Ascl2表达抑制对结肠癌细胞上皮-间质转化的影响.pdfVIP

201341 384 Ascl2 - []Ascl2 - (EMT)Ascl2 shRNA HT-29 LS174T G418MTTAscl2 Real-time PCR Western blotting Ascl2 EMTE- MTTAscl2 72h96h HT-29LS174T (P 0.05) HT-29 LS174T HT-29 LS174T (P 0.05) Real-time PCR Western blotting HT-29 LS174T E- mRNA (P 0.05) mRNA (P 0.05) Ascl2 RNA HT-29LS174TEMT [] []R735.35 []A []0577-7402(2013)04-0292-05 * TIAN Yin, ZHU Rong, WANG Rong-quan Institute of Gastroenterology of PLA, Southwest Hospital, Third Military Medical University, Chongqing 400038, China * Corresponding author, E-mail: rongquanw@ [Abstract]ObjectiveTo investigate the regulation of epithelial-mesenchymal transition (EMT) in colon cancer cells by Achaete scute-like 2 (Ascl2) knockdown. MethodsHuman colon cancer cells HT-29 and LS174T, were transfected with shRNA- Ascl2 vector, then the transfected cells were selected with G418 and stably transfected cell lines were established. MTT assay and colony formation experiment were used to observe the effects of Ascl2 RNA interference on cell proliferation and colony formation ability. Real-time PCR and Western blotting analysis were used to detect the expression of E-cadherin and Vimentin mRNAs and proteins. ResultsMTT assay found that the cell proliferation rate of HT-29 and LS174T cells transfected with shRNA-Ascl2 vector was significantly lower than the cells transfected with shRNA-control vector and the untransfected cells 72 and 96 hours later after inoculation (P 0.05). The colony formation experiment indicated that the colony formation numbers of HT-29 and LS174T cells transfected with shRNA-Ascl2 vector was significantly less than the cells transfected with shRNA-control vector and the untransfected cells 20 days after inoculation. The expression levels (mRNA and protein) of E-