【摘要】 目的:探讨长链非编码RNA-NEAT1(lncRNA-NEAT1)通过与miR-34a相互作用影响肾癌细胞生长的机制。方法:检测60例肾癌患者癌组织、癌旁组织中miR-34a及NEAT1表达水平的差异。稳定培养A498细胞,转染si-NEAT1,并用脂质体法瞬时转染miR-34a inhibitor,敲低miR-34a或NEAT1的表达,检测对A498细胞生长活性的影响。将20只SD大鼠随机分为NC组、miR-34a inhibitor组,检测大鼠各项指标差异。结果:与癌旁组织相比,肾癌组织中NEAT1的表达水平显著降低(P<0.05),miR-34a表达水平明显升高(P<0.05)。转染si-NEAT1后,A498细胞的增殖能力、迁移能力和侵袭能力显著升高,细胞凋亡率明显降低。Western blotting检测显示,si-NEAT1组细胞的Ki-67蛋白、Vimentin蛋白相对表达量明显降低(P<0.05),Cleaved caspase-3蛋白、N-cadherin蛋白相对表达量明显升高(P<0.05)。与NC组相比,miR-34a inhibitor组大鼠肿瘤生长受到明显抑制,直径明显较小(P<0.05)。HE染色检测显示,miR-34a inhibitor组大鼠肺内转移病灶更少,直径更小。共转染miR-34a inhibitor和si-NEAT1后,si-miR-34a inhibitor的抗肿瘤效应被部分抵消,细胞迁移能力及侵袭能力显著提高。与单转染组比较,共转染组细胞的Ki-67蛋白、Vimentin蛋白和N-cadherin蛋白相对表达量明显升高(P<0.05),Cleaved caspase-3蛋白相对表达量明显降低(P<0.05)。结论:lncRNA-NEAT1可通过靶向作用于miR-34a进而调节Vimentin表达,从而抑制肾癌生长,有望成为一个新的治疗靶点。更多还原

【Abstract】 Objective:To investigate the mechanism of the interaction between lncRNA-NEAT1 and miR-34 a on the growth of RCC.Methods:The expression of miR-34 a and NEAT1 in 60 cases of renal carcinoma and adjacent tissues were detected.A498 cells were cultrued stably and trasfect with si-NEAT1.Transient transfection with miR-34 a inhibitor was performed by liposome method and the expression of miR-34 a or NEAT1 was knocked down to detect the effect on the growth activity of A498 cells.Twenty SD rats were randomly divided into NC group and miR-34 a inhibitor group and the differences of various indexes were detected.Results:The expression of lncRNA-NEAT1 was significantly decreased(P<0.05),and the expression of miR-34 a was significantly increased(P<0.05).After si-NEAT1 transfection, the proliferation, migration and invasion of A498 cells increased significantly, and the apoptosis rate decreased significantly.In si-NEAT1 group, the relative expression of Ki-67 protein and Vimentin protein decreased(P<0.05),while the expression of Cleaved caspase-3 protein and N-cadherin protein increased(P<0.05).Compared with NC group, the tumor growth of miR-34 a inhibitor group was significantly inhibited with a significantly smaller diameter(P<0.05).HE staining showed that, the number of pulmonary metastasis was less and the diameter was smaller.After go-transfection of miR-34 a inhibitor and si-NEAT1,the antitumor effect of si-miR-34 a was partially offset, and the cell migration and invasion ability were significantly improved.Compared with the single transfection group, the relative expression of Ki-67 protein, Vimentin protein and N-cadherin protein in go-transfection group was significantly increased(P<0.05),and the relative expression of Cleaved caspase-3 protein was significantly decreased(P<0.05).Conclusion:lncRNA-NEAT1 can target miR-34 a and regulate Vimentin expression, thus inhibiting the growth of renal cell carcinoma, which is expected to become a new therapeutic target.更多还原