MiR-214对人肺腺癌细胞PC9增殖及凋亡的研究

The Effect of MiR-214 on the Growth and Apoptosis of Human Lung Adenocarcinoma Cell Line PC9

[目的]探讨miR-214对肺腺癌细胞PC9增殖和凋亡的影响。[方法]培养人肺腺癌细胞株PC9(EGFR基因19外显子缺失,对EGFR-TKI吉非替尼敏感),将miR-214类似物(miR-214 mimic)及对照物(scramble)转染至PC9细胞中,在不同的时间点(24、48、72h)使用MTT检测细胞的增殖;转染细胞后,加入吉非替尼共培养48h后,使用流式细胞仪检测细胞凋亡;同时建立PC9荷瘤小鼠,观察荷瘤小鼠瘤体的生长曲线,及尾静脉注射miR-214抑制剂(antigomiR-214)后观察荷瘤小鼠瘤体的生长。[结果 ]在PC9细胞中,miR-214 mimic被转染入细胞后,48h及72h后细胞增殖率分别增加了27.9%和18.7%,相比转让了对照组的细胞差异均有统计学意义(P均<0.05)。吉非替尼可以诱导PC9细胞凋亡,但转染miR-214mimic后细胞凋亡率由86.5%下降为28.5%。在荷瘤小鼠中抑制miR-214后,小鼠瘤体明显缩小(P<0.05)。[结论]miR-214可促进肺腺癌细胞增殖及抗凋亡,可能可作为肺腺癌治疗的新靶点。

[Purpose] To investigate the effect of miR-214 on the growth and apoptosis of PC9 in vitro and in vivo. [Methods] PC9 cells（EGFR exon 19 del and sensitive to EGFR-TKI,such as gefitinib） were cultured and miR-214 mimic or scramble were transfected into PC9 cells;the growth was detected by MTT at different time points（24,48 and 72h）. Next,PC9 cells were induced apoptosis by gefitinib after 48 h of miR-214 or scramble transfection,and the apoptosis rate was analyzed by Annexin-FITC. The xenograft model of PC9 mice was established and miR-214 inhibitor were transfered into the mice,the xenograft tumor volume and growth curve were observed. [Results] Compared with the control group,the PC9 cells growth was increased 27.9% and18.7% when miR-214 mimic tranfected into PC9 cells after 48 or 72h（all P〈0.05）. Gefitinib could induce the apoptosis of PC9 cells,but with the transfection of miR-214 mimic,the apoptosis rate decreased from 86.5% to 28.5%. When miR-214 was inhibited by antagomir-214 in vivo,xenograft tumor volume decreased significantly（P〈0.05）. [Conclusion] Mi R-214 can promote the growth and reduce apoptosis of PC9 cells;it might be a potential therapeutic target lung adenocarcinoma in future.