissue were downregulated, in comparison with those in the LPS group. The findings of this study thus verify that apo-LF can significantly alleviate enteritis injury caused by LPS, through regulating the PPAR-γ/PFKFB3/NF-κB inflammatory pathway.
Subject(s)
Enteritis , Iron , Lactoferrin , Animals , Enteritis/prevention & control , Enteritis/veterinary , Inflammation/veterinary , Iron/metabolism , Lactoferrin/pharmacology , Lipopolysaccharides , Mice , Recombinant Proteins/pharmacologyABSTRACT
The development of high-performance ultraelastic metals with superb strength, a large elastic strain limit and temperature-insensitive elastic modulus (Elinvar effect) are important for various industrial applications, from actuators and medical devices to high-precision instruments1,2. The elastic strain limit of bulk crystalline metals is usually less than 1 per cent, owing to dislocation easy gliding. Shape memory alloys3-including gum metals4,5 and strain glass alloys6,7-may attain an elastic strain limit up to several per cent, although this is the result of pseudo-elasticity and is accompanied by large energy dissipation3. Recently, chemically complex alloys, such as 'high-entropy' alloys8, have attracted tremendous research interest owing to their promising properties9-15. In this work we report on a chemically complex alloy with a large atomic size misfit usually unaffordable in conventional alloys. The alloy exhibits a high elastic strain limit (approximately 2 per cent) and a very low internal friction (less than 2 × 10-4) at room temperature. More interestingly, this alloy exhibits an extraordinary Elinvar effect, maintaining near-constant elastic modulus between room temperature and 627 degrees Celsius (900 kelvin), which is, to our knowledge, unmatched by the existing alloys hitherto reported.
ABSTRACT
Objective: Diet balance index (DBI_16) was used to evaluate the dietary status of smoking adults in Tianjin, and the relationship between DBI_16 and serum uric acid was analyzed. Methods: A total of 1 478 inhabitants aged 18 and above were enrolled. The dietary status was obtained with a 3-day dietary recall and condiment weighing method. And their fasting venous blood was collected to detect uric acid. Food intake and DBI_16 scores of smokers and non-smokers were described, and the relationship among smoking, diet quality distance (DQD-DBI), and serum uric acid was analyzed. Results: The intake of vegetables, fruits, and milk of smokers in Tianjin was lower than while the salt, oil, cereals, and aquatic products were higher than that of non-smokers. The DBI_16 scores of vegetables and fruits, food types, milk, and beans of smokers were lower than those of non-smokers in Tianjin, and the scores of pure energy foods and condiments were higher than those of the non-smokers. The DQD-DBI, high bound score and low bound score of smokers in Tianjin were 42.0, 14.0, and 29.0 respectively, which were all higher than those of non-smokers. The main problems appeared as moderately inadequate intake (accounting for 67.0%), low, excessive intake (accounting for 70.9%), and moderate imbalance of intake (accounting for 67.2%). The serum uric acid of smokers was higher than in the non-smokers, and there the same result appeared under the conditions of "not suitable" in DQD-DBI. Conclusions: In Tianjin, the dietary imbalance was more evident in smokers than the non-smokers, and the serum uric acid was significantly higher than the non-smokers. Strategies as strengthening the nutrition education and intervention targeted for smokers were in urgent need.
