摘要
目的观察微小RNA（miRNA，miR）-371—5p对人结直肠癌LoVo和SW620细胞增殖和凋亡的影响。方法构建miR-371-5p过表达慢病毒载体和稳定沉默miR-371—5p基因载体[miR-371-5p／短发卡RNA（shRNA）]，并转染LoVo和SW620细胞，反转录．聚合酶链反应（RT-PCR）测定miR-371-5p的表达，细胞计数试剂盒（CCK-8）、平板克隆形成实验、软琼脂集落形成实验、流式细胞仪检测LoVo和SW620细胞的增殖和凋亡。结果LoVo和SW620细胞转染miR-371—5p后miR-371—5p表达（14．25±0．25、10．01±0．19）高于阴性对照组和空白对照组（1．03±0．02、1．04±0．03；1．00±0．00、1．00±0．00；F=21．426、8．957，P=0．000、0．003），LoVo和SW620细胞转染miR-371—5p／shRNA后miR-371-5p表达（0．08±0．00、0．11±0．01）低于阴性对照组和空白对照组（0．97±0．02、1．04±0．03；1．00±0．00、1．00±0．00；F=15．231、8．537，P=0．000、0．005）。LoVo和SW620细胞转染后3d和7d，转染miR-371-5p的细胞增殖水平低于阴性对照组和空白对照组（F=5．932、19．324、7．256、35．846，P=0．003、0．000、0．000、0．000）；转染miR-371—5p／shRNA的细胞增殖高于阴性对照组和空白对照组（F=37．342、43．254、43．242、105．231，P=0．003、0．000、0．000、0．000）。转染miR-371—5p的LoVo和SW620细胞克隆个数低于阴性对照组和空白对照组（F=5．432、4．023，P=0．004、0．022）；转染miR-371-5p／shRNA的LoVo和SW620细胞克隆个数高于阴性对照组和空白对照组（F=3．976，4．732，P=0．015、0．011）。转染miR-371—5p的LoVo和SW620细胞集落个数低于阴性对照组和空白对照组（F=5．647、4．023，P=0．007、0．015）；转染miR-371-5p／shRNA的LoVo和SW620细胞集落个数高于阴性对照组和空白对照组（F=4．231、4．231，P=0．012、0．013）。转染miR-371—5p和miR．371—5p／shRNA的LoVo和SW620细胞凋亡率和阴性对照�
Objective To observe the effect of microRNA （miRNA, miR） -371 -5p on human colorectal cancer LoVo and SW620 cell proliferation and apoptosis. Methods Constructed miR -371 -5p overexpression lentivirus vector and stable gene silencing miR - 371 - 5p vector [ miR - 371 - 5p/short hairpin RNA （sbRNA） ] , and were transfected into LoVo and 5W620 cells. Reverse transcriptase -poly- merase chain reaction （RT- PCR） detected the expression of miR- 371 -5p, cell counting kit- 8 （ CCK - 8 ）, colony formation assay, soft agar colony formation assay, flow cytometry detected the prolifera- tion and apoptosis of LoVo and SW620 cells. Results The expression of miR - 371 - 5p in LoVo and SW620 cells transfected miR - 371 - 5p （ 14.25 ± 0. 25, 10.01 ± 0. 19） were higher than that of negative control group and blank control group （ 1.03 ± 0. 02, 1.04 ± 0. 03 ; 1.00 ± 0. 00, 1.00 ± 0. 00 ; F = 21. 426, 8. 957, P = 0. 000, 0. 003）, The expression of miR - 371 - 5p in LoVo and SW620 cells transfected miR-371 -5p/shRNA （0. 08 ±0. 00, 0. 11 ±0. 01 ） were lower than that of negative control group and blank control group cells （0.97 ± 0. 02, 1.04 ± 0. 03 ; 1.00 ± 0. 00, 1.00 ± 0. 00 ; F = 15. 231, 8. 537, P = 0. 000, 0. 005 ）. After LoVo and SW620 cells transfected 3 d and 7 d, the proliferation of miR - 371 - 5p transfected cell was lower than that of negative control group and blank control group （ F =5. 932, 19. 324, 7. 256, 35. 846, P = 0. 003, 0. 000, 0. 000, 0. 000） ; the proliferation of transfected miR -371 -5p/shRNA cell was higher than that of negative control group and blank control group （F = 37.342, 43.254, 43.242, 105.231, P=0.003, 0.000, 0.000, 0.000）. The cell clone number of miR- 371 -5p transfected LoVo and SW620 were lower than that of negative control group and blank control group cells （ F = 5. 432, 4. 023, P = 0. 004, 0. 022 ） ; the cell clone number of miR - 371 - 5p/ shRNA transfeeted LoVo and SW620 were higherthan that of negative control group and blank con
出处
《中华实验外科杂志》
CSCD
北大核心
2017年第6期982-984,共3页
Chinese Journal of Experimental Surgery
