肝再生磷酸酶-3-信号传导和转录激活子3-微小RNA21信号通路促进结肠癌细胞增殖侵袭的研究

Phosphatase of regenerating liver-3-signal transducer and activators of transcription 3-miR-21 sig- nal pathway promotes proliferation and invasion of colon cancer cells

目的观察肝再生磷酸酶-3-信号传导和转录激活子3-微小RNA21（PRL-3-STAT3-miR-21）信号通路对结肠癌细胞增殖侵袭的作用。方法构建稳定转染PRL-3基因和空白对照质粒的结肠癌细胞株LoVo、PRL-3和LoVo．VC，用荧光实时定量聚合酶链反应（RT—qPCR）检测稳转细胞株中miR-21的表达并在瞬时转染PRL-3的SW480及CaC02细胞中进行验证。用Westernblot法对PRL-3调控STAT3的表达进行检测，在LoVo-PRL-3细胞中对STAT3进行RNA干扰，检测miR-21的表达，在稳转细胞株中转染miR-21或对其进行敲除，用细胞计数试剂盒（CCK-8）、Transwell实验对细胞的增殖侵袭能力的变化进行研究。结果LoVo—PRL-3细胞在72、96h的增殖能力以及在Tran-swell实验24h后的侵袭能力要强于对照组LoVo—VC细胞（P〈0．01）。在结肠癌细胞株LoVo—PRL-3中pSTAT3、miR-21的表达明显上调，干扰STAT3可以抑制miR-21的表达（P〈0．05）。在LoVo—VC细胞中过表达miR-21促进了细胞的增殖侵袭（P〈0．01），而在LoVo-PRL-3细胞中敲除miR-21抑制了细胞的增殖侵袭（P〈0．05）。结论PRL-3-STAT3-miR-21信号通路在结肠癌细胞增殖侵袭中起促进作用。

Objective To explore the effect of phosphatase of regenerating liver-3-signal transducer and activators of transcription 3-microRNA-21 （ PRL-3-STAT3-miR-21 ） signal pathway on proliferation and invasion of colon cancer cellsMethods We stablely transfeeted PRL-3 expressing plasmid and empty plasmid into LoVo colon cancer cells and established two cell lines： LoVo-PRL-3 and LoVo-VC. We used quantitative real-time reverse transcription-polymerase chain reaction （qRT-PCR） to detect the expression of miR-21 in LoVo cells. We also performed transient transfection of PRL-3 into SW480 and CACO2 cells to validate the expression of miR-21. Western blotting was used to detect the effect of PRL-3 on the expres- sion of STAT3. RNA interference was used to knocked down STAT3 in LoVo-PRL-3 cells, after that miR- 21 expression was detected. Transient transfection of miR-21mimic into LoVo-VC cells or transient trans- fection of miR-21 inhibitor into LoVo-PRL-3 cells were performed to evaluate the proliferation and invasive ability of these ceils by CCK8 proliferating assay and Transwell chamber assay. Results PRL-3 promoted proliferation of LoVo-PRL-3 cells at 72 h, 96 h and invasion of LoVo-PRL-3 cells after 24 h compared with LoVo-VC cells （P 〈0. 01）. In LoVo-PRL-3 cells, pSTAT3 and miR-21 were up-regulated. Knock-down of STAT3 mRNA decreased the expressign of miR-21 （P 〈 0. 05 ）. Over-expression of miR-21 promoted proliferation and invasion of LoVo-VC cells （P 〈 0. 01 ）, while knocking down of miR-21 inhibited prolifer- ation and invasion of LoVo-PRL-3 ceils （P 〈 0. 05）. Conclusion PRL-3-STAT3-miR-21 signal pathway promotes proliferation and invasion of colon cancer ceils.