Subject(s)
Diet , Uric Acid , Adult , Diet Surveys , Humans , Smoking , VegetablesABSTRACT
Objective: To understand the influence of birth weight on the risk of chronic diseases, such as hypertension and diabetes, and the relationship between birth weight and serum uric acid in adulthood. Methods: According to the Chinese Adult Chronic Disease and Nutrition Surveillance Program, a total of 1 131 residents aged 18 years and above were enrolled from Hexi, Nankai, Hongqiao, Wuqing, Jinnan, Baodi and Jizhou districts of Tianjin. The data of birth weight and blood pressure of the residents were collected, and fasting venous blood samples were collected from them to detect uric acid, blood glucose and blood lipids levels. The distribution of birth weight of the surveyed population was described, and the relationship between birth weight and chronic diseases, such as hypertension and diabetes, and the blood uric acid level in adulthood were analyzed. Results: The average birth weight of the surveyed population in Tianjin was 3.37 kg, which was higher in males than in females. No matter overweight/obesity, hypertension or diabetes, the prevalence rate of normal birth weight was the lowest in adulthood. After univariate logistic regression analysis and adjusting for age, sex, smoking and drinking status, it was found that compared with normal birth weight, low birth weight had a stronger correlation with diabetes (OR=2.91,95%CI:1.46-5.76) and dyslipidemia (OR=1.79,95%CI:1.01-3.19) in adulthood. Macrosomia was strongly associated with overweight/obesity in adulthood (OR=1.47, 95%CI:1.08-2.01). There was no significant difference in serum uric acid level among the residents with different birth weights. Conclusions: The low birth weight of the surveyed population in Tianjin was closely related to the risk of diabetes and dyslipidemia in adulthood, and the macrosomia was closely related to the risk of overweight/obesity in adulthood.
Subject(s)
Fetal Macrosomia , Uric Acid , Adult , Birth Weight , Body Mass Index , Chronic Disease , Female , Humans , Male , Pregnancy , Risk FactorsABSTRACT
This study tested the ability of lactoferrin to modulate pulmonary inflammation. To construct in vitro and in vivo inflammatory lung models, cells from the human lung adenocarcinoma cell line (A549) were exposed to lipopolysaccharide (LPS, 1 µg/mL), and mice (CD-1) were intratracheally administered LPS [10 mg/kg of body weight (BW), tracheal lumen injection], respectively. The A549 cells were preincubated with lactoferrin (10 mg/mL), and the mice were intraperitoneally injected with lactoferrin (100 mg/kg of BW), followed by LPS treatment. The concentrations of proinflammatory cytokines (IL-1ß and TNF-α) in culture medium of A549 cells and in bronchoalveolar lavage fluid of the mice were determined using enzyme-linked immunosorbent assays. The toll-like receptor 4-related pathway (TLR4/MyD88/IRAK1/TRAF6/NFκB) was determined at gene and protein expression levels in A549 cells and mouse lung tissue. Results showed that LPS treatment significantly elevated the concentrations of IL-1ß and TNF-α in the A549 cell culture medium and in bronchoalveolar lavage fluid of the mice; it also elevated both the mRNA and protein expressions of TLR4 and the TLR4 downstream factors in A549 cells and mouse lung tissue. Nevertheless, lactoferrin apparently depressed the releases of IL-1ß and TNF-α from A549 cells and lung tissues stimulated by LPS, and significantly suppressed the TLR4 signaling pathway. Lactoferrin also promoted the enhancement of miR-146a expression in A549 cells and mouse lung tissue. Moreover, 100°C heating for 3 min caused total loss of the previously listed bioactivity of lactoferrin. Collectively, we proved that lactoferrin intervened in LPS-induced inflammation in the pulmonary cell model and in the mouse model, through inhibiting the TLR4-related pathway.
