摘要
目的观察miR-106b在非小细胞肺癌细胞A549中的表达变化,并探讨其对细胞迁移、侵袭的影响及可能机制。方法采用荧光定量PCR法检测非小细胞肺癌细胞A549及正常肺表皮细胞系HPL-1中miR-106b相对表达量。将非小细胞肺癌细胞A549分为miR-106b模拟物组、miR-106b抑制物组和阴性对照组,采用Lipofectamine2000转染法分别转染miR-106b mimics、miR-106b inhibit及scramble。培养24 h,采用荧光定量PCR法检测miR-106b相对表达量,进行细胞划痕试验、Transwell细胞侵袭试验检测细胞迁移和侵袭能力(分别以划痕愈合率、穿膜细胞数表示),采用荧光定量PCR法和Western blotting法检测墨角藻糖基转移酶6(FUT6)mRNA及蛋白表达。结果非小细胞肺癌细胞A549及正常肺表皮细胞HPL-1中miR-106b相对表达量分别为5.37±0.42、1.00,二者比较P<0.01。miR-106b模拟物组、miR-106b抑制物组及阴性对照组miR-106b相对表达量分别为8.94±0.73、0.21±0.03、1.00,组间两两比较P均<0.05。miR-106b模拟物组、阴性对照组划痕愈合率、穿膜细胞数均高于miR-106b抑制物组,FUT6 mRNA及蛋白相对表达量均低于miR-106b抑制物组,但miR-106b模拟物组变化更明显(P均<0.05)。结论非小细胞肺癌细胞A549中miR-106b表达升高;过表达miR-106b可能通过下调FUT6表达而促进非小细胞肺癌细胞的迁移与侵袭。
Objective To observe the expression changes of miR-106b in non-small-cell lung cancer （NSCLC） cell line A549, and to investigate its effects on cell migration and invasion, as well as the regulatory mechanism. Methods The expression level of miR-106b was measured in A549 and HPL-1 cells by the fluorescent quantitative PCR. A549 cells were divided into three groups. The miR-106b mimics group was transfeeted with miR-106b mimics, the miR- 106b inhibition group with miR-106b inhibit, and the control group with scramble. The relative expression level of miR- 106b was measured by real-time fluorescent quantitative PCR （qRT-PCR） at 24 h after culture. The migration and invasion abilities in the three groups were measured by Scratch test and Transwell invasion assay. The expression levels of FUT6 mRNA and protein were measured in these three groups by fluorescent quantitative PCR and Western blotting, respectively. Results The expression level of miR-106b in the A549 cells was 5.37 ± 0.42, which was higher than that of HPL-1 cells （1.0） , P 〈0.01. The expression level of miR-106b was 8.94 ±0.73 in the miR-106b mimics group, 0.21 ±0.03 in the miR-lO6b inhibit group and 1.0 in the control group, respectively, and the difference was significant, all P 〈 0.05. The wound healing rate and the number of invasive cells in the miR-106b mimics group and control group were significantly higher than those of the miR-106 inhibition group, while the expression levels of FUT6 mRNA and protein were significantly lower than those in the miR-106b inhibit group, and the change in the miR-106b mimics group was more significance （ all P 〈0.05）. Conclusion The miR-106b is up-regulated in the NSCLC A549 cells and the over-expression of miR-106b promotes the migration and invasion of NSCLC by down-regulating FUT6 expression.
出处
《山东医药》
CAS
北大核心
2017年第44期22-25,共4页
Shandong Medical Journal
基金
武汉市应用基础研究计划项目
