The Experiment Research Of Hederagenin Inhibiting TGF-β1-induced Epithelial-to-mesenchymal Transition In Human Colorectal Cancer SW480 Cells

Objective:Hederagenin(HG) is pentacyclic triterpenoids which belongs to medicinal plants Clematis, now known to exist in a variety of natural plants. HG is Oleanolic acid derivatives. The literature of HG anti-tumor activity reported less,and the related research reports about oleanolic acid have been quite deeply. Due to the both have similar chemical structure,assumeing that they have some similar effects. This experiment aims to explore the transformation of epithelial-interstitial(EMT) influential mechanism about the TGF-β1-induced SW480 cell.under the role of HG From the perspective of EMT clarifying the possible mechanisms of HG inhibiting the growth of colon cancer and confronting invasion and metastasis. Providing new theoretical supports to the TCM treatment for tumor.Methods:1. The SW480 cell line was grouped as follows:①Blank control group;②10ng ml-1TGF-beta 1 induced group;③ A concentration gradient of Hederagenin intervention group(5, 10,20,40,80μg · ml-1 drug and 10 ng ml-1GF-β1 co-culture 12,24,48 h). MTT assay was used to detect the inhibition rate.2. Inverted phase contrast microscope was used to observe the cell morphology changes between the control group and the experimental group cells.3. Transwell chamber experiments was used to detect the invasion ability of cells in each group.4. Real-time PCR assay was used to observe the expressions of genes about EMT and invasion after HG acting on SW480 cells..5. Western Blot assay was used to observe the expressions of proteins about EMT and invasion after HG acting on SW480 cells.Results:1. Hederagenin inhibited the proliferation of TGF-β1-induced human colon cancer cells SW480 in a concentration and time dependent manner.2. Hederagenin could intervene the change from epithelial to mesenchymal phenotype phenotype in TGF-β1-induced SW480 cell.3. Hederagenin could inhibit the invasion and metastasis of TGF-β1-induced SW480 cell.4..After Hederagenin intervention SW480 cells, EMT and invasion related genes about N-cadherin, Vimentin, Snail, STAT3, MMP-9, MMP-14 expression decreased, the expression of E-cadherin, RECK enhancement.5. After Hederagenin intervention SW480 cells, EMT and invasion-related protein about N-cadherin, Vimentin, Snail, STAT3, MMP-9, MMP-14 expression decreased, but the expression levels of E-cadherin, RECK enhancement.Conclusions:Hederagenin could inhibit the ability of proliferation and invasion in human colorectal carcinoma cells (SW480). The mechanism may affect the expression of E-cadherin, N-cadherin, Vimentin, Snail, STAT3, MMP-9, MMP-14, RECK, causing SW480 cells EMT reversal process, thereby inhibiting the proliferation and invasion of SW480 cells.