本文關(guān)鍵詞：慢病毒介導(dǎo)靶向P27RF-Rho基因沉默抑制肝癌細(xì)胞侵襲性的實(shí)驗(yàn)研究　出處：《吉林大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： P27RF-Rho 慢病毒干擾載體 肝癌細(xì)胞 侵襲轉(zhuǎn)移

【摘要】：目的:應(yīng)用慢病毒介導(dǎo)特異性沉默P27RF-Rho,研究該基因?qū)Ω伟┘?xì)胞侵襲、遷移方面的影響。方法:構(gòu)建P27RF-Rho RNAi慢病毒載體。慢病毒介導(dǎo)感染肝癌細(xì)胞BEL7402。實(shí)驗(yàn)分3組:BEL7402空白對(duì)照組,Scramble-si RNA陰性對(duì)照組,P27RF-Rho-si RNA實(shí)驗(yàn)組。Western blot檢測(cè)P27RF-Rho基因沉默效果及Rho A、Rho C、VEGF、P53、PTEN表達(dá)水平;劃痕實(shí)驗(yàn)檢測(cè)細(xì)胞遷移能力、Transwell小室檢測(cè)細(xì)胞侵襲能力;明膠酶譜實(shí)驗(yàn)分析基質(zhì)金屬蛋白酶活性。結(jié)果:Western Blot顯示P27RF-Rho-si RNA實(shí)驗(yàn)組中P27RF-Rho蛋白表達(dá)水平低于兩個(gè)對(duì)照組(P0.05),BEL7402空白對(duì)照組,Scramble-si RNA陰性對(duì)照組中VEGF、Rho A、Rho C在蛋白水平上表達(dá)均高于實(shí)驗(yàn)組(P0.05),P27RF-Rho-si RNA實(shí)驗(yàn)組中P53、PTEN表達(dá)水平高于兩個(gè)對(duì)照組(P0.05);明膠酶譜實(shí)驗(yàn)發(fā)現(xiàn)P27RF-Rho-si RNA實(shí)驗(yàn)組細(xì)胞中的基質(zhì)金屬蛋白酶活性低于兩個(gè)對(duì)照組(P0.01);細(xì)胞劃痕實(shí)驗(yàn)顯示P27RF-Rho-si RNA實(shí)驗(yàn)組細(xì)胞遷移能力受到抑制(P0.01);P27RF-Rho-si RNA實(shí)驗(yàn)組穿過Transwell小室生物膜的細(xì)胞平均數(shù)低于兩個(gè)對(duì)照組,說明肝癌細(xì)胞的侵襲能力下降(P0.01)。結(jié)論:慢病毒介導(dǎo)特異性沉默P27RF-Rho,抑制肝癌細(xì)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(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.7

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