联合使用miR-34a及miR—let7对胰腺癌细胞生物学特性的影响  

Effect of the combination of miR-34a and miR-let7 on the biological properties of pancreatic cancer cells

作　　者：刘宇亭[1,3] 沈祥国[1] 苏长青[2] 孙斌[2] 李兆申[1] 徐灿[1] 

机构地区：[1]第二军医大学长海医院消化内科,上海200433 [2]第二军医大学东方肝胆外科医院分子肿瘤研究室 [3]东部战区空军机关医院

出　　处：《中华胰腺病杂志》2016年第2期87-92,共6页Chinese Journal of Pancreatology

基　　金：基金项目：国家自然科学基金面上项目（81372673）;上海市浦江人才计划项目（14PJD004）

摘　　要：目的探讨联合使用两种具有抑癌作用的microRNA（miRNA）同时转染人胰腺癌细胞对其生物学特性的影响。方法采用脂质体法将miR-34a及miR-let7单独或同时转染胰腺癌PANC1、SW1990细胞及正常胰腺腺泡AR42J细胞，以转染阴性对照miRNA（miR-NC）组及未转染组作为对照。应用qRT-PCR法检测各组细胞miR-34a及miR-let7的表达，MTT法检测细胞增殖，Transwell小室检测细胞迁移和侵袭能力，流式细胞仪检测细胞凋亡。结果MiR-34a转染组、miR-let7转染组、双转染组细胞的miR-34a、miR-let7表达水平均较miR-NC转染组及未转染组显著上调，差异有统计学意义（P值均〈0.05），表明miRNA成功转染了细胞。双转染组的PANC1、SW1990细胞增殖活性分别为（0.665±0.010）%、（0.638±0.030）%，较miR-NC转染组的（0.974±0.030）%、（0.971±0.050）%，miR-let7转染组的（0.888±0.050）%、（0.863±0.060）%显著被抑制，差异有统计学意义（P值均〈0.05），较miR-34a转染组的（0.795±0.060）%、（0.793±0.060）%下降，但差异无统计学意义。AR42J细胞的增殖活性无显著变化，差异无统计学意义。PANC1细胞miR-34a转染组、miR-let7转染组的穿膜细胞数分别为（103.70±3.28）、（100.70±1.76）个/200倍视野，较miR-NC转染组的（231.30±2.60）个/200倍视野及未转染组的（153.70±2.60）个/200倍视野显著减少，双转染组穿膜细胞数为（61.67±3.18）个/200倍视野，又较两个单转染组显著减少，差异均有统计学意义（P值均〈0.01）。细胞迁移实验与侵袭实验的结果一致。SW1990细胞侵袭及迁移能力的变化与PANC1一致。PANC1细胞的miR-34a转染组、miR-let7转染组、双转染组、miR-NC转染组、未转染组的细胞凋亡率分别为（16.66±1.27）%、（15.46±0.33）%、（23.35±1.80）%、（9.33±0.31）%、（8.83±0.36）%。两单转染组的细胞凋亡率较miR-NC转染组、未转染组显著增加；双转染组又Objective To investigate the influence on biological characteristics in human pancreatic cancer cells after beding transfected by two anti-carcinoma miRNAs at the same time. Methods Pancreatic cancer cells PANC1, SW1990 and normal pancreatic cells AR42J were transfected by miR-34a and（or） miR- let7 by liposome. Ceils transfected with negative control miRNA （miR-NC） and untransfected were as controls. The expression of miR-34a and miR-let7 were detected by real-time fluorescent quantitative RT-PCR. The cell proliferation was detected by MTT test and the migration and invasion were evaluated by transwell assay. The apoptosis rate was measured by flow cytometric analysis. Results After being transfected with miRNAs, the expression of miR-34a and miR-let7 in double transfection group （ miR-34a and miR-let7 were transfected at the same time）, miR-34a transfection group, miR-let7 transfection group was significantly up-regulated than those in miRNA-NC transfection group and untransfected group in PANC1 cells, SW1990 cells and AR42J cells, repectively. The difference which was statistically significant （P 〈0.05 ）indicating that cells were successfully transfected. The cell proliferation in double transfection group of PANC1 cells and SW1990 cells were （ 0. 665 ± 0.01, 0. 6375 ± 0.03 ） , which were significantly inhibited compared with （ 0. 974 ± 0.03, 0. 971 ± 0.05 ） in miR-NC group and （0.8875 ± 0.05, 0. 8625 ± 0.06） in miR-let7 group. The difference was statistically significant（P 〈0.05）. The cell proliferation activity in double transfection group was lower than those in miR-34a group （0. 795 ± 0.06, 0. 7925 ± 0.06）, but did not have statistically significant difference. There was no significant change in AR42J cells. Cell invasion assay showed that the number of PANC1 cells permeating substrate membrane in miR34a group （ 103.7 ± 3.28 ） and miR-let7 group （ 100.7 ± 1.76 ） were significantly fewer than ufiR-NC group （231.3 ±2.6） and untransfected group （

关 键 词：胰腺肿瘤 细胞系 肿瘤 MIR-34A miR-let7 转染 分子生物学 

分 类 号：R735.9[医药卫生—肿瘤]