MicroRNA-7-5p 调控p53 表达对非小细胞肺癌 增殖及侵袭能力的影响

Effect of miR-7-5p on the proliferation, invasion and metastasis of non-small cell lung cancer

摘要:

目的 探讨microRNA-7-5p（miR-7-5p）是否通过调控p53 基因的表达而影响人非小细胞肺 癌（NSCLC）细胞体外的增殖、侵袭能力。方法 通过双荧光素酶报告证明p53 为miR-7-5p 的下游靶基 因，将miR-7-5p 转染至人NSCLC 细胞株NCI-H1975，依据实验对照原则分为空白对照组、miR-7-5p 组 和miR-NC 组，观察细胞中p53 基因在转录和蛋白水平的表达变化；然后通过细胞增殖、细胞周期实验及 侵袭实验，观察NCI-H1975 细胞体外增殖和侵袭能力的变化。结果 miR-7-5p 组的p53 mRNA 相对表达 量、迁移细胞数及克隆形成数均低于miR-NC 组（P <0.05）。miR-7-5p 组3'UTR 荧光素酶的表达活性较 miR-NC 组低（P <0.05）。miR-7-5p 组p53 mRNA 和蛋白相对表达量较miR-NC 组低（P <0.05）。miR- 7-5p 组细胞增殖能力较miR-NC 组低（P <0.05）。miR-7-5p 组细胞周期G0/G1 期比例高于空白对照组 和miR-NC 组（P <0.05）。miR-7-5p 组细胞周期的G2/M 期比例低于miR-NC 组（P <0.05）。miR-7- 5p 组细胞周期的S 期比例低于空白对照组和miR-NC 组（P <0.05）。miR-7-5p 组细胞侵袭能力低于空白对 照组和miR-NC 组（P <0.05）。结论 miR-7-5p 主要是通过靶向p53 促进其mRNA 和蛋白的表达，并减弱人 NSCLC 的增殖和侵袭能力。

Abstract:

Objective To investigate whether the expression of p53 regulated by miR-7-5p affects the proliferation and invasion of human non-small cell lung cancer. Methods p53 was validated as a downstream target gene of miR-7-5p by dual luciferase reporter assay. Human non-small cell lung cancer cell line NCI-H1975 was transfected with miR-7-5p, and control group, miR-7-5p group and miR-NC group were established to observe level changes of p53 mRNA and protein detected by RT-RCR and Western blot. In addition, our group observed the proliferation and invasion of NCI-H1975 cells through MTS cell proliferation assay, cell cycle assay and Transwell invasion assay. Results In the lung cancer cell line (NCI-H1975), the expression of p53 mRNA, the number of migrated cells and the number of clone formation in the miR-7-5p group were lower than those in the miR-NC group (P < 0.05). The miR-7-5p group reduced luciferase expression activity compared to the miR-NC group (P < 0.05).Compared with the miR-NC group, the expression of p53 protein in the miR-7-5p group was lower (P < 0.05). The cell proliferation ability of miR-7-5p group was lower than that of miR-NC group (P < 0.05). The G0/G1 (%) ratio of the cells in the miR-7-5p group was higher than that of the control group and the miR-NC group (P < 0.05); the G2/M (%) ratio of cells in the miR-7-5p group was lower than that in the miR-7-5p group (P < 0.05); the S (%) ratio of the cells in the miR-7-5p group was lower than that in the control group and the miR-NC group (P < 0.05). The invasive ability of miR-7-5p group was lower than that of the control group and miR-NC group (P < 0.05). Conclusions The miR-7-5p promotes the expression of p53 mRNA and protein to suppress of the proliferation and invasion of human non-small cell lung cancer.