摘要
目的:研究分析环孢素A对前列腺癌PC-3细胞紫杉醇耐药的作用及机制。方法:采用逐渐增量法建立PC-3耐药细胞株(PC-3R),分为空白对照组(未处理)、多西紫杉醇组(10-5 mol/L)及环孢素A低、中、高剂量组(10-5 mol/L多西紫杉醇+5μg/mL、10μg/mL、20μg/mL环孢素A),采用MTT法检测不同浓度多西紫杉醇对PC-3、PC-3R细胞株增殖抑制率以及不同剂量环孢素A联合紫杉醇对PC-3R细胞增殖抑制作用;采用流式细胞技术检测环孢素A联合紫杉醇对PC-3R细胞凋亡及细胞周期的影响;Western blotting法检测环孢素A联合紫杉醇对MTDH蛋白表达的影响。结果:随着多西紫杉醇浓度的增加,PC-3以及PC-3R细胞株增殖抑制率明显升高,且同一浓度下PC-3细胞株抑制率明显高于PC-3R(P<0.05)。多西紫杉醇组及环孢素A各剂量组细胞增殖抑制率及凋亡率均高于空白对照组,环孢素A低、中、高剂量组细胞增殖抑制率及凋亡率均高于多西紫杉醇组,并呈剂量依赖性(均P<0.05)。多西紫杉醇组及环孢素A各剂量组细胞G0/G1期比例较空白对照组显著升高,而S期显著降低(P<0.05);随着环孢素A药物浓度的增加,G0/G1期显著升高,S期比例显著降低(P<0.05);环孢素A低、中、高剂量组MTDH蛋白相对表达量较空白对照组及多西紫杉醇组明显降低,并呈剂量依赖性(P<0.05)。结论:环孢素A可促进前列腺癌耐药细胞株PC-3R化疗敏感性,其作用机制可能与促进细胞凋亡、阻滞细胞周期及下调MTDH蛋白表达有关。
Objective:To study the effect and mechanism of cyclosporine A on paclitaxel resistance in prostate cancer PC-3 cells.Methods:PC-3 drug resistant cell line(PC-3R)was established by gradual increment method.The cells were divided into blank control group(untreated),docetaxel group(10-5 mol/L)and cyclosporine A low,middle and high dose groups(10-5 mol/L docetaxel+5μg/mL,10μg/mL,20μg/mL cyclosporine A).MTT assay was used to detect the inhibitory rate of docetaxel at different concentrations on the proliferation of PC-3 and PC-3R cell lines,and the inhibitory effect of cyclosporine A at different concentrations combined with paclitaxel on the proliferation of PC-3R cells.Flow cytometry was used to detect the effect of cyclosporine A combined with paclitaxel on apoptosis and cell cycle of PC-3R cells.The effect of cyclosporine A combined with paclitaxel on MTDH protein expression was detected by Western blotting.Results:With the increase of docetaxel concentration,the proliferation inhibition rate of PC-3 and PC-3R cell lines increased significantly,and the inhibition rate of PC-3 cell line was significantly higher than that of PC-3R at the same concentration(P<0.05).The cell proliferation inhibition rate and apoptosis rate in docetaxel group and cyclosporine A low,middle and high dose groups were higher than those in blank control group,and the cell proliferation inhibition rate and apoptosis rate in low,middle and high dose groups were higher than those in docetaxel group,and in a dose-dependent manner(all P<0.05).The G0/G1 phase ratio of docetaxel group and cyclosporine A low,middle and high groups was significantly higher than that of the blank control group,while the S phase ratio was significantly decreased(P<0.05).With the increase of cyclosporine A concentration,the G0/G1 phase increased significantly,and the ration of S phase decreased significantly(P<0.05).The relative expression of MTDH protein in cyclosporine A low,middle and high dose groups was significantly lower than that in the blank control group and docetaxel group,and in a dosedependent manner(P<0.05).Conclusion:Cyclosporine A can promote the chemosensitivity of drug-resistant in prostate cancer PC-3R cell line,and its mechanism may be related to the promotion of apoptosis,cell cycle block and down-regulation of MTDH protein expression.
作者
李江涛
张静
Li Jiangtao;Zhang Jing(Department of Urinary Surgery,Geriatric Hospital of Hebei Province,Baoding 050000,China;Outpatient Department,Baoding No.2 Central Hospital,Baoding 050000,China)
出处
《广西医科大学学报》
CAS
2020年第10期1804-1809,共6页
Journal of Guangxi Medical University
基金
河北省卫生部医药卫生科技资助项目(No.13010520181155)。
