IL-6 诱导LNCaP 上皮间质转化（EMT）并增强细胞对抗雄 .docVIP

二甲双胍抑制IL-6诱导的LNCaP增殖及上皮间质转化 章国亮1，姜 庆1，江 军2 （400010 重庆，重庆医科大学附属第二医院泌尿外 H1619）；重庆市国际合作项目（2011GZ0047） ［通信作者］ 姜 庆，E-mail:jq001002@；江 军，E-mail: jiangjun1964@ Metformin inhibits IL-6 induced LNCaP proliferation and epithelial-mesenchymal transition Zhang Guoliang1， Jiang Qing1， Jiang Jun2 (Department of Urology , The Second Affiliated Hospital, Chongqing Medical University, Chongqing 400010;Department of Urology , Daping Hospital/Institute of Surgery Research, The Third Medical University, Chongqing 400042) [Abstratc] Objective Detecting the mechanism by which metformin inhibits prostate cancer progression.Methods lentivirus transfection was used to construct IL-6 over-expressing LNCaP stable cell line（LNCaP-IL-6） and LNCaP control cell line（LNCaP-ctr）,Then laser scanning confocal microscope and ELISA assay was used to confirm IL-6 over-expressing LNCaP cell line; morphology of the cells was observed under inverted microscope;The following experiments were carried out on the four groups:LNCaP-ctr，LNCaP-ctr with metformin，LNCaP-IL-6 and LNCaP-IL-6 with metformin.MTT assay was used to detect cell relative number,Flowcytometry was used to analyse the affection of metformin on cell cycle; wound healing assay was used to detect the invasive ability of LNCaP-IL-6 either with or without metformin. Western blot assay was used to detect E-Cadherin and TWIST expression level of LNCaP-ctr and LNCaP-IL-6，and the alteration of them after treated with metformin. Results Laser scanning confocal microscope scanning and ELISA assay attest that the construction of LNCaP-IL-6 is successful,the secretion level of IL-6 by LNCaP-IL-6 was 5000 the amount of LNCaP-ctr.MTT assay indicated that LNCaP-IL-6 relative cell number was 1.4 times the amount of LNCaP-ctr and in metformin treated group it is up to 1.6 times after cultured for 5 days，the inhibition rates of LNCaP-IL-6 and LNCaP-ctr are 0.58 and 0.65 respectively. Flowcytometry results indicated that metformin a