Subject(s)
Pneumonia , Rodent Diseases , Animals , Lactoferrin , Lipopolysaccharides , Lung , Mice , NF-kappa B/metabolism , Pneumonia/veterinary , Toll-Like Receptor 4/metabolismABSTRACT
OBJECTIVE: The aim of this study was to explore the relationships between ADAMTS-13 gene polymorphisms and hypertension-induced atrial fibrillation (AF). PATIENTS AND METHODS: A total of 200 hypertensive patients without AF (hypertension group) and 200 hypertensive patients with AF (AF group) treated in our hospital were enrolled. Then, peripheral blood was drawn from these subjects enrolled, and the genomic deoxyribonucleic acids (DNAs) were extracted for analysis of ADAMTS-13 gene polymorphism. Next, Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) was employed to determine the expression of ADAMTS-13 gene, and the correlations of ADAMTS-13 gene polymorphism with ADAMTS-13 gene expression and clinical indicators were analyzed. RESULTS: Results revealed that there was a difference in the distribution of alleles of ADSMTS-13 rs3094374 (p=0.046) and rs34054981 (p=0.039) between AF group and hypertension group. The frequency of T allele of the locus rs3094374 and that of the locus rs34054981 in ADSMTS-13 gene was higher in AF group than that in hypertension group. The distribution of genotypes of ADSMTS-13 rs28503257 (p=0.047) and rs34054981 (p=0.013) in AF group were different from those in hypertension group, and AF group had lower frequency of GA genotype of ADAMTS-13 rs28503257 and higher frequency of CT genotype of ADAMTS-13 rs34054981 than hypertension group. Besides, a difference was found in the distribution of ADSMTS-13 rs3094374 between AF group and hypertension group in recessive model (p=0.043), and the frequency of TC + CC was higher in the recessive model. Moreover, the distribution of the haplotypes CAT (p=0.012) and CGT (p=0.031) in ADAMTS-13 gene showed a difference between AF group and hypertension group. The linkage disequilibrium of the loci rs3094374 and rs28503257 in ADAMTS-13 gene was relatively great (D'=0.293). In addition, the polymorphism of the locus rs34054981 in ADAMTS-13 gene had an association with ADAMTS-13 gene expression (p<0.05). The expression of ADAMTS-13 gene was lower in patients carrying genotype TT in AF group. Furthermore, the ADAMTS-13 rs3094374 polymorphism was related to international normalized ratio (INR) (p=0.034), and the ADAMTS-13 rs28503257 polymorphism was correlated with the levels of brain natriuretic peptide (BNP) (p=0.047) and D-dimer (p=0.033). CONCLUSIONS: ADAMTS-13 gene polymorphism is correlated with the susceptibility and procession of hypertension-induced AF.
Subject(s)
ADAMTS13 Protein/genetics , Atrial Fibrillation/genetics , Hypertension/genetics , Polymorphism, Single Nucleotide/genetics , ADAMTS13 Protein/blood , Atrial Fibrillation/blood , Atrial Fibrillation/diagnosis , Female , Humans , Hypertension/complications , Hypertension/diagnosis , Male , Middle AgedABSTRACT
Objective: To investigate the relationship between preoperative evaluation, surgery and prognosis of microvascular decompression (MVD) for the treatment of hemifacial spasm (HFS). Methods: The clinical data of 128 HFS patients treated with MVD in the department of neurosurgery of Taizhou Hospital of Zhejiang Province were retrospectively analyzed. According to the SMC grading system, the patients were divided into general spasm group and severe spasm group, and the clinical characteristics, offending vessel, prognosis and surgical complications of the two groups were compared. Results: In the general spasm group,the age at MVD was (48.6±10.6) years, the disease duration was (4.2±3.3) years;while in the severe spasm group,the age at MVD was (51.8±9.9) years, the disease duration was (8.1±4.5) years;the differences of age and disease duration between the two groups were statistically significant (P<0.05).In the general spasm group, there were 41 cases in which the offending vessel were AICA, 21 cases were PICA, 1 case was VA, 63 cases were single offending vessel, and 7 cases were multiple offending vessels. In the severe spasm group, there were 29 cases in which the offending vessel were AICA, 13 cases were PICA, 2 cases were VA, the total of 44 cases were single offending vessel and 14 cases were multiple offending vessels.There was a significant difference in the proportion of multiple offending vessels in the two groups, and the difference was statistically significant (P<0.05).Patients in the two groups were followed up for 12 to 32 months after surgery, and the difference in effective rate and recurrence rate was not statistically significant (P>0.05).Some kinds of postoperative complications were different between the two groups, the incidence of postoperative delayed facial paralysis was statistically significant (P<0.05), and the other complications were not statistically significant (P>0.05). Conclusion: Compared with the general spasm group, the patients in the severe spasm group were older, with longer disease duration, higher probability of multiple offending vessels and higher incidence of postoperative delayed facial paralysis. Therefore, preoperative SMC grading is helpful for the evaluation and prediction of intraoperative and postoperative conditions, which is worthy of wide clinical application.
