MEK抑制剂在K-RAS基因突变的结直肠癌细胞中的耐药机制  

Effect of MEK inhibitor trametinib on drug resistance in K-RAS mutated colorectal cancer cells and its mechanism

作　　者：金剑英 朱延安 谢静静 金丹 JIN Jianying;ZHU Yan’an;XIE Jingjing(Department of Hematology and Oncology,Taizhou Hospital,Taizhou 317000,China)

机构地区：[1]台州医院血液肿瘤内科,317000 [2]台州医院急救中心,317000

出　　处：《浙江医学》2020年第2期109-113,共5页Zhejiang Medical Journal

基　　金：台州市市级科技资金补助项目（2016A33170）

摘　　要：目的探讨丝裂原激活蛋白激酶的激酶(MEK)抑制剂曲美替尼(Trametinib)在v-Ki-ras2 Kirsten大鼠肉瘤病毒癌基因同源物(K-RAS)基因突变的结直肠癌(CRC)细胞中的耐药机制。方法用Trametinib处理K-RAS基因突变型CRC细胞株LS174T、DLD-1、HCT116,Western blot法检测STAT3上游蛋白JAK2、JAK3和Src等是否激活。用Trametinib、JAK2/STAT3抑制剂鲁索替尼(Ruxolitinib)/LY5或者两者联用处理HCT116细胞,Western blot法检测p-STAT3Y705、STAT3、p-ERK1/2表达,磺酰罗丹明B蛋白染色实验观察细胞增殖情况。用Trametinib、LY5或Trametinib+LY5处理HCT116、DLD-1细胞,细胞迁移划痕实验观察细胞迁移情况,MTT实验检测细胞生存率。结果Trametinib能明显抑制ERK1/2的磷酸化,同时也能诱导p-STAT3Y705增加,诱导STAT3上游激酶p-JAK2高表达。Trametinib+Ruxolitinib/LY5联合用药,可同时阻断K-RAS基因突变的CRC细胞p-STAT3Y705、p-ERK1/2的激活,协同抑制肿瘤细胞的生长;可明显抑制肿瘤细胞克隆形成,减弱肿瘤细胞增殖能力;可抑制肿瘤细胞迁移能力,效果明显优于单药组(均P<0.05);可抑制细胞增殖,细胞生存率明显低于单药组(均P<0.05)。结论MEK和STAT3信号通路双重抑制的联合用药方案,是CRC治疗方案的新研究方向。Objective To investigate the effect of MEK inhibitor trimetinib on drug resistance in colorectal cancer(CRC)cells with v-Ki-ras2 Kirsten rat K-RAS gene mutation and its mechanism.Methods K-RAS mutant CRC cell lines LS174T,DLD-1,HCT116 were used to detect the activation of STAT3 upstream protein JAK2,JAK3 and Src by Western blot after treatment with Trametinib.The HCT-116 cells were treated with JAK2/STAT3 inhibitor ruxolitinib(LY5)and/or traminib,the cell proliferation and migration were examined.Results Trametinib significantly inhibited the phosphorylation of ERK1/2,induced the increase of p-STAT3Y705 and the high expression of p-JAK2 upstream kinase.Combined use of trametinib and ruxolitinib/LY5 blocked the activation of p STAT3Y705,p-ERK1/2 in CRC cells with K-RAS gene mutation,and inhibited the proliferation and clone formation of tumor cells,the effects were stronger than single drug use(P<0.05).Conclusion The study suggests that the combined application of MEK and STAT3 signal pathway inhibition may be a new research direction for the treatment of colorectal cancer.

关 键 词：丝裂原激活蛋白激酶的激酶抑制剂 曲美替尼 v-Ki-ras2 Kirsten大鼠肉瘤病毒癌基因同源物 结直肠癌 耐药 

分 类 号：R735.3[医药卫生—肿瘤]