The Experiment Research Of Cinobufacini Inhibits TGF-β1-induced Epithelial-to-mesenchymal Transition In Human Colorectal Carcinoma SW480Cells

Objective:1、 We explored whether traditional Chinese medicine Cinobufacini can effectTGF-β1-induced epithelial to mesenchymal transition in human colorectal carcinomaSW480cells.2、We study the influence of cell proliferation, invasion and migration ability onTGF-β1-induced epithelial to mesenchymal transition in human colorectal carcinomaSW480cells.Methods:1、The cultured human colorectal SW480cells were divided into the following groups:(1) control group;(2) TGF-β1individual treatment group, the dose of TGF-β1is10ng/ml;(3) Different doses of Cinobufacini intervention groups (choose2.5、5、10、20、40、80mg/ml cinobufacini co-treated with TGF-β1), the dose of TGF-β1are all10ng/ml. culture for48h.2、Morphological changes were observed by inverted phase contrast microscope.3、Proliferation and viability of the cells were measured by CCK8assay.4、The ability of cell invasion and migration about control group、TGF-β1individualtreatment group and2.5、5mg/ml Cinobufacini intervention groups were detected byTranswell assay.5、The mRNA and protein expressions of E-cadherin and Vimentin about controlgroup、TGF-β1individual treatment group and2.5、5mg/ml Cinobufacini interventiongroups were detected with QRT-PCR and Western blot.Results:1、The observation of inverted phase contrast microscope indicated that TGF-β1 individual treatment group and2.5mg/ml Cinobufacini intervention group exhibitedclassical mesenchymal phenotype compared with control group, while the5、10、20、40、80mg/ml Cinobufacini intervention groups showed classical epithelial phenotype.2、CCK8assay suggested that TGF-β1individual treatment group had no differernce,compared with control group (P>0.05). Compared with TGF-β1individual treatmentgroup，there were no difference with2.5、5mg/ml Cinobufacini intervention groups oncell proliferation of SW480cells (P>0.05).While,10、20、40、80mg/ml Cinobufaciniintervention groups had significantly inhibited cell proliferation of SW480cells(P<0.05).3、Transwell assay suggested that compared with control group, the ability of invasionand migration were significantly enhanced in TGF-β1individual treatment group(P<0.01). While,2.5、5mg/ml Cinobufacini intervention groups were significantlydecrease the ability of invasion, compared with TGF-β1individual treatment group(P<0.01).4、QRT-PCR results indicated that compared with control group the expression ofE-cadherin was significantly decreased (P<0.01) and the expression of Vimentin wassignificantly increased (P<0.01) in TGF-β1individual treatment group. While,theexpression of E-cadherin was enhanced (P<0.05) and the expression of Vimentinwas decreased (P<0.05) both in2.5、5mg/ml Cinobufacini intervention groups,compared with TGF-β1individual treatment group.5、Western blot indicated that the gray value of control group、TGF-β1individualtreatment group and2.5、5mg/ml Cinobufacini intervention groups were(0.9427±0.0254、0.4098±0.0679)、(0.3629±0.0303、0.7612±0.0472)、(0.50831±0.0165、0.6304±0.0114)、(0.7194±0.0804、0.5365±0.0199), in theTGF-β1individual treatment group the expression of E-cadherin was increased(P<0.05) and the expression of Vimentin decreased (P<0.05) as compared withcontrol group. Further more, compared with TGF-β1individual treatment group2.5、5mg/ml Cinobufacini intervention groups significantly enhanced (P<0.05) theexpression of E-cadherin and decreased (P<0.05) the expression of Vimentin. Conclusion:1、TGF-β1can induce epithelial to mesenchymal transition in colorectal carcinomaSW480cells and enhance the ability of cell invasion as well as migration.2、Cinobufacini has inhibitory effect on TGF-β1-induced epithelial to mesenchymaltransition in colorectal carcinoma SW480cells.3、Cinobufacini can inhibit the ability of proliferation as well as invasion andmigration on TGF-β1-induced SW480cells.