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目的:探讨miR-203a-3p在胃癌中的表达及其对胃癌细胞增殖的影响。方法:收集胃癌组织标本44例,采用Real-time PCR方法检测胃癌组织标本中miR-203a-3p的表达,并分析其表达水平与临床病理参数的关系;生物信息学预测miR-203a-3p的靶基因,并采用荧光素酶报告基因实验进行验证;免疫组化检测胃癌组织标本中PRMT5的表达情况,分析其表达水平与miR-203a-3p表达的相关性;利用脂质体介导的瞬时转染方法过表达miR-203a-3p或同时过表达miR-203a-3p和PRMT5,并通过CCK-8实验检测胃癌细胞的增殖情况。结果:胃癌组织中miR-203a-3p的表达水平与正常组织对照相比显著降低(P<0.01),并且miR-203a-3p的表达水平与肿瘤细胞的分化程度显著相关;荧光素酶报告基因实验证实miR-203a-3p可以直接结合在PRMT5 3’-UTR上,即PRMT5是miR-203a-3p的直接靶基因;在胃癌组织标本中,PRMT5表达水平显著高于正常组织对照(P<0.01),并且其表达水平与miR-203a-3p表达呈显著负相关(r=-0.4124,P<0.01);过表达miR-203a-3p后,胃癌BGC823细胞的增殖能力显著低于miR-NC对照组(P<0.01),并且,“挽救”实验表明在过表达miR-203a-3p的细胞中同时过表达PRMT5后会部分恢复miR-203a-3p对细胞增殖的抑制(P<0.01)。结论:miR-203a-3p可通过下调靶基因PRMT5的表达,进而抑制胃癌细胞增殖。因此,miR-203a-3p可作为胃癌疾病临床治疗的潜在靶点。
Objective: To investigate the expression of miR-203a-3p in gastric cancer and its effect on the proliferation of gastric cancer cells. Methods: Forty-four gastric cancer specimens were collected. Real-time PCR was used to detect the expression of miR-203a-3p in gastric cancer tissues. The relationship between miR-203a-3p expression and clinicopathological parameters was analyzed by bioinformatics. The expression of PRMT5 was detected by immunohistochemistry and the correlation between the expression level of miR-203a-3p and the expression of miR-203a-3p was analyzed by immunohistochemistry. The liposome-mediated transient The transfection method overexpression of miR-203a-3p or overexpression of miR-203a-3p and PRMT5, and CCK-8 test detected gastric cancer cell proliferation. Results: The expression of miR-203a-3p in gastric cancer tissues was significantly lower than that in normal tissues (P <0.01), and the expression level of miR-203a-3p was significantly correlated with the differentiation of tumor cells. The luciferase reporter gene The results showed that miR-203a-3p could directly bind to 3’-UTR of PRMT5, that is, PRMT5 was a direct target of miR-203a-3p. In gastric cancer specimens, PRMT5 expression was significantly higher than that in normal tissues (P <0.01 ), And the expression of miR-203a-3p was significantly negatively correlated (r = -0.4124, P <0.01). The proliferation of BGC823 cells was significantly lower than that of the miR-NC control (P <0.01). Moreover, “rescue” experiments showed that the simultaneous suppression of miR-203a-3p cell proliferation by PRKT5 overexpression in miR-203a-3p-overexpressing cells (P <0.01) . Conclusion: miR-203a-3p can inhibit the proliferation of gastric cancer cells by down-regulating the expression of target gene PRMT5. Therefore, miR-203a-3p can be used as a potential target for the clinical treatment of gastric cancer.