PRRX1对乳腺癌多药耐药发生的影响及其机制  

EFFECT OF PRRX1 ON MULTIDRUG RESISTANCE IN BREAST CANCER AND ITS MECHANISM

作　　者：罗皓月 董娇娇 姜丹丹[1] 王新刚[1] 陈庆峰[1] 李福年[1] LUO Haoyue;DONG Jiaojiao;JIANG Dandan;WANG Xingang;CHEN Qingfeng;LI Funian(Breast Disease Diagnosis, Treatment Centre, The Affi-liated Hospital of Qingdao University, Qingdao 266003, China)

机构地区：[1]青岛大学附属医院乳腺病诊疗中心,山东青岛266003

出　　处：《精准医学杂志》2020年第3期213-216,221,共5页Journal of Precision Medicine

基　　金：国家自然科学基金资助项目(81302290)。

摘　　要：目的探讨配对相关同源框1(PRRX1)对乳腺癌多药耐药(MDR)发生的影响及其机制。方法选取乳腺癌MCF-7细胞进行培养,当细胞融合达80%后分离接种于24孔板中,细胞分为3组,未处理的MCF-7细胞为空白对照组(A组),转染空白载体的MCF-7细胞为阴性对照组(B组),转染携带PRRX1基因质粒的MCF-7细胞为实验组(C组)。用倒置荧光显微镜观察各组细胞形态变化及转染情况。通过RT-qPCR和WES蛋白印迹定量分析系统分别检测各组细胞PRRX1、N-钙黏蛋白(N-cadherin)和波形蛋白(vimentin)以及P-糖蛋白(P-gp)mRNA及蛋白表达水平。用CCK8试剂盒评估多西他赛和顺铂对各组细胞的半数抑制浓度(IC50)。结果B组和C组细胞均发出绿色荧光,转染效率约80%。PRRX1过表达使C组的MCF-7细胞由“铺路石”形态向“长梭形”形态转变。C组细胞的PRRX1、N-cadherin、vimentin和P-gp mRNA及蛋白表达水平与A、B组比较均明显升高(t=3.02~10.92,P<0.05)。经多西他赛和顺铂处理细胞24 h后,C组两种药物的IC50值明显高于A、B组(t=4.54~5.68,P<0.05)。结论PRRX1过表达可使乳腺癌MCF-7细胞发生MDR,其机制可能是通过诱导MCF-7细胞发生上皮间质转化而实现的。Objective To investigate the effect of paired related homeobox 1(PRRX1)on multidrug resistance(MDR)in breast cancer and the underlying mechanism.Methods Breast cancer MCF-7 cells were cultured,and then isolated and inoculated into 24-well plates after reaching 80%confluence.The untreated MCF-7 cells were used as blank control group(group A),the MCF-7 cells transfected with blank vector were used as negative control group(group B),and the MCF-7 cells transfected with the plasmid containing PRRX1 were used as experimental group(group C).The morphological changes and transfection of these cells were observed under an inverted fluorescence microscope.The mRNA and protein expression levels of PRRX1,N-cadherin,vimentin,and P-glycoprotein(P-gp)in these cells were measured by RT-qPCR and Western blot analysis,respectively.The half-maximal inhibitory concentrations(IC50)of docetaxel and cisplatin for these cells were evaluated by CCK8 assay.\ResultsGreen fluorescence was emitted from the cells in groups B and C,with a transfection efficiency of about 80%.In group C,overexpression of PRRX1 changed the morphology of MCF-7 cells from“paving stone”shape to“long spindle”shape.The mRNA and protein expression levels of PRRX1,N-cadherin,vimentin,and P-gp were significantly higher in group C than in groups A and B(t=3.02-10.92,P<0.05).Group C had a significantly higher IC50 of docetaxel and cisplatin,which were used to treat the MCF-7 cells for 24 h,than groups A and B(t=4.54-5.68,P<0.05).Conclusion Overexpression of PRRX1 may promote multidrug resistance in breast cancer MCF-7 cells,possibly by inducing epithelial-mesenchymal transition in these cells.

关 键 词：乳腺肿瘤 配对相关同源框1 N-钙黏蛋白 上皮-间充质转化 波形蛋白 P-糖蛋白 抗药性 肿瘤 体外研究 

分 类 号：R737.9[医药卫生—肿瘤]