siRNA沉默TGFβR1对人口腔上皮癌Ca9-22细胞增殖和凋亡影响

The Effect of TGFβR1 Silencing by siRNA on Proliferation and Apoptosis of Human Oral Epithelial Carcinoma Ca9-22 Cells

为了研究转化生长因子β受体1 (transforming growth factor-βreceptor 1,TGFβR1)在人口腔上皮癌Ca9-22细胞中的表达,并探讨TGFβR1 siRNA对Ca9-22细胞增殖和凋亡的影响及分子机制,本研究通过荧光定量PCR检测TGFβR1 mRNA在人口腔上皮癌Ca9-22细胞中的表达水平。通过lipofectamine 2000将TGFβR1 siRNA转染到Ca9-22细胞,荧光定量PCR检测TGFβR1 siRNA干扰效率,CCK8和流式细胞术分别检测TGFβR1 siRNA对Ca9-22细胞增殖和凋亡的影响;Western blotting检测TGFβR1 siRNA对Ca9-22细胞Hes1、Bcl2和Notch1蛋白表达的影响。荧光定量PCR检测结果显示,与正常人口腔上皮HOK细胞比较,人口腔上皮Ca9-22细胞中TGFβR1 mRNA水平显著升高;CCK8结果显示,与Control组比较,TGFβR1siRNA组细胞增殖率显著降低(p<0.01);流式检测结果显示,与Control组比较,TGFβR1 siRNA组Ca9-22细胞凋亡率为显著升高(p<0.01);Western blotting结果显示,与Control组比较,TGFβR1 siRNA组Hes1、Bcl2和Notch1蛋白表达显著降低(p<0.01)。研究结果初步表示,siRNA干扰TGFβR1能够抑制Notch1/Hes1通路,抑制Ca9-22细胞增殖,促进Ca9-22细胞凋亡。

To study the expression of transforming growth factor β receptor 1(TGFβR1) in human oral epithelial carcinoma Ca9-22 cells and investigate the effect of TGFβR1 siRNA on proliferation and apoptosis and of Ca9-22 cells,the expression of TGFβR1 mRNA in Ca9-22 cells were detected by qRT-PCR.TGFβR1 siRNA was transfected into Ca9-22 cells by lipofectamine 2000.The effects of TGFβR1 siRNA on the proliferation and apoptosis of Ca9-22 cells were detected by CCK8 and flow cytometry.Western blotting was used to detect the expression of Hesl,Bcl2 and Notchl proteins in Ca9-22 cells transfected with TGFβR1 siRNA.The results of qRT-PCR showed that the expression of TGFβR1 mRNA in Ca9-22 cells was significantly higher than that in normal human oral epithelial HOK cells.The results of CCK8 showed that the proliferation rate of Ca9-22 cells in TGFβR1 siRNA transfection group was significantly lower than that in the control group.The results of flow cytometry showed that the apoptosis rate of Ca9-22 cells in the TGFβR1 siRNA group was significantly higher than that of the control group(p <0.01).The results of Western blotting showed that the expression of Hesl,Bcl2 and Notch1 protein in the TGFβR1 siRNA group was significantly higher than that of the control group(p<0.01).The results of this research presented that interference with TGFβR1 by siRNA can inhibit the Notchl pathway and the proliferation of Ca9-22 cells,and promote the apoptosis of Ca9-22 cells.