靶向于人端粒酶逆转录酶的miR-138增强结肠癌细胞SW480对5-氟尿嘧啶的敏感性

MiR-138 targeting human telomerase revegse transcriptase enhanced the sensitization of colon canc- er cell SW480 to 5-fluorouracil

目的观察miR-138对结肠癌细胞SW480人端粒酶逆转录酶（hTERT）表达的影响，探讨miR-138模拟体mimic沉默hTERT在SW480对5-氟尿嘧啶（5-Fu）化疗药物敏感性中的作用。方法流式细胞仪检测50nmol／L和10nmol／LmiR-138模拟体（mimic）转染SW480后hTERT的表达；5μl异硫氰酸荧光素（FITC）标记的细胞核增殖抗原（Ki-67）抗体染色转染和未转染50nmol／Lmimic的SW480，流式细胞仪检测1mmol／L5-Fu处理和未处理的细胞增殖；碘化丙锭（PI）单染和膜联蛋白V（AnnexinV）-PI双染转染和未转染50nmol／Lmimic的SW480，流式细胞仪检测1mmol／L5-Fu处理和未处理的细胞周期和凋亡状态。结果miR-138mimic在50nmol／L和10nmol／L时，hTERT表达率分别为（30．25±1．34）％和（60．03±2．78）％，而阴性对照组为（87．50±1．63）％；转染miR-138mimic的SW480细胞组与非转染组Ki-67阳性细胞分别为（58．18±2．90）％和（83．45±2．41）％；sub．Go期分别为（27．33±1．70）％和（1．05±0．19）％；凋亡率达（29．50±2．27）％和（5．30±1．74）％；mimic转染组、5-Fu处理组和mimic转染与5-Fu联合组的细胞增殖率分别为（61．00±1．73）％、（57．00±2．03）％和（23．00±1．24）％，sub-G0期细胞分别为（28．12±1．47）％、（34．87±2．01）％和（70．94±3．04）％；凋亡细胞比例分别为（32．15±2．76）％、（40．07±2．58）％和（80．84±3．06）％，差异均有统计学意义（P〈0．05）。结论MiR-138mimic可以抑制SW480中hTERT表达并增强SW480对5-Fu的敏感性。

Objective To observe the effect of miR-138 on the expression of human telomerase re- verse transcriptase （hTERT） in human colon cancer cell SW480 and to evaluate influence of miR-138 mimic on the sensitization of SW480 to 5-fluorouracil （5-Fu）. Methods Flow cytometry was used to detect the expression of hTERT in SW48 cells transfected with 50 and 10 nmol/L miR-138 mimic. Both SW480 cells transfected and un-transfected with 50 nmol./L miR-138 mimic were stained by FITC-labeled Ki-67 antibody and assayed by using flow cytometry. The apoptosis of both SW480 cells transfected and un- transfected with 50 nmol/L miR-138 mimic was tested by using PI single dye method and Annexin V-P! ap- optosis assay. Results miR-138 mimic at 50 and 10 nmol/L down-regulated the expression of hTERT at （ 30. 25 ± 1.34） % and （ 60.03 ± 2.78 ） % respectively, and the expression rate of hTERT in the control group was （87. 50 ± 1.63 ）%. In SW480 cells transfected or un-transfected with 50 nmol/L of miR-138 mimic, the Ki-67 positive rate was （58.18 ±2. 90）% was （83.45 ±2.41 ）%, sub-G0 rate was （27. 33 ± 1.70）% and （1.05 ±0.19）%, and apoptosis rate was （29.50 ±2.27）% and （5.30 ± 1.74）%, respectively. In miR-138 mimic-transfected group, 5-Fu-treated group, and combined treatment group, proliferation rate was （61.00 ± 1.73）% , （57.00 ±2.03）% and （23.00 ± 1.24）% , sub-G0 rate was （28. 12±1.47）%, （34.87 ±2.01）% and （70.94 ±3.04）%, and apoptosis rate was （32.15± 2. 76）% , （40. 07 ±2. 58）% and （80. 84±3.06）% in SW480 cells, respectively. All differences were statistically significant （ P 〈 0. 05 ）. Conclusion miR-138 mimic inhibited the expression of hTERT in SW480 cells, and enhanced sensitization of SW480 cells to 5-Fu.