mir-145对hepg2细胞侵袭迁移的影响及可能机制word论文.docxVIP

miR-145 对 HepG2 细胞侵袭迁移的影响及可能机制硕士研究生：李健辉 导师：陈宏辉 中文摘要目的探讨 miR-145 对肝癌细胞 HepG2 侵袭迁移的影响及可能机制，为 miRNA 最终 成为肝癌治疗的新靶点奠定基础。方法体外培养人肝癌细胞 HepG2；脂质体介导的转染方法将 miR-145-5p 模拟物（miR-145-5p mimics）转染 HepG2 细胞，同时设立 Blank 组（空白对照组）、Mock 组（转染试剂对照组）、NC 阴性对照组（无关干扰序列阴性对照组）；利用生物信 息学软件预测 miR-145 潜在的靶基因；采用 cck-8 法检测细胞的增殖能力；通过 Transwell 侵袭实验和划痕实验检测细胞的侵袭和迁移能力；QRT-PCR 检测转染 miR-145-5p mimics 后细胞中 miR-145、MUC1 mRNA 的表达；Western-blot 检测转 染 miR-145-5p mimics 后 MUC1 蛋白的表达。结果1.CCK-8 结果显示 miR-145-5p mimics 组的细胞增殖活性明显低于空白对照组、 转染试剂对照组及无关干扰序列阴性对照组。（P﹤0.05）。Transwell 侵袭实验显示转染 miR-145-5p mimics 72 h（即细胞接种 48h）后， miR-145-5p mimics 组侵出小室的细胞数较空白对照组、无关干扰序列阴性对照组和 转染试剂组明显减少，且有统计学意义（P﹤0.05）。3.划痕实验显示划痕后 24 h 及 48h ，miR-145-5p mimics 组 HepG2 细胞迁移能力 较空白对照组、转染试剂组和无关干扰序列阴性对照组明显下降（P﹤0.05）。RT-PCR 结果表明转染 miR-145-5p mimics 48 小时后，miR-145-5p 的 mRNA表达水平明显上升（P﹤0.05）,MUC1mRNA 表达水平无明显变化（P﹥0.05）。2Western-blot 结果表明转染 miR-145-5p mimics 48 小时后,MUC1 的蛋白表达水平下降（P﹤0.05）。结论1.miR-145 对肝癌细胞 HepG2 的增殖和侵袭迁移存在负性调控作用。2.miR-145 可能通过作用于靶基因 MUC1 抑制肝癌细胞 HepG2 的侵袭迁移。关键词肝癌；miR-145；侵袭；迁移 ； MUC13Effect and possible mechanism of miR-145 on the invasion andmigration of HepG2 cellsAbstractObjectiveThis study is to investgate the effect of miR-145 on the invasion and migration of HepG2 cells and the possible mechanisim，in order to lay a foundation for the treatment of liver cancer.MethodsHepG2 cells were cultured in vitro and transfected with miR-145 mimics.At the same time we set up blank control group,transfection reagent group,and NC negative control group.Bioinformatics software forecasted the potential target genes of miR-145.The proliferation of the HepG2 was evaluated by CCK-8 assay. Invasion and cell migrating ability were evaluated by transwell invasion assay and scratch assay.After HepG2 cells were transfected with miR-145 mimics,the mRNAs of miR-145 and MUC1 were detected through QRT-PCR and the protein expression of MUC1 was detected through western- blot analysis.ResultsCCK-8 method showed that cells transfected with miR-145 mimics revealed si