摘要
目的探讨沉默stathmin对顺铂(DDP)耐药鼻咽癌细胞耐药性的影响及其作用机制。方法采用浓度递增法建立DDP耐药鼻咽癌细胞CNE2/DDP,将si-stathmin、si-NC分别转染至CNE2/DDP细胞,并分别作为sistathmin组和si-NC组。采用不同浓度DDP处理CNE2、CNE2/DDP细胞及si-stathmin组、si-NC组CNE2/DDP细胞。噻唑蓝(MTT)法检测细胞的增殖抑制率;蛋白质印迹法(Western blot)检测细胞中stathmin、cleaved caspase 3、Bcl-2、SHH、PTCH1、GLI-1蛋白的表达水平;流式细胞术检测细胞的凋亡情况。结果DDP对CNE2细胞的半抑制浓度(IC50)为(0.243±0.012)μg/ml,对CNE2/DDP细胞的IC50为(1.347±0.070)μg/ml,耐药指数(RI)为5.591。DDP对si-stathmin组CNE2/DDP细胞的IC50为(0.582±0.025)μg/ml,对si-NC组CNE2/DDP细胞的IC50为(1.320±0.058)μg/ml。CNE2/DDP细胞中stathmin蛋白的相对表达量明显高于CNE2细胞(P﹤0.01);同一浓度DDP处理后,si-stathmin组CNE2/DPP细胞的增殖抑制率、凋亡率均明显高于si-NC组(P﹤0.01);si-stathmin组CNE2/DDP细胞中stathmin、Bcl-2、SHH、PTCH1、GLI-1蛋白的相对表达量均明显低于si-NC组,cleaved caspase 3蛋白的相对表达量明显高于si-NC组,差异均有统计学意义(P﹤0.01)。结论沉默stathmin可以逆转CNE2/DDP细胞的DDP耐药性,其机制可能与促进细胞凋亡,失活SHH信号通路有关。
Objective To study the effect of silencing stathmin on the drug resistance of cisplatin(DDP)-resistant nasopharyngeal carcinoma cells,and explore its mechanism.Method DDP-resistant nasopharyngeal carcinoma cells CNE2/DDP were established by increasing concentration method.si-stathmin and si-NC were transfected into CNE2/DDP cells as si-stathmin group and si-NC group,respectively.CNE2,CNE2/DDP cells and CNE2/DDP cells in si-stathmin group and si-NC group were treated with DDP at different concentrations.The inhibition rate of cell proliferation was detected by methylthiazolyldiphenyl-tertrazolium bromide(MTT)assay;the expression of stathmin,cleaved caspase 3,Bcl-2,SHH,PTCH1 and GLI-1 proteins in cells were detected by Western blot;apoptosis was detected by flow cytometry.Result The half maximal inhibitory concentration(IC50)of DDP on CNE2 cells was(0.243±0.012)μg/ml,and the IC50 of DDP on CNE2/DDP cells was(1.347±0.070)μg/ml,and the resistance index(RI)was 5.591.The IC50 of DDP on CNE2/DDP cells in si-stathmin group was(0.582±0.025)μg/ml,and the IC50 of DDP on CNE2/DDP cells in si-NC group was(1.320±0.058)μg/ml.The relative expression of stathmin protein in CNE2/DDP cells was significantly higher than that in CNE2 cells(P<0.01).After DDP treatment at the same concentration,the inhibition rate of proliferation and apoptosis rate of CNE2/DPP cells in si-stathmin group were significantly higher than those in si-NC group(P<0.01).The relative expression levels of stathmin,Bcl-2,SHH,PTCH1,and GLI-1 in CNE2/DDP cells of the si-stathmin group were significantly lower than those of the si-NC group,while that of cleaved caspase 3 protein was significantly higher compared to si-NC group,with statistically significant differences observed(P<0.01).Conclusion Silencing stathmin could reverse the resistance of CNE2/DDP cells to cisplatin.The underlying mechanism may be related to promoting apoptosis and inactivating SHH signaling pathway.
作者
汪文利
许慧
蒋树林
WANG Wenli;XU Hui;JIANG Shulin(Department of Otolaryngology,Kaifeng People’s Hospital,Kaifeng 475002,He’nan,China;Department of Medicine,PLA 73101 Army Hospital,Xuzhou 221008,Jiangsu,China)
出处
《癌症进展》
2019年第16期1894-1898,共5页
Oncology Progress
