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目的探讨外源性野生型PTEN基因对人多发性骨髓瘤(MM)细胞株RPMI8226增殖、凋亡、侵袭活性的影响以及PTEN/FAK信号传导通路在细胞侵袭活性中的作用。方法将携带人野生型PTEN及绿色荧光蛋白的腺病毒(Ad-PTEN-GFP)及空载体腺病毒(Ad-GFP)体外转染至RPMI8226细胞。通过MTT法检测细胞生长曲线,Hoechst33342染色检测细胞凋亡,荧光定量PCR(FQ-PCR)检测PTEN及FAKmRNA水平变化,Western印迹检测PTEN、FAK及p-FAK蛋白变化,Transwell小室检测RPMI8226细胞侵袭能力。结果Ad-PTEN-GFP以感染复数为100转染RP-MI8226细胞后,第4天达到最大生长抑制率为42.01%,Ad-PTEN-GFP转染RPMI8226细胞72h后细胞凋亡率为(35.02±6.80)%,PTENmRNA及蛋白表达升高,FAKmRNA以及FAK、p-FAK蛋白表达下调,与Ad-GFP组相比差异显著(P<0.01,P<0.05);Transwell小室结果显示,转染Ad-PTEN-GFP组漏入下室内及黏附于下室面细胞数量较Ad-GFP组显著减少(P<0.01)。结论转染野生型PTEN基因的RPMI8226细胞增殖受抑,凋亡增加,细胞侵袭力明显减弱,其作用可能与下调FAK、p-FAK表达有关。
Objective To investigate the effect of exogenous wild-type PTEN gene on the proliferation, apoptosis and invasion of human multiple myeloma (MM) cell line RPMI8226 and the role of PTEN / FAK signaling pathway in cell invasion. Methods Ad-PTEN-GFP and Ad-GFP carrying human wild type PTEN and Ad-GFP were transfected into RPMI8226 cells in vitro. The cell growth curve was detected by MTT assay. Apoptosis was detected by Hoechst33342 staining. The changes of PTEN and FAK mRNA were detected by FQ-PCR. The protein expression of PTEN, FAK and p-FAK were detected by Western blotting. The invasion ability of RPMI8226 cells . Results The maximum growth inhibition rate of Ad-PTEN-GFP was 42.01% after transfection of RP-MI8226 cells with a multiplicity of infection of 100. The apoptosis rate of RPMI8226 cells transfected with Ad-PTEN-GFP was (35.02 ± (P <0.01, P <0.05). The results of Transwell chamber showed that the expression of PTEN mRNA and protein, the expression of FAK mRNA and FAK, p-FAK protein in Ad- Compared with Ad-GFP group, the number of PTEN-GFP group leaking into the lower chamber and adhering to the lower chamber was significantly decreased (P <0.01). CONCLUSION: The proliferation of RPMI8226 cells transfected with wild-type PTEN gene is inhibited, the apoptosis is increased, and the invasiveness of cells is obviously weakened. The effect may be related to the down-regulation of FAK and p-FAK expression.