摘要
目的:探讨miR-29c调控TNRC18胃癌组织和细胞阿帕替尼耐药性的机制。方法:收集2015年2月至2017年10月武汉市中心医院具有完整资料的39例胃癌和癌旁组织标本(其中21例为阿帕替尼耐药患者、18例为不耐药患者),采用qRT-PCR检测miR-29c在胃癌组织和细胞系中的表达水平。采用CCK-8、Transwell和Annexin V-FITC/PI双染流式术检测miR-29c过表达/敲降对MGC-803/AP耐药细胞增殖、侵袭和凋亡影响,Western blotting检测miR-29c调控TNRC18表达,双荧光素酶报告基因验证miR-29c与TNRC18的靶向作用关系。结果:miR-29c在3种胃癌细胞系和阿帕替尼耐药癌患者组织中均低表达。双荧光素酶报告基因证实miR-29c靶向作用TNRC18并下调其表达水平。miR-29c通过靶向下调TNRC18抑制阿帕替药耐药的MGC-803/AP细胞的增殖、侵袭并促进细胞凋亡(均P<0.05或P<0.01),进而降低胃癌细胞MGC-803/AP对阿帕替尼的耐药性。体内实验同样证实,miR-29c通过靶向抑制TNRC18降低胃癌对阿帕替尼的耐药性。结论:miR-29c/TNRC18分子轴在胃癌组织和细胞MGC-803/AP对阿帕替尼耐药中发挥着一定作用,过表达miR-29c可逆转MGC-803/AP细胞对阿帕替尼耐药。
Objective:To investigate the mechanism of miR-29c modulating apatinib resistance of gastric cancer tissues and cells MGC-803 via regulating TNRC18.Methods:A total of 39 gastric cancer patients with complete clinical data,who were treated in the Central Hospital of Wnhan from Feb.2015 to Oct.2017,were collected for this study.The expression of miR-29c was detected by qRT- PCR in gastric cancer tissues and cell lines.The effect of miR-29c over-expression/knockdown on the proliferation,invasion and apop- tosis of MGC-803/AP cells in vitro was measured by CCK-8assay,Transwell and Annexin V-FITC/PI double staining flow eytometry assay.Western blotting was used to detect the regulation of miR-29c on TNRC18.Moreover,the relationship between miR-29c and TN- RC18was examined by dual luciferase reporter gene assay.Results:qRT-PCR revealed that miR-29c was low expressed in gastric can- cer cell lines and gastric cancer tissues from patients resistant to apatinib.Moreover,dual luciferase reporter gene assay confirmed that miR-29c directly binds to the 3'UTR of TNRC18mRNA to suppress its expression in MGC-803/AP cells.Furthermore,miR-29c inhibited the apatinib resistance in gastric cancer MGC-803/AP cells via inhibiting cell proliferation,invasion and promoting cell apoptosis by targeted down-regulating TNRC18.Additionally,in vivo experiment also confirmed that miR-29c modulated apatinibTresistance in gastric cancer cells by targeted inhibiting TNRC18.Conclusion:miR-29c/TNRC18axis plays a certain role in the resistance of gastric cancer tissues and MGC-803/AP cells to apatinib,and over-expression of miR-29c may reverse the resistance of MGC-803/AP cells to apatinib.
作者
陈志刚
卢宏达
唐求
CHEN Zhigang;LU Hongda;TANG Qiu(Department of Oncology,the Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,Hubei,China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2018年第11期1140-1147,共8页
Chinese Journal of Cancer Biotherapy
