MicroRNA-188在胃癌中的表达及对胃癌细胞增殖和凋亡的影响

Expression of MicroRNA-188 in gastric carcinoma and its effect on the proliferation and apoptosis of gastric cancer cells

目的:探究microRNA-188在胃癌患者的组织和血液中的表达情况,并研究microRNA-188对人胃癌细胞MGC-803增殖和凋亡的影响及其机制。方法:收集10例胃癌患者的肿瘤组织和癌旁组织,应用PCR方法检测microRNA-545、microRNA-575、microRNA-1269、microRNA-99a、microRNA-452、microRNA-188的表达水平;收集健康志愿者和胃癌患者的血液,应用PCR检测这6种microRNAs的表达水平;应用转染试剂将microRNA-188 mimics、microRNA-188 AMO及其NC转染人胃癌细胞MGC-803, 24h后应用CCK-8实验和TUNEL染色检测细胞增殖能力和凋亡情况;应用在线网站,预测microRNA-188的靶基因;应用q-PCR和Western blot实验检测microRNA-188对SIX1是否具有调控作用。结果:与癌旁组织相比, microRNA-188在胃癌组织中表达显著降低。与健康人相比,microRNA-188在胃癌患者的血液中显著降低。CCK-8结果显示,microRNA-188 mimics显著抑制胃癌细胞的增殖,而microRNA-188 AMO能够显著提高胃癌细胞的增殖能力。TUNEL染色结果显示,microRNA-188能够促进胃癌细胞发生凋亡,而其AMO能够显著抑制胃癌细胞发生凋亡。q-PCR和Western blot实验结果表明,microRNA-188能够显著降低其靶点SIX1的mRNA和蛋白水平。结论:MicroRNA-188在胃癌患者的组织和血液中的表达水平显著降低,且其通过抑制SIX1发挥调控胃癌细胞增殖和凋亡的作用。

Objective: To investigate the expression of microRNA-188 in tissues and blood of gastric cancer patients, and to study the effect of microRNA-188 on the proliferation and apoptosis of human gastric cancer cell line MGC-803 and its mechanism. Methods: Gastric cancer tissues and adjacent tissues were collected from 10 patients with gastric cancer. Quantitative real time PCR(qPCR) was used to detect the expression levels of microRNA-545, microRNA-575, microRNA-1269, microRNA-99a, microRNA-452 and microRNA-188. The levels of six microRNAs in blood samples were detected by qPCR in healthy volunteers and patients with gastric cancer. The microRNA-188 mimics, microRNA-188 AMO and their NCs were transfected into human gastric cancer cells MGC-803 using transfection reagent. After 24 h, CCK-8 assay and TUNEL staining were used to detect the cell proliferation and apoptosis. The website online was used to predict the target gene of microRNA-188. qRT-PCR and western blot assays were used to determine whether microRNA-188 had a regulatory effect on SIX1. Results: Compared with adjacent tissues, microRNA-188 was found to markedly decrease in gastric cancer tissues. Compared with healthy volunteers, microRNA-188 was significantly lower in the blood samples of gastric cancer patients. CCK-8 results showed that microRNA-188 mimics significantly inhibited the proliferation of gastric cancer cells, while microRNA-188 AMO could significantly increase the proliferation of gastric cancer cells. TUNEL staining showed that microRNA-188 can promote the apoptosis of gastric cancer cells, and its AMO can significantly inhibit the apoptosis of gastric cancer cells. qRT-PCR and western blot experiments showed that microRNA-188 can significantly reduce the mRNA and protein levels of its target SIX1. Conclusion: The expression level of microRNA-188 is significantly decreased in the tissue and blood of patients with gastric cancer and microRNA-188 plays an important role in regulating the proliferation and apoptosis of gastric cancer cells by inhibiting SIX1.