人耐紫杉醇去势性前列腺癌细胞系的构建  

Establishment of paclitaxel-resistant castration-resistant prostate cancer cells

作　　者：吴昌学[1] 李毅[1] 张婷[1] 彭煜晖 张健[1] 禹文峰[1] 柏华 张启芳[1] WU Changxue;LI Yi;ZHANG Ting;PENG Yuhui;ZHANG Jian;YU Wenfeng;BAI Hua;ZHANG Qifang(Education Ministry Key Laboratory of Endemic and Minority Diseases&Key Laboratory of Molecular Biology of Guizhou Medical University,Guiyang 550004,Guizhou,China;Medical Laboratory Center,the Third Affiliated Hospital of Guizhou Medical University,Duyun 558000,Guizhou,China)

机构地区：[1]贵州医科大学地方病与少数民族疾病教育部重点实验室&贵州省医学分子生物学重点实验室,贵州贵阳550004 [2]贵州医科大学第三附属医院医学实验中心,贵州都匀558000

出　　处：《贵州医科大学学报》2021年第6期621-626,共6页Journal of Guizhou Medical University

基　　金：国家自然科学基金(31960137);贵州省科技厅重点基金[黔科合基础(2016)1416];贵州省科技厅区域常见疾病与成体干细胞转化研究创新平台[黔科中引地(2019)4008];贵州省科技厅支撑计划[黔科合支撑(2020)4Y129]。

摘　　要：目的构建人耐紫杉醇(PTX)去势性前列腺癌细胞模型。方法培养人细胞株DU145细胞,逐渐增加PTX的浓度(1、4、6、8、10及12 nmol/L)处理细胞;以12 nmol/L浓度处理后存活的DU145细胞作为PTX耐药细胞株(命名为DU145 R)、DU145细胞作为对照,采用细胞增殖检测(CCK8)法检测DU145和DU145 R细胞的细胞活力、光学显微镜下观察细胞核形态、实时荧光定量PCR(qPCR)法检测细胞多药耐药基因1(MDR 1)和c-Myc mRNA表达水平、Western blot法检测细胞MDR1和c-Myc蛋白表达水平。结果与DU145细胞比较,显微镜下观察发现DU145 R细胞核里有多核化现象,DU145 R细胞活力明显高于DU145细胞(P<0.01),MDR1及c-Myc mRNA和蛋白表达水平明显高于DU145细胞(P<0.01)。结论成功构建DU145 R细胞,该细胞MDR 1及c-Myc表达水平上调,对PTX耐药。Objective To construct a paclitaxel-resistant castration-resistant prostate cancer(CRPC)cell model for investigating the molecular mechanism by which CRPC cells are resistant to paclitaxel(PTX).Method Parental DU145 cells were cultured with paclitaxel at gradually increased concentration of 1,4,6,8,10 and 12 nmol/L.The DU145 R cells survived after the treatment of 12 nmol/L concentration were chosen as the PTX resistant cell strain,and DU145 cell was chosen as control.CCK8 assay was used to compare cell viability of DU145 and DU145 R cells.Light microscope was used to observe nuclear features.Real-time fluorescent quantitative PCR(qPCR)was used to test mRNA expression level of multiple drug resistant gene 1(MDR 1)and c-Myc.Western blot was used to detect protein expression levels of MDR1 and c-Myc.Results Compared with DU145,DU145 R cells were observed to be multi-nucleated.The cell viability of DU145 R cells was higher than that of DU145 cells(P<0.01).Additionally,expression level of mRNA and protein in MDR1 and c-Myc were significantly higher than DU145 R cells(P<0.01).Conclusion Constructed DU145 R cells were resistant to PTX and its MDR1 and c-Myc expression levels were upregulated.

关 键 词：前列腺肿瘤 前列腺癌细胞 紫杉醇 耐药 多药耐药基因1 C-MYC 

分 类 号：R737.25[医药卫生—肿瘤]