论文部分内容阅读
目的研究胃癌和癌旁组织中microRNA-582(miR-582)的表达水平,以及调控miR-582表达后对胃癌细胞SGC-7901功能的影响。方法采用实时荧光定量PCR检测人胃癌和正常胃组织中miR-582的表达水平;将miR-582 inhibitors(miR-582下调表达)通过脂质体转染至人胃癌SGC-7901细胞中(miR-582 inhibitors组),设未转染对照组(control组)和miRNA无义序列转染对照组(NC组)。实时荧光定量PCR检测转染组细胞中miR-582的表达变化;MTT方法检测各组细胞的生长活力;流式细胞术检测各组的细胞周期和凋亡率;Transwell实验检测各组细胞侵袭能力;Western blot检测各组细胞中Rab7a、CDK1和AKT3蛋白表达。结果 miR-582在胃癌中的表达量为癌旁组织的(0.34±0.11)倍(P<0.05)。与control组和NC组相比,miR-582 inhibitors组中SGC-7901细胞增殖和侵袭能力上升(P<0.05),细胞周期被促进,细胞凋亡减少(P<0.05),Rab23蛋白表达上调,差异具有统计学意义(P<0.05)。结论 miR-582在胃癌组织中低表达,miR-582通过下调Rab23抑制胃癌细胞SGC-7901增殖、侵袭、迁移能力,促进其凋亡。其可以作为胃癌诊断和治疗的新靶点。
Objective To investigate the expression of microRNA-582 (miR-582) in gastric cancer and adjacent non-cancerous tissues and the effect of miR-582 on the function of gastric cancer SGC-7901. Methods Real-time quantitative PCR was used to detect the expression of miR-582 in human gastric cancer and normal gastric tissues. MiR-582 inhibitors (miR-582 down-regulated) were transfected into human gastric cancer cell line SGC-7901 by miR- 582 inhibitors group), untransfected control group and miRNA nonsense sequence control group (NC group). Real-time quantitative PCR was used to detect the expression of miR-582 in transfected cells. MTT assay was used to detect cell viability. Flow cytometry was used to detect the cell cycle and apoptosis rate. Transwell assay was used to detect the invasion ability The protein expressions of Rab7a, CDK1 and AKT3 in each group were detected by Western blot. Results The expression level of miR-582 in gastric cancer was (0.34 ± 0.11) times (P <0.05). Compared with control group and NC group, the proliferation and invasion ability of SGC-7901 cells in miR-582 inhibitor group were increased (P <0.05), the cell cycle was promoted, the apoptosis was decreased (P <0.05), Rab23 protein expression was up- The difference was statistically significant (P <0.05). Conclusion miR-582 is down-regulated in gastric cancer tissues. MiR-582 can inhibit the proliferation, invasion and migration of gastric cancer SGC-7901 cells by down-regulating Rab23. It can be used as a new target for the diagnosis and treatment of gastric cancer.