miR-124在卵巢癌中的表达及其对卵巢癌细胞生物学行为的影响  

The expression of miR-124 in ovarian cancer tissues and its effect on biological functions of ovarian cancer cells

作　　者：舒锦[1] 袁犁[2] 刘兴明[3] 李少林[1] 周琦[1] 

机构地区：[1]重庆医科大学基础医学院放射医学教研室,重庆400016 [2]重庆市肿瘤研究所妇瘤科,重庆400030 [3]重庆市肿瘤研究所检验科,重庆400030

出　　处：《肿瘤》2014年第5期430-436,共7页Tumor

基　　金：重庆市科技攻关计划项目(编号:CSTC;2011AB5086)

摘　　要：目的:探讨微小RNA(microRNA,miRNA,miR)-124在卵巢癌组织中的表达及其对卵巢癌细胞生物学行为的影响。方法 :应用实时荧光定量PCR法检测35例卵巢癌组织及其邻近癌旁组织中miR-124的表达水平。将miR-124模拟物片段(has-miR-124 mimic)转染入卵巢癌SKOV3细胞中,应用CCK-8(cell counting kit-8)法和FCM法检测miR-124对卵巢癌SKOV3细胞增殖和凋亡的影响,Transwell小室法检测其对SKOV3细胞迁移和侵袭能力的影响,蛋白质印迹法检测SKOV3细胞中磷脂酰肌醇-3-激酶催化亚单位α(phosphatidylinositol-3 kinase catalytic subunit alpha,PIK3CA)、AKT和磷酸化AKT(phospho-AKT,p-AKT)蛋白表达水平的变化。结果:卵巢癌组织中miR-124的表达水平明显低于邻近的癌旁组织(P<0.05)。has-miR-124 mimic转染组SKOV3细胞的增殖、侵袭和迁移能力明显低于空白对照组(只转染lipofectamine)和阴性对照组[转染miR-124模拟物对照片段(has-miR-124 mimic control)](P<0.05),早期凋亡细胞所占比例明显增加(P<0.01),PIK3CA和p-AKT蛋白的表达水平明显下降(P<0.01),总AKT蛋白的表达水平无明显变化(P>0.05)。结论:卵巢癌组织中miR-124的表达水平下降。卵巢癌SKOV3细胞中miR-124过表达可明显抑制细胞的增殖、迁移和侵袭并诱导细胞凋亡,这一作用可能与抑制卵巢癌细胞中磷脂酰肌醇3-激酶(phosphatidylinositol-3 kinase,PI3K)/AKT信号通路的活性有关。Objective： To investigate the expression of miR-124 in ovarian cancer tissues and its effect on biological functions of ovarian cancer cells. Methods： The expression levels of miR-124 in ovarian cancer tissues and adjacent para-cancerous tissues from 35 cases were detected by real-time fluorescence quantitative-PCR. The SKOV3 cells were transfected with has-miR-124 mimic. Then the proliferation and apoptosis of the SKOV3 cells were detected by cell counting kit-8 （CCK-8） assay and flow cytometery, respectively, and the abilities of migration and invasion were examined by Transwell chamber assay. The expression levels of phosphatidylinositol-3 kinase, catalytic subunit alpha （PIK3CA）, AKT and phospho- AKT （p-AKT） proteins in SKOV3 cells were examined by Western blotting. Results： The expression level of miR-124 in ovarian cancer tissues was lower than that in adjacent para-cancerous tissues （P 〈 0.05）. The abilities of proliferation, migration and invasion of SKOV3 cells after transfection with has-miR-124 mimic were lower than those of the blank control cells （SKOV3 cells only transfected with lipofectamine） and the negative control cells （SKOV3 cells transfected with a has-miR-124 mimic control） （P 〈 0.05）, as well as the ratio of early apoptotic cells and expression levels of PIK3CA and p-AKT proteins were increased （all P 〈 0.01） whereas the expression level of total AKT protein had no significant change （P 〉 0.05）. Conclusion： The expression level of miR-124 is significantly decreased in ovarian cancer tissues. Over-expression of miR-124 in SKOV3 cells can suppress the abilities of proliferation, migration and invasion, and induce apoptosis. This effect may be related to the suppression of PI3K/AKT signaling pathway.

关 键 词：卵巢肿瘤 微RNAS 细胞增殖 细胞凋亡 miR-124 细胞 SKOV3 

分 类 号：R737.31[医药卫生—肿瘤]