miR-223对结肠癌SW480细胞生物学行为的影响

目的 观察miR-223对结肠癌SW480细胞增殖、细胞周期及凋亡的影响.方法 PLL3.7-miR-223和PLL3.7-miR-EV质粒转染SW480细胞,Real-time PCR检测转染24h后miR-223的表达变化,CCK-8试剂盒检测转染12、24、48 h后细胞的增殖能力,流式细胞仪分析转染48 h后细胞周期和凋亡情况,Western blot检测IGF-1R、AKT蛋白表达情况.结果 转染24 h后,Real-time PCR检测结果显示PLL3.7-miR-223转染组miR-223的表达量（6.55±2.04）较PLL3.7-miR-EV（以其miR-223的表达量为1）明显上调（P＜0.01）.与对照EV组相比,SW480细胞在转染miR-223后生长明显受到抑制,细胞周期在G1期发生阻滞（61.61±4.02）％vs.（35.99±1.62）％],凋亡增加明显（67.43±4.75）％vs.（44.23±3.11）％],均P＜0.01,并且能够降低生长相关蛋白IGF-1R及细胞凋亡相关蛋白AKT的表达.结论 miR-223可以通过调节IGF-1R和AKT的表达影响结肠癌细胞SW480的增殖、细胞周期和凋亡.

Effect of miR-223 in the biological behavior of colon cancer cell line SW480

Objective To investigate the role of miR-223 on the proliferation,cell cycle and apoptosis of colon cancer cell line SW480.Methods PLL3.7-miR-223 and PLL3.7-miR-EV plasmid were transiently transfected into SW480 cells.The expression of miR-223 was examined using Real-time PCR.The proliferation of cells were detected by using Cell Counting Kit-8 at respective time of 12,24,48 hours after tansfection.The cell cycle and apoptosis of transfected cells were detected by flow cytometry assay.The levels of IGF-1R and AKT protein expression were determined by Western blot.Results 24 hours after transfection,the expression of miR-223 in PLL3.7-miR-223 transfected group （6.55 ± 2.04）was higher than that of PLL3.7-miR-EV group,which was defined as 1 （P 〈0.01）.Compared with the control group,the miR-223 group showed a significant inhibition of proliferation,G1 arrest （61.61 ± 4.02） % vs.（35.99 ± 1.62） %] and an increased rate of apoptosis （67.43 ± 4.75） % vs.（44.23 ± 3.11） %],all P 〈 0.01.The expression of IGF-1R and AKT of miR-223 group were lower than that of EV group.Conclusions In this study,we found that miR-223 could inhibit the proliferation,promote G1 arrest and apoptosis of colorectal cell line SW480 through down-regulate IGF-1R and AKT.