Subject(s)
Facial Paralysis , Hemifacial Spasm , Microvascular Decompression Surgery , Adult , Humans , Middle Aged , Prognosis , Retrospective Studies , Treatment OutcomeABSTRACT
Objective: To explore the effects of sodium arsenite (NaAsO(2)) exposure on the activation and extracellular matrix secretion of human hepatic stellate cells, and to provide a theoretical basis for the mechanism study of arsenic induced hepatic fibrosis. Methods: Different doses of NaAsO(2) (0.0, 0.1, 1.0, 10.0, 50.0, 100.0 µmol/L) were exposed to human hepatic stellate cell line (Lx-2) for 24, 48 and 72 huors. CCK-8 assay was used to measure cell viability and IC(50) of NaAsO(2) on Lx-2 was then calculated; According to IC(50) results, 0.000, 1.875, 3.750, 7.500, and 15.000 µmol/L of NaAsO(2) were exposed to Lx-2 cells for 24 hours, besides, 7.500 µmol/L of NaAsO(2) was exposed to Lx-2 cells for 0, 12, 24, 48, and 72 hours, then collected cells and culture supernatant; HSC activation-related protein, including α-smooth muscle actin (α-SMA), and transforming growth factor-ß1 (TGF-ß1) expression levels were detected by Western blot analysis, the main extracellular matrix including laminin (LN) , hyaluronic acid (HA), collagen â £ (COL-â £) and procollagen â ¢(P â ¢ NP) secretion level was detected by Elisa assay. Results: CCK-8 assay showed that the cell viability of Lx-2 cells were increased obviously at low doses (≤1.0 µmol/L) of arsenic exposure, especially at 48 and 72 h. In contrast, with the increasing doses of arsenic exposure, the survival rate of Lx-2 cell was decreased gradually, and the survival rate of the high-dose (50, 100 µmol/L) arsenic exposure group at 24, 48 and 72 h were significantly lower than 0.0 µmol/L group, P<0.05. The IC(50) of NaAsO(2) on Lx-2 cells at 24, 48, 72 h were calculated as 72.75, 48.19 and 29.95 µmol/L, respectively; The expression levels of HSC activation-related protein showed that, after treated with 1.875, 3.750, 7.500, 15.000 µmol/L NaAsO(2) for 24 h, α-SMA and TGF-ß1 protein level were higher than 0.000 µmol/L group. The increased expression of α-SMA and TGF-ß1 protein were most significant in 7.500 µmol/L NaAsO(2) group (P<0.05). In addition, the expression levels of α-SMA and TGF-ß1 also showed a time-dependent increasing in Lx-2 cells after treated with 7.500 µmol/L NaAsO(2) for 0, 12, 24, 48 and 72 h; Elisa assay showed that after treated with 1.875, 3.750, 7.500, 15.000 µmol/L NaAsO(2) for 24 h, the secretion levels of HA, LN, COL-â £ and Pâ ¢NP were obvious higher than 0.000 µmol/L group (P<0.05). Moreover, the secretion levels of HA, LN, COL-â £ and P â ¢ NP also showed a time-dependent increased manner in Lx-2 cells after exposed to 7.500 µmol/L NaAsO(2) for 0, 12, 24, 48 and 72 h (P<0.05). Conclusion: NaAsO(2) exposure to Lx-2 cells can upregulate the expression level of HSC activation-related proteins, induce its further activation, then increase ECM secretion level.
