HPA-siRNA促进皮肤鳞癌A431细胞凋亡并抑制其增殖、侵袭和迁移的机制

目的探讨乙酰肝素酶(HPA)小干扰RNA(siRNA)对皮肤鳞状细胞癌A431细胞增殖、凋亡、侵袭和迁移的影响及其机制。方法将体外培养的A431细胞分为空白对照组(未转染)、阴性对照组(转染NC-siRNA)和HPA沉默组(转染HPA-siRNA),RT-PCR检测各组细胞中HPA mRNA的表达,MTT法、流式细胞仪和Transwell分别检测细胞增殖、凋亡、侵袭和迁移;Western blot检测细胞中HPA蛋白和PI3K/AKT信号通路相关蛋白PCNA、MMP-2、Bcl-2、p-AKT和AKT的表达。结果与空白对照组相比,HPA沉默组细胞在2~3 d生长过程中的A值明显下降,侵袭和迁移细胞数均明显减少,HPA mRNA和HPA、PCNA、MMP-2、Bcl-2、p-AKT蛋白的表达水平均明显降低,细胞凋亡率明显升高(P<0.05);而空白对照组与阴性对照组间差异无统计学意义(P>0.05)。结论HPA siRNA可抑制A431细胞增殖、侵袭和迁移并诱导细胞凋亡,其作用机制可能与抑制PI3K/AKT信号通路的活化有关。

HPA-siRNA Promotes Apoptosis and Inhibits Proliferation,Invasion and Migration of Skin Squamous Cell Carcinoma A431 Cells

Objective To investigate the effect and mechanism of small interfering RNA (siRNA) of heparanase (HPA) on proliferation,apoptosis,invasion and migration of cutaneous squamous cell carcinoma A431 cells. Methods A431 cells were divided into control group (untransfected),negative control group (transfected with NC-siRNA) and HPA silent group (transfected with HPA-siRNA).The expression of HPA mRNA was detected by RT-PCR,the proliferation,apoptosis,invasion and migration of cells were detected by MTT,flow cytometry and transwell respectively.The expression of HPA protein and PI3K/AKT signaling pathway related proteins PCNA,MMP-2,Bcl-2,p-AKT and AKT were detected by Western blot.Results Compared with the control group,the A value of cells was decreased significantly during 2~3 days of growth,the number of invasive and migrating cells were reduced significantly,the expression levels of HPA mRNA and HPA,PCNA,MMP-2,Bcl-2,p-AKT proteins were down-regulated significantly,and the apoptotic rate was increased significantly in HPA silent group (P<0.05).But there were no significant differences between negative control group and control group (P>0.05).Conclusion HPA siRNA can inhibit the proliferation,invasion and migration of A431 cells and induce apoptosis.The mechanism may be related to the inhibition of PI3K/AKT signaling pathway.