SATB2基因siRNA对口腔鳞状细胞癌生长及STAT3信号的抑制作用

Inhibitory effect of SATB2 gene siRNA on the growth and STAT3 signal of oral squamous cell carcinoma

目的:探讨抑制SATB2基因表达对口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)细胞增殖、凋亡、侵袭和迁移的影响及机制。方法:以Lipofectamin^TM2000为载体,将阴性对照siRNA(阴性组)与SATB2特异性siRNA(抑制组)转染人OSCC细胞株CAL-27,设置空白组,CCK-8法检测siRNA转染4d的细胞增殖;siRNA转染CAL-27细胞48h,Western印迹法检测E-cadherin,STAT3,p-STAT3和cleaved caspase-3蛋白表达。克隆形成实验、Transwell小室及流式细胞术分别检测细胞克隆形成率、细胞侵袭和迁移能力及细胞凋亡率。结果:转染siRNA的CAL-27细胞SATB2蛋白表达明显低于空白组(P<0.05)。与空白组相比,抑制组细胞增殖、侵袭和迁移能力均明显降低,细胞凋亡率明显升高,E-cadherin和cleaved caspase-3蛋白表达明显升高,p-STAT3蛋白表达明显降低(P<0.05)。结论:RNA干扰抑制SATB2基因表达可抑制OSCC细胞生长,机制与下调STAT3信号有关。

Objective:To investigate the effect of inhibiting SATB2 gene expression on proliferation,apoptosis,invasion,and migration of oral squamous cell carcinoma cells and its mechanism.Methods:Lipofectamin^TM 2000 was used as vector,and negative control siRNA (negative group) and SATB2 specific siRNA group (inhibitory group) were transfected into human oral squamous cell carcinoma cell line CAL-27,and a blank group was set up,CCK-8 method was used to detect the proliferation of cells after siRNA was transfected into CAL-27 cells for 4 days,Western blotting were used to detect E-cadherin,STAT3,p-STAT3,and cleaved caspase-3 protein expression after siRNA was transfected into CAL-27 cells for 48 hours.Clone formation assay,Transwell chamber and flow cytometry were used to detect the cell clone formation rate,cell invasion and migration ability,and cell apoptosis rate.Results:The expression of SATB2 protein in CAL-27 cells transfected with siRNA was significantly lower compared with the blank group (P<0.05).Compared with the control group,the proliferation,invasion and migration of the cells in the inhibitory group decreased significantly,the apoptosis rate increased significantly,the expression of E-cadherin and cleaved caspase-3 protein increased significantly,and the expression of p-STAT3 protein was decreased significantly (P<0.05).Conclusion:Inhibition of SATB2 gene expression by RNA interference may inhibit the growth of oral squamous cell carcinoma cells,and the mechanism is related to down-regulation of STAT3 signal.