青藤碱联合顺铂对顺铂耐药Hela细胞株增殖凋亡的作用

Effects of sinomenine combined with cisplatin on proliferation and apoptosis of cisplatin-resistant cell lines

目的探讨青藤碱联合顺铂对顺铂耐药Hela细胞株增殖凋亡的作用。方法取对数期Hela细胞,建立顺铂耐药模型,分为对照组、青藤碱组、激活剂组、青藤碱联合激活剂组。对照组不处理,青藤碱组加入青藤碱(40μmol/L终浓度),激活剂组加入Notch信号通路配体Jagged1蛋白(500 ng/ml),青藤碱联合激活剂组加入青藤碱(40μmol/L终浓度)+Jagged1蛋白(500 ng/ml)。MTT法检测不同浓度顺铂对各组细胞的增殖抑制率;流式细胞术检测细胞凋亡率;Western blot法检测细胞神经源性基因Notch同源蛋白1(Notch1)、信号转导及转录激活子3(STAT3)、发状分裂相关增强子1(Hes1)蛋白表达。结果与对照组比较,青藤碱组不同顺铂浓度对细胞增殖抑制率升高,IC_(50)降低,激活剂组不同顺铂浓度对细胞增殖抑制率降低,IC_(50)升高(P<0.05);与青藤碱组比较,青藤碱联合激活组不同顺铂浓度对细胞增殖抑制率降低,IC_(50)升高(P<0.05);与激活剂组比较,青藤碱联合激活组不同顺铂浓度对细胞增殖抑制率升高,IC_(50)降低(P<0.05);与对照组比较,青藤碱组凋亡率升高(P<0.05),激活剂组凋亡率降低(P<0.05);与青藤碱组比较,青藤碱联合激活组凋亡率降低(P<0.05);与激活剂组比较,青藤碱联合激活组凋亡率升高(P<0.05);与对照组比较,青藤碱组Notch1、Stat3、Hes1蛋白表达量降低(P<0.05),激活剂组Notch1、Stat3、Hes1蛋白表达量升高(P<0.05);与青藤碱组比较,青藤碱联合激活剂组Notch1、Stat3、Hes1蛋白表达量升高(P<0.05);与激活剂组比较,ATL联合转染组Notch1、Stat3、Hes1蛋白表达量降低(P<0.05)。结论青藤碱可提高化疗药物对宫颈癌细胞的增殖抑制率,并促进其凋亡,从而减弱宫颈癌细胞化疗耐药性,可能通过抑制Notch1/STAT3信号通路来实现。

Objective To explore the effect of sinomenine combined with cisplatin on proliferation and apoptosis of cisplatin-resistant cell lines.Methods Hela cells in the log phase were used to establish a cisplatin resistance model and divided into the control group,sinomenine group,activator group,and sinomenine combined with activator group.The control group was left untreated,sinomenine was added to the sinomenine group(40μmol/L final concentration),the Notch signaling pathway ligand Jagged1 protein(500 ng/ml)to the activator group,and sinomenine(40μmol/L final concentration)+Jagged1 protein(500 ng/ml)to the sinomenine combined with activator group.An MTT method was used to detect the proliferation-inhibiting rate of different concentrations of cisplatin in each group of cells.Flow cytometry was used to detect the rate of cell apoptosis.Western blot was used to detect the expressions of the cellular neurogenic gene Notch homologous protein 1(Notch1),signal transduction and Transcription activator 3(STAT3),and hair split-associated enhancer 1(Hes1)protein.Results Compared with the control group,different concentrations of cisplatin in the sinomenine group increased the proliferation-inhibiting rate of cells,but IC50 was decreased.In the activator group,different concentrations of cisplatin reduced the proliferation-inhibiting rate of cells,but IC50 was increased(P<0.05).Compared with the sinomenine group,different concentrations of cisplatin in the sinomenine combined with activator group reduced the proliferation-inhibiting rate of cells but increased IC50(P<0.05).Compared with the activator group,different concentrations of cisplatin in the sinomenine combined with activator group increased the inhibition rate of cell proliferation but decreased IC50(P<0.05).Compared with the control group,the apoptosis rate in the sinomenine group was increased(P<0.05),but decreased in the activator group(P<0.05).Compared with the sinomenine group,the apoptosis rate in the sinomenine combined with activator group was increased(P<0.05).Compared with the activator group,the apoptosis rate in the sinomenine combined with activator group was increased(P<0.05).Compared with the control group,the expressions of Notch1,Stat3 and Hes1 in the sinomenine group were decreased(P<0.05),but increased in the activator group(P<0.05).Compared with the sinomenine group,the expressions of Notch1,Stat3 and Hes1 protein in the sinomenine combined with activator group were increased(P<0.05).Conclusions Sinomenine can increase the proliferation-inhibiting rate of chemotherapeutic drugs on cervical cancer cells and promote their apoptosis,thereby reducing the chemotherapy resistance of cervical cancer cells.This is possibly due to the inhibition of the Notch1/STAT3 signaling pathway.