Subject(s)
Arsenites/metabolism , Extracellular Matrix/metabolism , Hepatic Stellate Cells/metabolism , Sodium Compounds/metabolism , Arsenites/toxicity , Environmental Exposure/adverse effects , Humans , Liver Cirrhosis/chemically induced , Sodium Compounds/toxicityABSTRACT
BACKGROUND: Sudden cardiac death (SCD) occurs in a broad spectrum of cardiac pathologies and is an important cause of mortality in the general population. Idiopathic ventricular fibrillation (IVF) is a rare but important factor resulting in SCD. It is diagnosed in a resuscitated cardiac arrest victim underlying unknown cause, with documented ventricular fibrillation. Previous studies have demonstrated that mutations in dipeptidyl aminopeptidase-like protein-6 (DPP6) and cardiac sodium channel Nav1.5 (SCN5A) are the most important genetic factors involve in IVF. AIM: By using whole sequencing to identify the genetic lesion of a family with suspicious idiopathic ventricular fibrillation. DESIGN: Prospective genetic study. METHODS: In this study, we employed whole-exome sequencing in combination with arrhythmia-related gene filtering to identify the genetic lesion for a family suffering from suspicious IVF, syncope and SCD. We then generated the plasmids of DPP6-pcDNA3.1+ (WT and c.1578G>C/p.Q526H). Kv4.3-pcDNA3.1+ was co-transfected together with/without DPP6-pcDNA3.1+ (WT and/or c.1578G>C/p.Q526H) into HEK293 cells to perform the patch clamp experiments. RESULTS: A novel missense mutation (c.1578G>C/p.Q526H) of DPP6 was identified and co-segregated with affected patients in this family. Patch clamp experiments suggested that this novel mutation might result in a gain of function and disturb the efflux of potassium ion. CONCLUSION: Our study not only reported the second missense mutation of DPP6 in heart disease and expanded the spectrum of DPP6 mutations, but also contribute to the genetic diagnosis and counseling of families with suspicious IVF, syncope and SCD.
Subject(s)
Dipeptidyl-Peptidases and Tripeptidyl-Peptidases/genetics , Mutation, Missense , Nerve Tissue Proteins/genetics , Potassium Channels/genetics , Ventricular Fibrillation/diagnosis , Ventricular Fibrillation/genetics , Death, Sudden, Cardiac/etiology , Electrocardiography, Ambulatory , Family , Humans , Male , Middle Aged , Prospective Studies , Exome SequencingABSTRACT
Objective: To investigate the relationships between the expression of programmed death 1 (PD-1) and the epidermal growth factor receptor (EGFR) gene mutations in non-small cell lung cancer (NSCLC). The study also attempted to investigate the clinicopathological features and prognosis in NSCLC patients. Methods: The expression of PD-1 protein in 88 cases of NSCLC tumor tissues and adjacent tissues was detected by immunohistochemistry. The mutations of EGFR in NSCLC were detected by Polymerase Chain Reaction-Amplification Refractory Mutation System(PCR-ARMS) method. The expression of PD-1 and patients' clinical characteristics and prognosis were analyzed. Results: PD-1 was positive in 63.6%(56/88) NSCLC tumor tissues, which was significantly higher than that in adjacent normal tissues (21.6%, 19/88) (P<0.05). EGFR gene mutations were found in 43 cases (48.9%), in which 30 cases (69.8%)were PD-1 positive expression. 45 cases had the wild types of EGFR gene, in which 26 cases (57.8%) were PD-1 positive. There were 24 cases of 19Del EGFR mutations, including 20 cases (83.3%) of PD-1 positive expression. 19 patients had 21L858 EGFR mutations, including 10 cases (52.6%) of PD-1 positive expression. The expression of PD-1 in NSCLC was related to patients' smoking status, lymph node metastasis and EGFR gene mutations (P<0.05). The median progression-free survival time of patients with PD-1 positive and negative expression was 7.03 and 18.66 months, respectively (P=0.007). In patients with wild-type EGFR gene, the median progression-free survival time of PD-1 positive and negative expression was 25.21 and 38.24 months, respectively. The difference was statistically significant (P=0.024). The median progression-free survival time in 43 cases of EGFR mutant patients with PD-1 positive and negative expression was 21.23 and 31.44 months. The difference was not statistically significant (P=0.128). Conclusions: PD-1 expresses in both EGFR mutant and wild-type NSCLC, and its expression levelis different with various EGFR mutations. The expression of PD-1 in NSCLC is related to the prognosis of patients, and the prognosis of patients with positive PD-1 expression was poor.
