| Objective:The aim was to research the expression of microRNA-106b-25 gene cluster and relationship with clinicopathological features in gastric cancer. At the same time, we detected the expression levels of miRNA-106b-25 in gastric cancer cell lines using transient transfection technique to suppress the expression of miRNA-106b-25 in vitro, and explored the effects of behavioral characteristics in low-miRNA-106b-25 expression levels in gastric cancer cell lines. Methods:1. QRT-PCR was used to detect relative expression levels of mi RNA-106b-25 in 90 tissues of gastric cancer and adjacent normal gastric mucosa. We collected 90 patients with gastric cancer who were hospitalized into Tianjin Medical University Cancer Hospital from March 2013 to August 2013, and analyzed the difference between gastric cancer tissues and their adjacent normal gastric mucosa. The clinical and pathological features of all enrolled patients were also checked, and furtherly analyzed the correlation between miRNA-106b-25 and these clinicopathological features.2. The same method was used to detect expression of miRNA-106b-25 in gastric cancer cell line SGC-7901, MGC-803, BGC-823 and GES-1. Meanwhile, synthetic miRNA-106b-25 inhibitor were transiently transfected in cell lines with lipo2000, and we observed transfection efficiency by fluorescence microscopy and qRT-PCR; then selected gastric cancer cell lines with best inhibitory effect as subjects for further study.3. After transfection, we used scratch test to observe migration of gastric cancer cell, growth curve and MTT method to observe changes of proliferation and growth inhibition rate, PI and Annexin V / PI staining to explore changes of cell cycle and apoptosis and Transwell test for invasion ability. Results:1. The miRNA-106b-25 expression in gastric cancer tissues was significantly higher than in adjacent normal gastric mucosa. MiRNA-106 b and miRNA-93 relative expression were significantly associated with tumor size, serous invasion, vascular invasion, depth of invasion, lymph node metastasis, distant metastasis and TNM stage; miRNA-25 was only were associated with serous invasion, vascular invasion, depth of invasion and lymph node metastasis.2. The expression of miRNA-106 b, miRNA-93 and mi RNA-25 in gastric cancer cell line SGC-7901, MGC-803, BGC-823 was higher than normal gastric cell line GES-1. Compared to the other two kinds of gastric cancer cell line(SGC-7901, BGC-823), MGC-803 owned a better inhibitory effect with miRNA inhibitor, so we chose MGC-803 cell as subject for function research.3. After transiently transfected miRNA inhibitor into MGC-803, the scratch test and Transwell experiments show that cell migration and invasion decreased significantly; MTT test and cell growth curve was proliferation significantly inhibited; Annexin V / PI method validated increased apoptosis in MGC-803. Conclusion:1. MiRNA-106b-25 gene cluster overexpressed in gastric carcinoma, and the expression of this cluster miRNA was significant correlated with clinicopathological features which closely related were prognosis. It suggested that its expression could be used as an important clinical indicators to reflect the progression of gastric cancer.2. Relative to normal gastric mucosa cells, mi RNA-106b-25 gene clusters were overexpressed in several gastric cancer cell lines; and after suppression by exogenous inhibitors, a number of biological behavior were significantly changed. It suggested that miRNA-106b-25 might influence the key parts in the development process with gastric cancer, and achieved a solid foundation for further exploration to find key molecular targets for treatment. |
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