CKMT1A对人鼻咽癌HONE1细胞铂类化疗药物敏感性的影响及机制研究  

Mitochondrial creatine kinase 1A in sensitivity of nasopharyngeal carcinoma cells to platinum-based chemotherapeutics

作　　者：李刚[1] LI Gang(Department of Ear,Nose and Throat,Wenchang People's Hospital,Wenchang 571300,Hainan,China)

机构地区：[1]文昌市人民医院耳鼻咽喉科

出　　处：《中南医学科学杂志》2019年第6期576-580,共5页Medical Science Journal of Central South China

摘　　要：近年来鼻咽癌患者五年生存率的提升难度明显增加,对铂类化疗药物的敏感性较差是主要原因,探讨耐药机制具有重要意义。本研究将鼻咽癌细胞转染分为空载体对照组(空载体质粒)、线粒体肌酸激酶1A(CKMT1A)过表达组(CKMT1A过表达载体质粒),分别进行MTT法检测、流式细胞术检测、实时荧光定量PCR检测和Western blot检测。结果显示随着顺铂浓度的增加,CKMT1A过表达组吸光度值下降幅度更明显(P<0.01)。根据顺铂IC 50结果选取2μmol/L顺铂处理细胞24 h后,两组细胞均表现为G0/G1期比例明显下降,S期和G2/M期比例明显增多。2μmol/L顺铂处理细胞48 h后,CKMT1A过表达组细胞凋亡率明显高于空载体对照组(P<0.01);CKMT1A过表达组c-PARP、Bax相对表达量明显高于空载体对照组,BCL-2相对表达量明显低于空载体对照组(P<0.05)。2μmol/L顺铂处理细胞6 h后,CKMT1A过表达组p-STAT3相对表达量明显高于空载体对照组(P<0.01)。过表达CKMT1A可通过细胞凋亡信号通路下调STAT3磷酸化水平,抑制靶基因表达,促进人鼻咽癌细胞株HONE1凋亡,提升对铂类化疗药物的敏感性。In recent years,it is more difficult to improve the five-year survival rate of NPC patients.The poor sensitivity to platinum chemotherapy drugs is the main reason.It is of great significance to explore the mechanism of drug resistance.Cell transfection was divided into empty vector control group(empty vector plasmid)and CKMT1A overexpression group(CKMT1Aoverexpression vector plasmid),detect by MTT method,flow cytometry,real-time fluorescence quantitative PCR and Western blot.The results showed that the absorbance values of both groups decreased,especially in CKMT1A overexpression group.According to the results of cisplatin IC 50,after 24 hours of treatment with 2μmol/L cisplatin,the proportion of G0/G1 phase decreased significantly in both groups,while the proportion of S phase and G2/M phase increased significantly.After 48 hours of treatment with 2μmol/L cisplatin,the apoptotic rate of CKMT1A overexpression group was significantly higher than that of empty vector control group(P<0.01);the relative expressions of c-PARP and Bax in both groups were significantly increased,while the relative expressions of BCL-2 were significantly decreased(P<0.05);the relative expressions of c-PARP and Bax in CKMT1A overexpression group were significantly higher than those in empty vector control group(P<0.05).The relative expression of BCL-2 was significantly lower than that of empty vector control group(P<0.05).After 6 hours of treatment with 2μmol/L cisplatin,the relative expression of p-STAT3 in CKMT1A overexpression group was significantly higher than that in empty vector control group(P<0.01).Overexpression of CKMT1A can down-regulate p-STAT3 level,inhibit target gene expression,promote apoptosis of human nasopharyngeal carcinoma cell line HONE1,and enhance the sensitivity to platinum chemotherapeutic drugs through related molecular mechanisms.

关 键 词：鼻咽癌 线粒体肌酸激酶1A 铂类化疗药物 

分 类 号：R739.6[医药卫生—肿瘤]