CAJ | 학술논문

目的探讨肝再生磷酸酶-3(PRL-3)和微小RNA17-92 (miR-17-92)家族成员在结肠癌中异常表达的意义.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MicroRNA芯片筛查表达异常的促癌microRNA,从中选取miR-17-92家族成员miR-17、miR-19a行荧光实时定量聚合酶链反应(qRT-PCR)进行验证.在LoVo-PRL-3细胞中对STAT3信号传导与转录激活因子-3(STAT3)进行RNA干扰,检测miR-17、miR-19a的表达,在稳转细胞株中转染miR-17、miR-19a或对其进行敲除,用细胞计数试剂盒(CCK-8)、Tanswell试验对细胞增殖侵袭能力的变化进行研究.在13例患者结肠癌原发灶、转移灶及癌旁正常组织中行免疫组织化学及qRT-PCR检测PRL-3、pSTAT3和miR-17、miR-19a的表达.结果在结肠癌细胞株LoVo-PRL-3中miR-17、miR-19a的表达明显上调(P＜0.05),干扰STAT3可以抑制miR-17、miR-19a的表达.在LoVo-VC细胞中过表达miR-17、miR-19a促进了细胞的增殖(P＜0.05及P＜0.01)和侵袭(P＜0.01),而在LoVo-PRL-3细胞中敲除miR-17、miR-19a抑制了细胞的增殖侵袭(P＜0.05).在结肠癌组织中PRL-3、pSTAT3和miR-17、miR-19a的表达呈正相关.结论 PRL-3通过上调miR-17、miR-19a的表达在结肠癌细胞增殖侵袭中起到促进作用.
목적 탐토간재생린산매-3(PRL-3)화미소RNA17-92 (miR-17-92)가족성원재결장암중이상표체적의의.방법 구건은정전염PRL-3기인화공백대조질립적결장암세포주LoVo-PRL-3화LoVo-VC,용MicroRNA심편사사표체이상적촉암microRNA,종중선취miR-17-92가족성원miR-17、miR-19a행형광실시정량취합매련반응(qRT-PCR)진행험증.재LoVo-PRL-3세포중대STAT3신호전도여전록격활인자-3(STAT3)진행RNA간우,검측miR-17、miR-19a적표체,재은전세포주중전염miR-17、miR-19a혹대기진행고제,용세포계수시제합(CCK-8)、Tanswell시험대세포증식침습능력적변화진행연구.재13례환자결장암원발조、전이조급암방정상조직중행면역조직화학급qRT-PCR검측PRL-3、pSTAT3화miR-17、miR-19a적표체.결과 재결장암세포주LoVo-PRL-3중miR-17、miR-19a적표체명현상조(P＜0.05),간우STAT3가이억제miR-17、miR-19a적표체.재LoVo-VC세포중과표체miR-17、miR-19a촉진료세포적증식(P＜0.05급P＜0.01)화침습(P＜0.01),이재LoVo-PRL-3세포중고제miR-17、miR-19a억제료세포적증식침습(P＜0.05).재결장암조직중PRL-3、pSTAT3화miR-17、miR-19a적표체정정상관.결론 PRL-3통과상조miR-17、miR-19a적표체재결장암세포증식침습중기도촉진작용.
Objective To explore the significance of the aberrant expression phosphatase of regenerating liver-3 (PRL-3) and microRNA (miR)-17-92 family in colon cancer cells.Methods We stablely transfected PRL-3 expressing plasmid and empty plasmid into LoVo colon cancer cells and established two cell lines:LoVo-PRL-3 and LoVo-VC.MicroRNA chipset was used to investigate the aberrant expression of some oncomiRs.Two important members of miR-17-92 family:miR-17 and miR-19a were selected.quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) was used to validate the expression of miR-17 and miR-19a in LoVo cells.RNA interference was used to knocked down STAT3 in LoVo-PRL-3 cells,after that the expression miR-17 and miR-19a were detected.Transient transfection of miR-17 and miR-19a mimic into LoVo-VC cells or transient transfection of miR-17 and miR-19a inhibitor into LoVo-PRL-3 cells were performed to evaluate the proliferation and invasive ability of these cells by cell counting kit-8 (CCK-8) proliferating assay and Transwell chamber assay.In 13 paired primary colon cancer tissues and metastatic lesions,immunohistochemistry was used to detect the expression of PRL-3 and pSTAT3 protein,while qRT-PCR was used to investigate the expression of PRL-3,miR-17 and miR-19a.Results In LoVo-PRL-3 cells,miR-17 and miR-19 were up-regulated significantly (P ＜ 0.05).Knockdown of STAT3 mRNA decreased the expression of miR-17 and miR-19.Over-expression of miR-17 and miR-19 promoted proliferation (P ＜0.05 and P ＜0.01) and invasion (P ＜0.01) of LoVo-VC cells,while knocking down of miR-17 and miR-19 inhibited proliferation and invasion of LoVo-PRL-3 cells (P ＜0.05).PRL-3 was elevated in metastatic lesions of colon cancer and positively correlated with pSTAT3,miR-17 and miR-19a.Conclusion PRL-3 promotes proliferation and invasion of colon cancer cells by up-regulation of the expression of miR-17 and miR-19a.