沉默Lin28b基因表达促进人结肠癌细胞奥沙利铂化疗敏感性  

Silencing Lin28b promotes the chemosensitivity of human colon cancer cells to oxaliplatin

作　　者：庞明辉[1,2] 王康[2] 伍刚[2] 梁耀泽[1] 胡阳[2] 何飞[3] 侯能易[2] 李平[2] 李国新[1] 

机构地区：[1]南方医科大学南方医院普外科,广州510515 [2]四川省人民医院普外科 [3]四川省人民医院人类疾病基因研究四川省重点实验室

出　　处：《中华实验外科杂志》2012年第7期1324-1327,共4页Chinese Journal of Experimental Surgery

基　　金：四川省卫生厅科研资助项目（110167）

摘　　要：目的探讨人结肠癌细胞中Lin28b基因表达及其与奥沙利铂化疗敏感性之间的关系。方法构建干扰Lin28b的shRNA质粒（sh-Lin28b），并转染至结肠癌细胞（Cac02、SW480和HCT116细胞）；逆转录-聚合酶链反应（RT—PCR）和Westernblot法分别检测结肠癌细胞中Lin28b在mRNA和蛋白水平的表达；用细胞计数试剂盒（CCK-8）检测sh—Lin28b和奥沙利铂化疗协同作用，Transwell实验检测处理后的结肠癌细胞迁移能力。结果RT—PCR和Western blot结果证实Cac02、HCT116及SW480结肠癌细胞中均有Lin28b基因表达，其中在SW480细胞中的蛋白表达量比在HCT116中高1．83倍。SW480和HCT116细胞中Lin28b的mRNA表达在50nmol／L的sh—Lin28b作用下分别被下调36．4％和90．2％，在100nmol／L的sh—Lin28b的条件下分别被下调61．8％和89．5％。CCK-8结果显示奥沙利铂处理48h后，HCT116细胞的IC50值为（6．09±1．42）mg／L，SW480细胞升高34．3％，至（8．18±3．64）mg／L。sh—Lin28b联合奥沙利铂处理组比较于对照转染组（NC）抑制SW480和HCT116细胞生存率分别为（55．10±0．78）％和（50．30±0．69）％。Transwell实验结果显示SW480细胞的迁移能力在sh—Lin28b的作用下降低至（48．60±0．92）％。结论沉默表达于结肠癌细胞的Lin28b，可显著抑制结肠癌细胞的迁移能力，并促进奥沙利铂的化疗敏感性。Objective To investigate the effects of Lin28b expression on the chemosensitivity of colon cancer cells to oxaliplatin. Methods Basic Lin28b mRNA and protein expression level was examined in colon cancer cells by reverse transcription-polymerase chain reaction （RT-PCR） and Western blotting, respectively. The half maximal inhibitory concentration of colon cancer cells （ICso） tranfected with sh- Lin28b and treated by oxaliplatin was calculated using cell counting Kit-8 （CCK-8） technique. The migra- tion of theses cells was determined with Transwell method to explore the synergistic effect of sh-Lin28b ex- pression and oxaliplatin. Results There was Lin28b expression in Caco2, HCT116 and SW480 colon cancer cell lines detected by RT-PCR and Western blotting. The protein expression level of Lin28b in SW480 cells was 1.83 times higher than that in HCTll6 cells. The treatment of 50 nmol/L sh-Lin28b re- sulted in 36. 4% and 90. 2% reduction of mRNA expression level of Lin28b in SW480 cells and HCT116 cells, respectively, and that of 100 nmol/L sh-Lin28b led to 61.8% and 89. 5% reduction, respectively. CCK-8 results showed that the ICS0 （6. 09 ± 1.42） mg/L of HCTll6 cells treated with oxaliplatin by 48 h was increased to 34. 3% （8.18 ±3.64） mg/L in SW480 cells. The half maximal inhibitory concentration of transfected SW480 and HCTll6 cells was decreased to （55.10 ±0. 78）% and （50. 30 ±0. 69）% by treatment with oxaliplatin as compared with that in normal control group. The Transwell results showed that the migration of transfected SW480 cells was reduced to （48.60 ±0.92 ） %. Conclusion Silencing the ex- pression of Lin28b in colon cancer cells could greatly inhibit their migration and promote their chemosensi- tivity to oxaliplatin.

关 键 词：Lin28b 结肠癌 奥沙利铂 化疗 

分 类 号：R735.35[医药卫生—肿瘤]