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目的检测胃癌组织和癌旁组织中miR-497的表达差异,寻找其下游作用靶点。方法选取2010~2013年期间在中国医科大学附属第一医院胃肠外科手术切除的94例胃癌及其癌旁组织。用RT-PCR法定量检测miR-497在其中的表达;在线软件预测miR-497靶基因,用阴性对照及miR-497-mimics转染SGC-7901细胞系,RT-PCR及Western检测胃癌细胞miR-497过表达后靶点基因mRNA及蛋白水平的改变;MTT法检测miR-497-mimics组(miR-497过表达)细胞株与阴性对照组间对5-氟尿嘧啶(5-FU)化疗的敏感性。结果①miR-497表达量在胃癌组织中明显低于癌旁组织,差异有统计学意义(P<0.01)。②BCL2L2被预测为miR-497的靶基因,miR-497在胃癌SGC-7901细胞中过表达后BCL2L2蛋白表达在miR-497-mimics组细胞中明显弱于阴性对照组细胞,BCL2L2 mRNA在2组细胞中的表达差异无统计学意义(P>0.05)。miR-497-mimics组细胞对5-FU的IC50值明显低于阴性对照组细胞(P=0.004 6)。结论 miR-497在胃癌组织中低表达,BCL2L2为miR-497的靶基因,miR-497过表达可提高胃癌细胞对5-FU的敏感性,这为进一步研究miR-497与BCL2L2在胃癌中的作用及治疗提供了新的方向。
Objective To detect the expression differences of miR-497 in gastric cancer tissues and paracancerous tissues, and to find out the downstream targets. Methods Totally 94 cases of gastric cancer and its peritumoral tissues resected from January 2006 to January 2013 in the First Affiliated Hospital of China Medical University were selected. The expression of miR-497 was quantitatively detected by RT-PCR. The target gene was predicted by online software. The negative control and miR-497-mimics were transfected into SGC-7901 cell line. RT-PCR and Western blotting were used to detect miR- -497 overexpression. The MTT assay was used to detect the sensitivity of 5-fluorouracil (5-FU) chemotherapy in miR-497-mimics group (miR-497 overexpression) and negative control group Sex. Results ① The expression of miR-497 in gastric cancer tissues was significantly lower than that in paracancerous tissues, the difference was statistically significant (P <0.01). The expression of BCL2L2 protein in miR-497-mimics group was significantly lower than that in the negative control group when miR-497 was overexpressed in gastric cancer SGC-7901 cells. ② The expression of BCL2L2 mRNA in the two groups of cells In the expression of no significant difference (P> 0.05). The IC50 values of 5-FU in miR-497-mimics group were significantly lower than those in negative control group (P = 0.004 6). CONCLUSIONS: miR-497 is low expressed in gastric cancer tissues and BCL2L2 is the target gene of miR-497. Overexpression of miR-497 can enhance the sensitivity of gastric cancer cells to 5-FU. This is the first study to investigate the role of miR-497 and BCL2L2 in gastric cancer Role and treatment provides a new direction.