慢病毒介导靶向P27RF-Rho基因沉默抑制肝癌细胞侵袭性的实验研究

发布时间：2018-01-03 05:27

  本文关键词：慢病毒介导靶向P27RF-Rho基因沉默抑制肝癌细胞侵袭性的实验研究　出处：《吉林大学》2017年硕士论文　论文类型：学位论文

  更多相关文章： P27RF-Rho 慢病毒干扰载体 肝癌细胞 侵袭转移

【摘要】：目的:应用慢病毒介导特异性沉默P27RF-Rho,研究该基因对肝癌细胞侵袭、迁移方面的影响。方法:构建P27RF-Rho RNAi慢病毒载体。慢病毒介导感染肝癌细胞BEL7402。实验分3组:BEL7402空白对照组,Scramble-si RNA阴性对照组,P27RF-Rho-si RNA实验组。Western blot检测P27RF-Rho基因沉默效果及Rho A、Rho C、VEGF、P53、PTEN表达水平;划痕实验检测细胞迁移能力、Transwell小室检测细胞侵袭能力;明胶酶谱实验分析基质金属蛋白酶活性。结果:Western Blot显示P27RF-Rho-si RNA实验组中P27RF-Rho蛋白表达水平低于两个对照组(P0.05),BEL7402空白对照组,Scramble-si RNA阴性对照组中VEGF、Rho A、Rho C在蛋白水平上表达均高于实验组(P0.05),P27RF-Rho-si RNA实验组中P53、PTEN表达水平高于两个对照组(P0.05);明胶酶谱实验发现P27RF-Rho-si RNA实验组细胞中的基质金属蛋白酶活性低于两个对照组(P0.01);细胞划痕实验显示P27RF-Rho-si RNA实验组细胞迁移能力受到抑制(P0.01);P27RF-Rho-si RNA实验组穿过Transwell小室生物膜的细胞平均数低于两个对照组,说明肝癌细胞的侵袭能力下降(P0.01)。结论:慢病毒介导特异性沉默P27RF-Rho,抑制肝癌细胞BEL7402的侵袭迁移。
[Abstract]:Objective: to study the invasion of P27RF-Rhoby lentivirus mediated specific silencing of P27RF-Rho. Methods: P27RF-Rho RNAi lentivirus vector was constructed. The cell line BEL7402 was infected by lentivirus. The experiment was divided into three groups:. BEL7402 blank control group. Scramble-si RNA negative control group. P27RF-Rho-si RNA group. Western blot was used to detect the silencing effect of P27RF-Rho gene. The expression level of P53 and PTEN; The ability of cell migration was detected by scratch test and the ability of cell invasion was detected by Transwell chamber. The activity of matrix metalloproteinases was analyzed by gelatinase assay. Results: P27RF-Rho-si was detected by Blot. The expression of P27RF-Rho protein in RNA group was lower than that in two control groups. P0.05). VEGFU Rho A was detected in BEL7402 blank control group and Scrale-si RNA negative control group. The expression of Rho C at protein level was higher than that of p53 in P0.05P27RF-Rho-si RNA group. The expression level of PTEN was higher than that of control group (P0.05A). The activity of matrix metalloproteinases in P27RF-Rho-si RNA experimental group was lower than that in two control groups. The cell scratch test showed that the migration ability of P27RF-Rho-si RNA experimental group was inhibited (P0.01). The average number of P27RF-Rho-si RNA cells passing through the Transwell chamber biofilm was lower than that of the two control groups. Conclusion: lentivirus-mediated specific silencing of P27RF-Rhoand inhibits the invasion and migration of hepatoma cell line BEL7402.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7

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