摘要
目的:探讨FOXO1和Smad4协同抑制前列腺癌进展和转移的作用机制。方法:体外培养前列腺癌PC3细胞,采用脂质体转染方法将pEGFP-N1,pEGFP-FOXO1,pDsRed2-Smad4分别转染入PC3细胞,并依次分为pEGFP-N1组、pEGFP-FOXO1组、pDsRed2-Smad4组和pEGFP-FOXO1+pDsRed2-Smad4组4组,采用蛋白质印迹法和qPCR检测FOXO1和Smad4的蛋白和mRNA表达,CCK-8法检测细胞增殖活性,细胞划痕法检测细胞的迁移能力。结果:质粒成功转染,与pEGFP-N1组比,pEGFP-FOXO1组、pDsRed2-Smad4组和pEGFP-FOXO1+pDsRed2-Smad4组PC3细胞增殖能力、迁移能力显著降低,差异有统计学意义(P<0.05)。PC3细胞中过表达FOXO1可以上调Smad4的mRNA和蛋白表达水平,过表达Smad4可以明显增强FOXO1对PC3细胞增殖和迁移的抑制作用。结论:FOXO1与Smad4在前列腺癌中具有协同抑制前列腺癌增殖及转移的作用。
Objective:To investigate the mechanism of synergistic inhibition of FOXO1 and Smad4 on prostate cancer progression and metastasis.Methods:Prostate PC3 cells were cultured in vitro,and pEGFP-N1,pEGFPFOXO1 and pDsRed2-Smad4 were transfected into PC3 cells by lipofection.They were divided into 4 groups:pEGFP-N1 group,pEGFP-FOXO1 group,pDsRed2-Smad4 group and pEGFP-FOXO1+pDsRed2-Smad4 group.The protein and mRNA expressions of FOXO1 and Smad4 were detected by Western blotting and qPCR in the group.Cell proliferation activity was detected by CCK-8 method,and cell migration ability was detected by cell scratch method.Results:The plasmid was successfully transfected.Compared with the pEGFP-N1 group,the cell proliferation and migration ability of the pEGFP-FOXO1 group,pDsRed2-Smad4 group and pEGFPFOXO1+pDsRed2-Smad4 group were significantly decreased(P<0.05).Overexpression of FOXO1 in prostate cancer cell PC3 up-regulated the mRNA and protein expression levels of Smad4.Overexpression of Smad4 significantly enhanced the inhibitory effect of FOXO1 on proliferation and migration of PC3 cells.Conclusion:FOXO1 and Smad4 have synergistic effects in prostate cancer,synergistically inhibit prostate cancer proliferation and metastasis.
作者
瞿根义
张玉龙
徐勇
阳光
聂海波
QU Genyi;ZHANG Yulong;XU Yong;YANG Guang;NIE Haibo(Department of Urology,Affiliated Zhuzhou Hospital,Xiangya Medical College of Central South University,Zhuzhou Hunan 412007,China)
出处
《临床与病理杂志》
2020年第7期1625-1631,共7页
Journal of Clinical and Pathological Research
基金
湖南省科技创新计划项目(2017SK51003)
株洲市科技指导性计划项目(20180004)。
关键词
FOXO1
SMAD4
前列腺癌
协同
转移
forkhead transcription factor O1
Smad4
prostate cancer
synergy
metastasis
