节律基因mClock在肿瘤实验化疗中的作用  

Function of Circadian Gene mClock on Tumor Experiment Chemotherapy

作　　者：要然[1] 段志青[1] 张太明[2] 甘露[1] 林丽[1] 刘延友[1] 汪宇辉[1] 王正荣[1] 

机构地区：[1]四川大学华西医学中心卫生部时间生物学重点实验室,四川成都610041 [2]四川大学华西医院胸心血管外科,四川成都610041

出　　处：《航天医学与医学工程》2008年第1期73-75,共3页Space Medicine & Medical Engineering

基　　金：国家自然科学基金资助项目(No.30470623;30470684)

摘　　要：目的研究节律基因mClock在顺铂诱导小鼠Lewis肺癌细胞(LLC)凋亡中的作用。方法体外采用12-十四酸佛波酯-13-乙酸盐(PMA)诱导LLC细胞近日节律,荧光定量聚合酶链式反应(PCR)检测细胞mClock基因的节律性表达。于诱导后不同时间点,顺铂(CDDP)处理细胞,24h后四甲基偶氮唑盐比色法(MTT)法和流式细胞术检测(FCM)其对细胞的增殖与凋亡的影响。采用脂质体介导方法将mClock基因转染至LLC细胞,顺铂处理细胞,24h后MTT法和流式细胞术检测其对细胞的增殖与凋亡的影响。结果PMA作用LLC后,mClock基因呈现节律性表达。PMA诱导LLC节律表达的不同时间点,细胞对顺铂的敏感性不同。mClock过表达使顺铂作用后LLC凋亡率下降,细胞增殖明显。结论mClock能够抑制顺铂诱导LLC的凋亡。Objective TO investigate the function of circadian gene mClock on the apoptosis of Lewis lung cancer （LLC） cells in mouse induced by cisplatin（CDDP）. Methods The circadian gene expression in LLC was induced with Phorbol 12-Myristate 13-acetate （PMA） in vitro and detected by realtime PCR, Lipofectamine-mediated gene transfection method was used to transfect mClock into LLC, After LLCs treated with PMA or transfected with mClock cisplatin was added to LLC at the indicated time. LLCs were incubated with cisplatin for 24 h, The cell proliferation and apoptosis were analyzed by MTT and FCM, respectively. Results After induced with PMA the expression of mClock gene was presented circadian pattern, whose peak and nadir were at 12 h and 24 h, respectively, The sensitivity of LLC to cisplatin was different at different time, at 24 h was higher than that at 12 h according to the results of MTT and FCM, LLC of circadian mClock gene over-expression revealed decreased rate of apoptosis and increased cell proliferation, Conclusion The circadian gene mClock may participate in the regulation of cisplatin reducing apoptosis.

关 键 词：近日节律 顺铂 mClock基因 LEWIS肺癌细胞 PMA 增殖 凋亡 

分 类 号：Q789[生物学—分子生物学]