Experiment research of cinobufacini on TGF-β1-induced epithelial-to-mesenchymal transition in human colorectal carcinoma cells
10.3969/j.issn.1671-8348.2017.17.004
- VernacularTitle:华蟾素对TGF-β1诱导人结肠癌细胞上皮-间质化的实验研究
- Author:
Guoqiang SHEN
;
Weidong LIN
;
Xiangfang CHEN
- Keywords:
cinobufacini;
epithelial to mesenchymal transition;
transforming growth factor-beta1;
colorectal neoplasms
- From:
Chongqing Medicine
2017;46(17):2316-2319
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of cinobufacini on TGF-β1-induced epithelial-to-mesenchymal transition of human colorectal carcinoma cells in vitro.Methods The cultured colorectal carcinoma cell line(SW480) was divided into the control group,TGF-β1 (10 ng/mL) individual treatment group and co-treatment groups with TGF-β1 (10 ng/mL) + cinobufacini (2.5,5,10,20,40,80 mg/mL),which were cultured in vitro for 48 h.The proliferation of the cells were measured by CCK8 assay.The morphological changes were observed by inverted phase contrast microscope.The ability of cell invasion and migration was detected by Transwell assay.The mRNA and protein expressions of E-cadherin and Vimentin were detected with QRT-PCR and Western Blot.Results (1)Compared with the TGF-β1 (10 ng/mL) individual treatment group,TGF-β1 (10 ng/mL) + cinobufacini (10,20,40,80 mg/mL) co-treatment groups significantly had significantly proliferation inhibitory effect on SW480 (P＜0.05).(2) Compared with the normal control group,TGF-β1 individual treatment group and TGF-β1 (10 ng/mL) + cinobufacini(2.5 mg/mL) group exhibited classical mesenchymal phenotype,while the TGF-β1 (10 ng/mL) + cinobufacini (5 mg/mL) co-treatment group showed classical epithelial phenotype.(3) The ability of invasion and migration in the TGF-β1(10 ng/mL)+ cinobufacini(2.5,5 mg/mL) co-treatment group were significantly weakened compared with the TGF-β1 individual treatment group (P＜0.01).(4) QRT-PCR and Western Blot results indicated that compared with the normal control group,the Vimentin expression in the in the TGF-β1 individual treatment group was significantly increased and the E-cadherin expression was significantly decreased.Furthermore,compared with the TGF-β1 individual treatment group and control group,the Vimentin expression level in the TGF-β1 (10 ng/mL) + cinobufacini (2.5,5 mg/mL) groups was significantly decreased and E-cadherin expression was significantly increased.Conclusion Cinobufacini can inhibit TGF-β1-induced cell proliferation in human colorectal carcinoma SW480 cells,and its mechanism may be related with promoting E-cadherin expression increase,meanwhile decreasing the vimentin expression,thus inhibiting the EMT process induced by TGF-β1.
