甲基莲心碱对Tca8113/CBP卡铂耐药性的逆转作用  

Neferine reverses resistance to carboplatin in Tca8113/CBP cells

作　　者：王启明[1] 李勇[1] 李显[1] 童熹[1] 李婷[1] 

机构地区：[1]重庆医科大学附属口腔医院口腔颌面外科,重庆400015

出　　处：《第三军医大学学报》2011年第15期1615-1619,共5页Journal of Third Military Medical University

摘　　要：目的建立人舌鳞癌卡铂耐药细胞株Tca8113/CBP,探讨甲基莲心碱Nef对Tca8113/CBP卡铂耐药性的逆转作用及其机制。方法采用逐步递增卡铂浓度、体外间歇诱导法建立Tca8113/CBP耐卡铂细胞株;MTT法检测Nef对Tca8113/CBP卡铂耐药性的逆转作用;RT-PCR法检测Nef作用前后Tca8113/CBP内耐药相关基因肺耐药蛋白LRP与成纤维细胞肌型原肌球蛋白FMT的表达水平;免疫荧光观察Nef作用前后细胞形态结构变化。结果历时5个月建立的Tca8113/CBP细胞株对卡铂耐药性稳定,耐药指数为4.06。Nef对Tca8113/CBP的卡铂耐药性有逆转作用,逆转倍数达到2.12倍;与Tca8113/CBP细胞比较,经Nef干预后,Tca8113/CBP细胞内LRP与FMT mRNA表达水平明显降低(P<0.01),细胞内F-actin的含量也明显下降,排列整齐,分布趋于均匀,接近非耐药Tca8113细胞水平。结论 Nef对Tca8113/CBP的卡铂耐药性有逆转作用,其机制可能与干预Tca8113/CBP细胞内LRP和FMT表达,增加Tca8113/CBP细胞内卡铂的含量,改变微丝结构,降低细胞黏附能力,干扰细胞增殖周期有关。Objective To establish a carboplatin （CBP）-resistant tongue squamous carcinoma cell line, TcaSll3/CBP cells, and to observe the reversal effect of neferine in these cells. Methods TcaSll3/ CBP cell line was established by gradually increasing the concentration of carboplatin and intermittent inducement to Tca8113 ceils. Methyl thiazolyl tetrazolium （MTT） assay was used to evaluate the reversal effect of neferine on proliferation in Tca8113/CBP cells, mRNA expression of drug-resistance-associated genes, lung related resistant protein （LRP） and fibroblast muscle-type tropomyosin （FMT） were monitored by RT-PCR in the cells before and after resistance establishment. Morphology of the cells was observed under fluorescent microscopy, and the intracellular changes of cytoskeleton between reversed Tca8113/CBP and non-reversed Tca8113/CBP were compared. Results After 5 month＇ s CBP treatment, Tca8113/CBP cells which can be stably resistant to carboplatin （16 μg/ml） was successfully established. The resistance index is 4.06. Neferine reversed the resistance of Tca8113/CBP cells with a reversal fold of 2.12. Compared to the Tca8113 ceils, the expression of LRP and FMT at mRNA level was increased obviously in Tca8113/CBP ceils. Whereas they were decreased after neferine treatment （P 〈 0.01 ）, and the intracellular F-actin content was also decreased, arranged in neat rows, more evenly distributed close to the level of non-resistant Tca8113 cells. Conclusion Neferine exhibits the reversal effect on Tca8113/CBP cells, which may be relate to decrease of LRP and FMT expression, increase of intracellular content of carboplatin, change of the microfilament, reduce of mechanical properties of adhesion, and interference of cell cycle in Tca8113/CBP cells.

关 键 词：甲基莲心碱 Tca8113/CBP 卡铂 耐药性 逆转 成纤维细胞肌型原肌球蛋白 

分 类 号：R285.5[医药卫生—中药学] R739.86[医药卫生—中医学]