【摘要】：目的:探討慢病毒干擾載體沉默Yes-相關(guān)蛋白（Yes-associatedprotein, YAP）對(duì)人腎癌786-0細(xì)胞增殖和凋亡的影響。 方法:體外培養(yǎng)腎癌786-0細(xì)胞，構(gòu)建針對(duì)YAP基因的shRNA慢病毒干擾載體并轉(zhuǎn)染786-0細(xì)胞。RT-PCR和Western blot分別檢測(cè)干擾786-0細(xì)胞前后YAP及其下游轉(zhuǎn)錄因子TEAD1mRNA及蛋白的表達(dá)情況；CCK-8(cell counting kit-8)法檢測(cè)沉默YAP后細(xì)胞增殖的改變；流式細(xì)胞儀（flow cytometry, FCM）檢測(cè)細(xì)胞凋亡和周期的變化情況。 結(jié)果:構(gòu)建了3條針對(duì)YAP基因的YAP-shRNA慢病毒干擾載體和1條陰性對(duì)照慢病毒載體并成功轉(zhuǎn)染786-0細(xì)胞；YAP-shRNA慢病毒干擾載體轉(zhuǎn)染96小時(shí)后，可顯著下調(diào)786-0細(xì)胞YAP mRNA及蛋白表達(dá)水平(P=0.000)，并且TEAD1mRNA及蛋白表達(dá)水平也明顯下降(P=0.000)；與空白對(duì)照組和陰性對(duì)照組細(xì)胞相比，轉(zhuǎn)染組細(xì)胞增殖明顯抑制，細(xì)胞凋亡顯著增加(P=0.000)，細(xì)胞周期紊亂，，G1期細(xì)胞明顯增加，S期細(xì)胞明顯降低(P=0.000)。 結(jié)論: YAP-shRNA慢病毒干擾載體能有效抑制YAP基因在786-0細(xì)胞中表達(dá)，進(jìn)而抑制細(xì)胞增殖并促進(jìn)細(xì)胞凋亡。
[Abstract]:Aim: to investigate the effect of lentivirus interference vector silencing Yes- associated protein (Yes-associatedprotein, YAP) on proliferation and apoptosis of human renal cell carcinoma 786 cells. Methods: 786 cells of renal cell carcinoma were cultured in vitro. The shRNA lentivirus interference vector targeting YAP gene was constructed and transfected into 786 / 0 cells. The expression of YAP and its downstream transcription factor TEAD1mRNA and protein were detected by RT-PCR and Western blot before and after interfering with 786 / 0 cells, respectively. the expression of YAP and its downstream transcription factor TEAD1mRNA and protein were detected before and after interference. CCK-8 (cell counting kit-8) was used to detect the changes of cell proliferation after silencing YAP, and flow cytometry (flow cytometry, FCM) was used to detect the changes of apoptosis and cycle. Results: three YAP-shRNA lentivirus interference vectors targeting YAP gene and one negative control lentivirus vector were constructed and successfully transformed into 786 cells. After 96 hours of transfection with YAP-shRNA lentivirus interference vector, the expression of YAP mRNA and protein in 786 / 0 cells was significantly down-regulated (P 鈮

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