本文關鍵詞：慢病毒介導靶向P27RF-Rho基因沉默抑制肝癌細胞侵襲性的實驗研究　出處：《吉林大學》2017年碩士論文　論文類型：學位論文

  更多相關文章： P27RF-Rho 慢病毒干擾載體 肝癌細胞 侵襲轉移

【摘要】：目的:應用慢病毒介導特異性沉默P27RF-Rho,研究該基因對肝癌細胞侵襲、遷移方面的影響。方法:構建P27RF-Rho RNAi慢病毒載體。慢病毒介導感染肝癌細胞BEL7402。實驗分3組:BEL7402空白對照組,Scramble-si RNA陰性對照組,P27RF-Rho-si RNA實驗組。Western blot檢測P27RF-Rho基因沉默效果及Rho A、Rho C、VEGF、P53、PTEN表達水平;劃痕實驗檢測細胞遷移能力、Transwell小室檢測細胞侵襲能力;明膠酶譜實驗分析基質金屬蛋白酶活性。結果:Western Blot顯示P27RF-Rho-si RNA實驗組中P27RF-Rho蛋白表達水平低于兩個對照組(P0.05),BEL7402空白對照組,Scramble-si RNA陰性對照組中VEGF、Rho A、Rho C在蛋白水平上表達均高于實驗組(P0.05),P27RF-Rho-si RNA實驗組中P53、PTEN表達水平高于兩個對照組(P0.05);明膠酶譜實驗發(fā)現(xiàn)P27RF-Rho-si RNA實驗組細胞中的基質金屬蛋白酶活性低于兩個對照組(P0.01);細胞劃痕實驗顯示P27RF-Rho-si RNA實驗組細胞遷移能力受到抑制(P0.01);P27RF-Rho-si RNA實驗組穿過Transwell小室生物膜的細胞平均數(shù)低于兩個對照組,說明肝癌細胞的侵襲能力下降(P0.01)。結論:慢病毒介導特異性沉默P27RF-Rho,抑制肝癌細胞BEL7402的侵襲遷移。
[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.
【學位授予單位】：吉林大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R735.7

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