苦参碱及其衍生物对人鼻咽癌细胞的耐药逆转作用

Reversal Effect of Matrine on Drug Resistance of Human Nasopharyngeal Carcinoma Cell Line

目的:探讨中药苦参碱(matrine,MAT)及其衍生物对鼻咽癌耐药细胞HONE1/DDP的耐药逆转作用。方法:采用逐渐增加剂量的方法,诱导建立人鼻咽癌细胞HONE1耐顺铂细胞株HONE1/DDP;将不同浓度苦参碱及其衍生物按倍比稀释浓度分别加入到HONE1/DDP中,测定其细胞毒性;MTT法测定非细胞毒性浓度苦参碱及其衍生物对HONE1/DDP细胞株的逆转作用,流式细胞仪检测细胞周期分布情况,并以Western blot测定MRP1、BAX、BCL-2蛋白表达。结果:HONE1/DDP细胞系对顺铂耐药;经苦参碱、苦参碱衍生物作用24 h后,HONE1/DDP细胞株对顺铂的耐药性下降,逆转指数分别为1.45、1.77;与HONE1/DDP比较,经非细胞毒性浓度苦参碱处理的HONE1/DDP细胞周期G0/G1明显增加(P<0.05),经非细胞毒性浓度苦参碱衍生物处理的HONE1/DDP细胞周期S期减少(P<0.05);HONE1/DDP细胞的MRP1表达显著高于HONE1(P<0.05),而经苦参碱、苦参碱衍生物作用后MRP1表达显著降低(P<0.05),且二者作用后BAX表达水平降低,BCL-2表达水平升高,BAX/BCL-2比值降低(P<0.05)。结论:人鼻咽癌细胞株HONE1对顺铂耐药,其耐药机制可能与增加MRP1的表达有关;苦参碱及其衍生物可以逆转HONE1/DDP对顺铂的耐药性,其机制可能与其改变细胞周期分布及下调MRP1的表达、降低BAX/BCL-2比值有关。

Objective：To study the reversal effect of matrine and its derivatives on drug resistance of human nasopharyngeal carcinoma cell line HONE1/DDP.Methods：The drug-resistant cell line was established by the human nasopharyngeal carcinoma cells HONE1 with gradually increasing concentration of cisplatin.The matrine and its derivatives were added into the HONE1/DDP according to different concentrations to measure their cytotoxicity.MTT assay was used to measure the reversal effect on the drug resistance of the non-toxic doses of matrine and its derivatives.Cell cycle was measured by flow cytomety.The expression level of MRP1,BAX,BCL-2 was assayed by Western blot.Results：HONE1/DDP indicated drug resistantance.When the non-toxic doses of matrine and its derivatives were used to HONE1/DDP with 24h,the resistance of HONE1/DDP was down-regulated,the reversal fold was 1.45 and 1.77.The cell number of G0/G1-phase increased in matrine group while S phase decreased in matrine derivatives group compared with HONE1/DDP.Compared with HONE1/DDP group,the MRP1 expression levels in HONE1 cells were reduced（P〈0.05）,and Matrine group and matrine derivatives group were enhanced（P〈0.05）.The expression of BAX was lower while the expression of BCL-2 was higher.Conclusion：The resistance of HONE1 cells to DDP is able to increase the expression of MRP1;Matrine and its derivatives can reverse the drug resistance of HONE1/DDP to DDP,its activity may be related to change cell cycle distribution,the inhibition of MRP1 expression and down-regulation of BAX/BCL-2.