mir-34a抑制人胃癌细胞生长与侵袭的实验研究word论文.docxVIP

miR-34a 抑制人胃癌细胞生长与侵袭的实验研究硕士研究生 ：陈艳辉 导师 ：罗招阳中文摘要目的：探讨 miR-34a（Homo sapiens miR-34a）在人胃癌 MGC-803 细胞中的生物 学功能以及分子机制研究。方法：采用 MTT、Transwell 侵袭实验、细胞划痕实验、AnnexinV-FITC/PI 染色 分别检测过表达 miR-34a 对人胃癌 MGC-803 细胞的生长、侵袭和凋亡作用；采 用 Western blot 检测 miR-34a 的靶基因 Survivin，E2F3，Bcl-2，CD44 蛋白的表 达。结果：MTT 检测发现，过表达 miR-34a 可抑制人胃癌 MGC-803 细胞的生长增殖， 具有时间依赖性；细胞划痕和 Transwell 侵袭实验显示，在胃癌 MGC-803 细胞中 过表达 miR-34a 36 h 后伤口愈合率为 (34.7±5.5) %，与对照组 (69.7±5.1) % 比 能明显减缓 MGC-803 细胞划痕伤口的愈合，转染 miR-34a mimics 36 h 后穿过基 底膜的细胞数为 (67±14)，与对照组 (154±12) 比能明显减缓 MGC-803 细胞侵袭 能力，差异均有统计学意义(P0.01)；AnnexinV-FITC/PI 实验显示，转染 miR-34a mimics 能显著促进胃癌 MGC-803 细胞的凋亡作用；Western blot 结果显示，过 表达 miR-34a 可抑制 Survivin，Bcl-2，CD44 蛋白的表达。结论：1. miR-34a 可抑制胃癌 MGC-803 细胞生长与侵袭，诱导胃癌 MGC-803 细胞凋 亡。2. miR-34a 可下调胃癌 MGC-803 细胞中 Survivin、Bcl-2、CD44 蛋白的表达。 关键词：miR-34a；胃癌；生长与侵袭；凋亡2The study of the miR-34a inhibiting the gastric cancer cells growth and invasionAbstractObjective: To analyze the mechanism and the role of miR-34a in the gastric cancer cells.Methods: Cell growth was evaluated by MTT. Cell invasion potential was evaluated by scratch test and transwell assay. The apoptosis cells were tested by AnnexinV-FITC/PI staining. Finally, western blotting was used to analyze the effects of miR-34a on Survivin, E2F3, Bcl-2, CD44 protein expression.Results: The MTT assay demonstrated that after overexpressing miR-34a in gastric cancer cells, cellular viability was significantly reduced (p0.05). The Transwell invasion chamber assay illustrated that after increasing the expression of miR-34a, the number of cells passing through the Transwell chamber were significantly reduced (p0.01). Flow cytometry analysis determined that upon miR-34a overexpression, the apoptosis cells increased significantly. Based on western blotting, compared with the control group, Survivin, Bcl-2, CD44 protein expression levels were significantly decreased in the gastric cancer cells transfected with the miR-34a mimic for 48 h. Conclusions:1.miR-34a can inhibit gastr