hsa-miR-206过表达调节MAPK通路抑制卵巢癌细胞CAOV3生长及运动  

Overexpression of hsa-miR-206 inhibits the growth and movement of ovarian cancer cell CAOV3 by regulating the MAPK pathway

作　　者：徐瑶 邓晓杨[2] 张勇 吴雯[1] 肖琳 张小虎 Xu Yao;Deng Xiaoyang;Zhang Yong(Dept of Gynecology,Affiliated Hospital of Southwest Medical University/The First Affiliated Hospital of Chengdu Medical College,Luzhou 646000;Dept of Gynecology,The First Affiliated Hospital of Chengdu Medical College,Chengdu 610500;Dept of Obstetrics and Gynecology,Mianyang Central Hospital,Mianyang 621000)

机构地区：[1]西南医科大学附属医院/成都医学院第一附属医院妇科,泸州646000 [2]成都医学院第一附属医院妇科,成都610500 [3]绵阳市中心医院妇产科,绵阳621000

出　　处：《安徽医科大学学报》2020年第6期848-854,共7页Acta Universitatis Medicinalis Anhui

基　　金：四川省教育厅自然科学重点项目(编号:17ZA0128)。

摘　　要：目的探讨微小型RNA miR-206对卵巢癌细胞CAOV3生长及运动特性影响及作用机制。方法生物信息预测miR-206靶向基因,荧光素酶实验验证其靶向关系。细胞分为Control、miR-206 mimic、pc-MAPK1及miR-206 mimic+pc-MAPK1组,qRT-PCR检测miR-206表达,Brdu染色检测细胞生长,Transwell检测细胞侵袭,Western blot法检测促分裂原活化蛋白激酶1 (MAPK1)、上皮型钙黏附蛋白(E-cadherin)、波形蛋白(vimentin)、cAMP反应元件结合蛋白(CREB)、磷酸化CREB(P-CREB)、Ets样转录因子-1(Elk-1)、P-Elk-1、c-Myc、P-c-Myc及血管内皮生长因子(VEGF)表达。建立经或未经miR-206转染的CAOV3细胞裸鼠皮下移植瘤,测量移植瘤体积,qRT-PCR检测miR-206及MAPK1表达水平,Western blot法检测Elk-1、P-Elk-1、原癌基因(c-Myc)及P-c-Myc表达,免疫组化检测Ki67及vimentin表达。结果 miR-206靶向抑制MAPK1;与Control组相比,miR-206 mimic组BrdU阳性细胞百分比、侵袭细胞数降低(t=4.27,P=0.002;t=3.43,P=0.008),E-cadherin表达提高(t=4.13,P=0.003),vimentin表达、P-CREB/CREB、P-Elk-1/Elk-1及P-c-Myc/c-Myc比值下调(t=3.51,P=0.007;t=3.27,P=0.010;t=3.26,P=0.010;t=3.28,P=0.010)。miR-206 mimic减小裸鼠皮下CAOV3细胞移植瘤体积(t=6.51,P<0.001),提高移植瘤组织中miR-206表达(t=4.65,P=0.001),降低MAPK1 mRNA水平、VEGF表达、P-Elk-1/Elk-1及P-c-Myc/c-Myc比值(t=3.86,P=0.004;t=3.37,P=0.009;t=3.34,P=0.009;t=3.40,P=0.008),减少Ki67及Vimentin阳性细胞百分比(t=4.45,P=0.002;t=4.22,P=0.002)。结论过表达miR-206可通过阻碍MAPK通路活化抑制卵巢癌CAOV3细胞生长及运动。Objective To investigate the effect and mechanism of microRNA miR-206 on the growth and motility of ovarian cancer cell CAOV3.Methods The targeted gene of miR-206 was predicted by bioinformatics and its targeted-relationship were confirmed by the luciferase reporter assay.Cells were divided into Control,miR-206 mimic,pc-MAPK1 and miR-206 mimic+pc-MAPK1 group,the expression of miR-206 was measured by qRT-PCR,cell growth was measured by Brdu staining,cell invasion was measured by Transwell,the expression of Mitogen-activated protein kinase 1(MAPK1),epithelial cadherin(E-cadherin),vimentin(vimentin),c AMP response element binding protein(CREB),phosphorylated CREB(P-CREB),Ets-like Transcription factor-1(Elk-1),P-Elk-1,proto-oncogene(c-Myc),P-c-Myc and vascular endothelial growth factor(VEGF)were measured by Western blot.Subcutaneous xenograft tumors of nude mice with miR-206 transfected or non-transfected CAOV3 cells were established.The volume of the transplanted tumors was measured.The expression levels of miR-206 and MAPK1 were detected by qRT-PCR.The expression levels of Elk-1,P-Elk-1,c-Myc and Pc-Myc expression was detected by Western blot.Ki67 and vimentin expression were detected by immunohistochemistry.Results miR-206 targeted the inhibition of MAPK1 expression.Compared with the control group,the percentage of Brd U positive cells and the number of invasive cells in the miR-206 mimic group reduced(t=4.27,P=0.002;t=3.43,P=0.008),the expression of E-cadherin increased(t=4.13,P=0.003),P-CREB/CREB,P-Elk-1/Elk-1,Pc-Myc/c-Myc ratios and the expression of vimentin were down-regulated(t=3.27,P=0.010;t=3.26,P=0.010;t=3.28,P=0.010,t=3.51,P=0.007).miR-206 mimic reduced the volume of subcutaneously transplanted tumors of CAOV3 cells in nude mice(t=6.51,P<0.001),increased the expression of miR-206 in the tissues of transplanted tumors(t=4.65,P=0.001),reduced MAPK1 mRNA levels,VEGF expression,P-Elk-1/Elk-1 and Pc-Myc/c-Myc ratio(t=3.86,P=0.004;t=3.37,P=0.009;t=3.34,P=0.009;t=3.40,P=0.008),and reduced the percentage of Ki67

关 键 词：miR-206 卵巢癌 MAPK通路 生长 运动 

分 类 号：R737.31[医药卫生—肿瘤]