RNA干扰STMN1基因表达通过Notch信号通路诱导胃癌细胞凋亡及增强化疗敏感性的研究

RNA Interference with STMN1 Gene Expression Induces Apoptosis and Enhances Chemosensitivity in Gastric Cancer Cells via Notch Signaling Pathway

[目的]探讨RNA干扰微管不稳定性蛋白(STMN1)基因表达通过Notch信号通路对胃癌细胞凋亡及化疗敏感性的影响。[方法] RT-PCR及Western blotting分别检测人胃黏膜上皮细胞GES-1和人胃癌细胞MKN28、SGC7901、BGC823中STMN1 mRNA及蛋白表达;STMN1-siRNA转染BGC823细胞,同时设定对照组和阴性对照组,转染48h后RT-PCR及Western blotting分别检测STMN1 mRNA及蛋白表达;后续实验分为对照组、STMN1-siRNA组、顺铂组、STMN1-siRNA+顺铂组,细胞培养48h后,CCK8及流式细胞仪分别检测细胞增殖及凋亡情况;Western blotting检测活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、Notch1、Hes1蛋白表达。[结果] STMN1在MKN28、BGC823、SGC7901胃癌细胞中的mRNA及蛋白表达均显著高于在GES-1细胞中的表达(P<0.05),选择BGC823细胞作为研究对象;转染STMN1-siRNA后BGC823细胞中STMN1的mRNA及蛋白表达均显著低于对照组(P<0.05);STMN1-siRNA组、顺铂组、STMN1-siRNA+顺铂组OD值及Notch1、Hes1蛋白表达均显著低于对照组,细胞凋亡率及Cleaved caspase3蛋白表达均显著高于对照组(P<0.05),STMN1-siRNA+顺铂组OD值及Notch1、Hes1蛋白表达均显著低于STMN1-siRNA组和顺铂组,细胞凋亡率及Cleaved caspase3蛋白表达均显著高于STMN1-siRNA组和顺铂组(P<0.05)。[结论] RNA干扰STMN1表达可通过抑制Notch信号通路增强顺铂对胃癌细胞增殖的抑制及凋亡的促进作用。

[Objective] To investigate the effect of microtubule instability protein(STMN1) gene expression on apoptosis and chemosensitivity of gastric cancer cells and its relation to Notch signaling pathway. [Methods] The expression of STMN1 mRNA and protein in human gastric epithelial GES-1 cells and human gastric cancer MKN28,SGC7901 and BGC823 cells were detected by RT-PCR and Western blotting,respectively. STMN1-siRNA was transfected into BGC823 cells,and m RNA and protein expression of STMN1 were detected by RT-PCR and Western blotting 48 h after transfection. The gastric cancer BGC823 cells were divided into the control group,STMN1-siRNA group,cisplatin group and STMN1-siRNA +cisplatin group. The cell proliferation and apoptosis were detected by CCK8 and flow cytometry respectively;the expression of Cleaved caspase3,Notch1,Hes1 protein were detected by Western blotting. [Results] The expressions of STMN1 mRNA and protein in gastric cancer MKN28,BGC823 and SGC7901 cells were significantly higher than those in GES-1 cells(P<0.05). The mRNA and protein expressions of STMN1 in BGC823 cells were significantly lower than those in the control group after cells were transfected with STMN1-siRNA(P<0.05). The OD value,Notch1 and Hes1 protein expression in STMN1-siRNA group,cisplatin group and STMN1-siRNA+ cisplatin group were significantly lower than those in the control group,the apoptosis rate and the expression of Cleaved caspase3 protein were significantly higher than those in control group(P<0.05). The OD value,Notch1 and Hes1 protein ex-pression in STMN1-siRNA+ cisplatin group were significantly lower,and the apoptosis rate and the expression of Cleaved caspase3 protein were significantly higher than those in STMN1-siRNA group and cisplatin group. [Conclusion] RNA interference with STMN1 gene expression can inhibit the proliferation and promote the apoptosis of gastric cancer cells via Notch signaling pathway.