摘要
目的:探讨丹参酮IIA对阿霉素(又称多柔比星,DOX)耐药胃癌细胞的抑制作用及具体机制。方法:采用MTT法检测胃癌细胞对DOX的敏感性;逐步筛选得到耐DOX细胞株,流式细胞术及Western blot检测细胞周期、凋亡及自噬相关的标志物;RT-qPCR及Western blot检测渗透性糖蛋白(P-gp)、乳腺癌耐药蛋白(BCRP)及多重耐药相关蛋白1(MRP-1)的表达。结果:根据DOX对不同细胞的IC 50,鉴定出DOX敏感细胞株SNU-719和SNU-601及DOX耐药细胞株SNU-638、SNU-668、SNU-216和SNU-620。同时筛选得到两个耐药株SNU-719R和SNU-601R。丹参酮IIA抑制耐DOX细胞的MRP-1表达。与单用DOX处理组相比,丹参酮IIA与DOX联用能减少SNU-719R及SNU-620细胞的G 2/M期细胞,增加p21表达水平,降低cyclin B1及CDK1的表达水平。另外,与单用DOX处理组相比,丹参酮IIA与DOX联用能增加p53、Bax和LC3B-II的表达水平,降低Bcl-2和p62的水平(P<0.05)。结论:丹参酮IIA可抑制胃癌细胞的阿霉素耐药。
AIM:To investigate the inhibitory effect and the specific mechanism of tanshinone IIA on doxorubicin(DOX)-resistant gastric cancer cells.METHODS:The sensitivity of gastric cancer cells lines to DOX was determined by MTT assay.DOX-resistant gastric cancer cell lines were established by step selection with increasing concentrations of DOX.The cell cycle arrest,apoptosis and autophagy related-markers were analyzed by flow cytometry and Western blot.The expression of P-glycoprotein(P-gp),breast cancer resistance protein(BCRP)and multi-drug resistance-associated protein 1(MRP-1)was determined by RT-qPCR and Western blot.RESULTS:DOX-sensitive cell lines SNU-719 and SNU-601 as well as the cell lines relatively resistant to DOX including SNU-638,SNU-668,SNU-216 and SNU-620 were identified according to the IC 50 values of DOX for different cell lines.Two DOX-resistant cell lines SNU-719R and SNU-601R were also established.Tanshinone IIA inhibited the expression of MRP-1 in DOX-resistant cell lines.Compared with DOX treatment alone group,combined treatment of DOX and tanshinone IIA in cancer cells decreased the G 2/M phase cell number,increased the protein expression of p21,decreased the protein expressions of cyclin B1 and cyclin-dependent kinase 1(CDK1)in the SNU-719 R cells and SNU-620 cells.In addition,compared with DOX treatment alone group,combined treatment of DOX and tanshinone IIA in the cancer cells increased the protein expressions of p53,Bax and LC3B-II,decreased the protein expression of Bcl-2 and p62(P<0.05).CONCLUSION:Tanshinone IIA is an effective drug in the inhibition of DOX resistance in gastric cancer.
作者
刘源
倪渐凤芳
刘丽娜
李芳
尹先哲
LIU Yuan;NI Jian-feng;LIU Li-na;LI Fang-fang;YIN Xian-zhe(Department of Oncology,Nanyang Second General Hospital,Nanyang 473012,China;Department of Hematology,Nanyang Second General Hospital,Nanyang 473012,China;Department of Oncology,Nanyang First General Hospital,Nanyang 473000,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2019年第12期2208-2214,共7页
Chinese Journal of Pathophysiology
基金
河南省医学科技攻关计划项目(No.201602378)
