摘要
目的:研究葛根素联合γ突触核蛋白(synuclein gamma,SNCG)siRNA对膀胱癌细胞生长的影响。方法:用0,100,200,400,800,1 600μg/mL的葛根素培养液培养膀胱癌细胞,以MTT法检测细胞增殖变化并计算半数抑制浓度。在膀胱癌细胞中转染SNCG siRNA重组慢病毒和阴性对照慢病毒,以实时荧光定量PCR(quantitative real-time polymerase chain reaction,qRT-PCR)和Western印迹法检测干扰效果。用半数抑制浓度的葛根素处理稳定转染SNCG siRNA重组慢病毒的膀胱癌细胞,MTT法检测细胞增殖变化,平板克隆试验检测细胞克隆形成能力,Annexin V-FITC/PI双染法检测细胞凋亡变化,Western印迹法检测细胞中活化的caspase-3(C-caspase-3)、活化的caspase-9(Ccaspase-9)、Bcl-2相关X蛋白(Bax)蛋白表达水平。结果:100,200,400,800,1600μg/mL的葛根素处理后的膀胱癌细胞存活率均低于0μg/mL处理组(P<0.05),其半数抑制浓度为(587.26±61.45)μg/mL。转染SNCGsiRNA重组慢病毒的膀胱癌细胞中SNCG表达水平低于转染阴性对照重组慢病毒的膀胱癌细胞(P<0.05)。下调SNCG或葛根素处理后的膀胱癌细胞增殖能力和克隆形成能力均降低,细胞凋亡率升高,细胞中C-caspase-3,C-caspase-9,Bax蛋白水平也升高。与下调SNCG或葛根素处理后的膀胱癌细胞比较,葛根素处理下调SNCG的膀胱癌细胞的增殖和克隆形成能力均降低,细胞凋亡率升高,细胞中C-caspase-3,C-caspase-9,Bax蛋白表达水平也升高。结论:葛根素联合SNCG siRNA能够抑制膀胱癌细胞生长、诱导膀胱癌细胞凋亡。
Objective: To study the effect of puerarin combined with synuclein gamma (SNCG) siRNA on the growth of bladder cancer cells. Methods: Bladder cancer cells were cultured with 0, 100, 200, 400, 800 and 1 600 μg/mL of Puerarin culture medium. Cell proliferation was measured by MTT and the median inhibitory concentration was calculated. SNCG siRNA was transfected into bladder cancer cells by recombinant lentivirus and negative control recombinant lentivirus, the jamming effect was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. The bladder cancer cells stably transfected with SNCG siRNA recombinant lentivirus were treated with Puerarin at half inhibitory concentration. The proliferation of the cells was detected by MTT assay, the colony forming ability of cells was detected by plate cloning test, the apoptosis was uesd by Annexin V-FITC/PI double staining, the expression levels of C-caspase-3, C-caspase-9 and Bax in the cells were uesd by Western blot. Results: The survival rate of bladder cancer cells treated with 100, 200, 400, 800 and 1 600 μg/mL puerarin was lower than that treated with 0 μg/mL (P<0.05), the half inhibitory concentration was calculated to be (587.26±61.45)μg/mL. The expression level of SNCG in bladder cancer cells transfected with SNCG siRNA recombinant lentivirus was lower than that in bladder cancer cells transfected with negative control recombinant lentivirus (P<0.05). The cell proliferation and colony formation ability of bladder cancer cells treated with SNCG or puerarin decreased, the rate of apoptosis increased, the level of C-caspase-3, C-caspase-9 and Bax protein also increased. Compared with bladder cancer cells treated with SNCG or puerarin, puerarin decreased the proliferation and colony formation ability of bladder cancer cells which downregulated SNCG. The rate of apoptosis increased, the level of C-caspase-3, C-caspase-9 and Bax protein also increased. Conclusion: Puerarin combined with SNCG siRNA can inhibit bladder cancer cell growth
作者
王国政
沈金贵
邹永平
汪雪
李笑蕾
WANG Guozheng;SHEN Jingui;ZOU Yongping;WANG Xue;LI Xiaolei(Department of Urology, Shanghai Xuhui Dahua Hospital, Shanghai 200237;Department of Neurology, China-Japan Friendship Hospital, Jilin University, Changchun 130033, China)
出处
《临床与病理杂志》
2018年第12期2558-2565,共8页
Journal of Clinical and Pathological Research
