葛根素联合γ突触核蛋白siRNA对膀胱癌细胞生长的抑制作用
| 作者: |
1王国政,
1沈金贵,
1邹永平,
1汪雪,
2李笑蕾
1 上海市徐汇大华医院泌尿外科，上海 200237 2 吉林大学中日联谊医院神经内科，长春 130033 |
| 通讯: |
李笑蕾
Email: yongxing877134@163.com |
| DOI: | 10.3978/j.issn.2095-6959. 2018.12.005 |
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0，100，200，400，800，1 600 μg/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(C-caspase-9)、Bcl-2相关X蛋白(Bax)蛋白表达水平。结果：100，200，400，800，1 600 μg/mL的葛根素处理后的膀胱癌细胞存活率均低于0 μg/mL处理组(P<0.05)，其半数抑制浓度为(587.26±61.45) μg/mL。转染SNCG siRNA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。
关键词：
膀胱癌；生长；γ突触核蛋白；葛根素
Inhibitory effect of puerarin combined with SNCG siRNA on bladder cancer cell growth
CorrespondingAuthor: LI Xiaolei Email: yongxing877134@163.com
DOI: 10.3978/j.issn.2095-6959. 2018.12.005
Abstract
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 and induce bladder cancer cell apoptosis.
Keywords：
bladder cancer; growth; synuclein gamma; puerarin