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Expression of HINT1 and Epigenetic Mechanism in Gastric Cancer

Author: HuangHuaYing
Tutor: ChenJinFei
School: Nanjing Medical University
Course: Oncology
Keywords: HINT1 Gastric cancer Gastric cancer cell lines PCR Western blot
CLC: R735.2
Type: Master's thesis
Year: 2011
Downloads: 54
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Abstract

Objective: To study the expression of HINT1 in gastric cancer, and the combination of clinical and pathological data analysis and its clinical significance; to explore possible epigenetic mechanisms the gastric HINT1 expression. Methods: real-time fluorescence quantitative polymerase chain reaction (Real-time Fluorescent Quantitative Poly-merase Chain Reaction, q-PCR) analysis of 34 patients with gastric cancer and its corresponding non-tumor tissue Hint1 expression levels on tumor group, non- The amount of gene expression in the tumor group statistical analysis; combined with clinical and pathological information, statistical analysis of relative gene expression correlation between the level of clinical and pathological features; while the RT-PCR method to detect gastric cancer and its corresponding non-tumor group Hp, and EB virus infection, and statistical analysis groups Hint1 relative gene expression levels between the infected group and non-infected group. The protein immunoblotting Western blot in 51 cases of gastric cancer and its corresponding protein level of non-tumor tissue HINT1; combination of clinical and pathological information, statistical analysis of relative gene expression levels between the clinical and pathological features; gastric cancer and its corresponding non-tumor group Hp EB virus infection grouping and statistical analysis HINT1 relative gene expression levels between the infected group and non-infected group. Immunohistochemical methods were used to detect the five cases of poorly differentiated gastric differentiation in five cases and the corresponding non-tumor tissue paraffin tissue sections HINT1 protein level, analysis of the correlation between the degree of differentiation and gene expression. Department of gastric cancer and normal gastric mucosa cells detected by RT-PCR and Western blot method HINT1 expression and epigenetic drug DNA methylation inhibitor 5 - azacytidine (5-Azacytidine 5-Aza - dC ) deal with low expression of gastric cancer cells, to detect the protein and mRNA expression level of recovery situations. Detection of gastric cancer and its corresponding non-tumor tissue Hint1 promoter region methylation status using methylation-specific PCR (Methylation Specific PCR, MSP). Results: 34 cases of gastric cancer, Hint1 expression in the tumor group was significantly lower than the non-tumor tissue (p = 0.03), 58.82% tumor the organization showed Hint1 low expression compared to the corresponding non-tumor tissue. Hp infection group, Hint1 expression in tumors was significantly lower than that of non-tumor tissue (p = 0.039); rather than infection group showed no significant difference; EB non-infected group, Hint1 in tumors was significantly lower than that of non-tumor tissue infected group showed no significant difference (p = 0.001); 51 cases of gastric cancer in the Western blot assay, 39.22% of the tumor the organization showed HINT1 low expression, compared to the corresponding non-tumor tissue and poorly differentiated group was significantly lower than in the well differentiated group (p = 0.027). The Hp, EB infected with at least one case, HINT1 low expression in tumor tissues trend more evident (p = 0.005). 10 patients with gastric cancer and corresponding non-tumor tissue paraffin sections immunohistochemical methods positioning the detection, HINT1 expression in the cytoplasm and nucleus, in the case of the corresponding non-tumor tissues showed positive expression, 4/5, poorly differentiated tumors performance low expression, 4/5 in the differentiation of tumor tissue showed positive expression. Consistent trend with the Western blot results. Low expression of gastric cancer cell lines AGS plus 5-Aza-dC for 72 h, and found that the expression of cell mRNA and protein levels HINT1 significantly restored, and there is a certain concentration-dependent. Typical cases found Western blot shown HINT1 the cases of inhibition is expressed in the tumor tissue, the results of the corresponding MSP shown the tumor tissue Hint1 promoter region for the methylation status of, and adjacent to the non-tumor tissue unmethylated state . Conclusion: In this study, the following conclusions: 1, at the mRNA level, Hint1 in gastric cancer showed low expression patterns; combined the Hp and EB infection, Hint1 gastric Hp infection group and EB non-infected group showed low expression patterns. 2, at the protein level, HINT1 performance in the gastric low expression pattern, its low expression in poorly differentiated group becomes more apparent, and by Hp infection affects EB. 3, the study found Hint1 low expression and DNA methylation may be related to low expression of gastric cancer cell lines. 4 gastric cancer MSP analysis is preliminary indication of the low Hint1 expression and methylation of the promoter region.

Expression of HINT1 and Epigenetic Mechanism in Gastric Cancer

Abstract

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