Subject(s)
Carcinoma, Non-Small-Cell Lung/metabolism , Genes, erbB-1/genetics , Lung Neoplasms/metabolism , Mutation , Programmed Cell Death 1 Receptor/metabolism , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/mortality , Disease-Free Survival , Female , Humans , Lung Neoplasms/genetics , Lung Neoplasms/mortality , Lymphatic Metastasis , Male , Polymerase Chain Reaction , Prognosis , Time FactorsABSTRACT
This study explored the expression of interleukin 17D (IL-17D) secreted by human ovariancarcinoma cells and the effect of exogenous IL-17D transfection on MICA, which is the ligand of NKG2D, on the surface of ovary carcinoma cells. Human ovarian papillary serous adenocarcinoma cell line SKOV3, empty vector control cell line SKOV3/vector, exogenous human IL-17D stable-transfected cell line SKOV3/IL-17D, as well as cisplatin (CDDP)-resistant cell SKOV/CDDP were cultured; ovarian adenocarcinoma cell line OVCAR-3, empty vector control cell line OVCAR3/vector and OVCAE3/IL- 17D were observed under a microscope. In the study, methyl-thiazolyl-tetrazolium (MTT) method was used to detect the inhibition rate, resistance index and proliferation of SKOV3 and SKOV3/CDDP. It was found that the expression of IL-17 D in SKOV3/CDDP was much higher than that of its parent cell line SKOV3; IL-17D might be correlated to the drug resistance of cells; the proliferation of SKOV3 transfected with IL-17D was significantly accelerated, indicating IL-17D may be effective in promoting the growth of oncocyte.
Subject(s)
Adenocarcinoma/pathology , Immunity, Innate/drug effects , Interleukin-17/physiology , Ovarian Neoplasms/pathology , Adenocarcinoma, Papillary/pathology , Cell Division , Cell Line, Tumor , Cisplatin/pharmacology , Cystadenocarcinoma, Serous/pathology , Drug Resistance, Neoplasm , Drug Screening Assays, Antitumor , Female , Gene Expression Regulation, Neoplastic , Histocompatibility Antigens Class I/biosynthesis , Histocompatibility Antigens Class I/genetics , Humans , Interleukin-17/genetics , Neoplasm Proteins/biosynthesis , Neoplasm Proteins/genetics , Recombinant Fusion Proteins/metabolism , TransfectionABSTRACT
INTRODUCTION: Detection of leukemogenic fusion transcripts in acute myeloid leukemia (AML) is critical for AML diagnosis. NanoString nCounter system is a novel probe-based gene expression platform capable of measuring up to 800 targets with advantages of reproducibility, accuracy, and sample type flexibility. To study the potential application of NanoString in leukemia at clinic, we used this technology to detect AML leukemogenic fusion transcripts and compared the performances with clinical molecular assays. METHODS: We developed a NanoString assay to detect seven leukemogenic fusion transcripts, namely RUNX1-RUNX1T1 (e5e12), PML-RARA (bcr1, bcr2, and bcr3), and CBFB-MYH11 (e5e12, e5e8, and e5e7). We set up the cut-off value for each fusion transcript and tested 42 de novo AML samples. We compared the results with reverse transcriptase-polymerase chain reaction (RT-PCR) and TaqMan reverse quantitative-polymerase chain reaction (RQ-PCR), the molecular methods standardly used at clinic. RESULTS: We demonstrated that the NanoString and RT-PCR results correlate well (P < 0.0001) and are highly concordant (95.2%). Using TaqMan RQ-PCR as a validation method and gold standard, we demonstrated superior accuracy and sensitivity of NanoString compared to RT-PCR and comparable specificity. Furthermore, we showed that NanoString is not as sensitive as TaqMan RQ-PCR in detecting very low level of fusion transcripts. CONCLUSIONS: NanoString can serve as a reliable and alternative molecular method to multiplexed RT-PCR for diagnosis of de novo AML with the perspective of screening/quantitation of a large number of leukemogenic fusion transcripts and prognostic genes. However, NanoString may not be an alternative method for monitoring minimal residual disease in AML.