TGF-β1 induced up-regulation of LMO1 drives epithelial to mesenchymal transition and metastasis in human gastric cancer MKN28 cells
10.3969/j.issn.1671-8348.2018.11.004
- VernacularTitle:TGF-β1促进人胃癌MKN28细胞上皮-间质转化和转移的研究
- Author:
Yun SUN
1
;
Guojuan MA
;
Xiaojie HU
;
Xiangyun YIN
;
Yanhui PENG
Author Information
1. 河北省人民医院普外四科
- Keywords:
stomach neoplasms;
the transforming growth factor-β1;
LMO1;
epithelial to mesenchymal transition;
metastasis
- From:
Chongqing Medicine
2018;47(11):1444-1448
- CountryChina
- Language:Chinese
-
Abstract:
Objective TGF-β1 can promote EMT,then strengthen the invasion and metastasis ability of cancer ceils.However,the mechanism for TGF-β1 in gastric cancer still keeps unclear.Aim of this study was to investigate the expression of epithelial to mesenchymal transition (EMT)marker,LMO1 and metastasis related genes on the human gastric cancer cell cell line MKN28 treated with TGF-β1,and test whether down-regulate LMO1 expression can affect the pro-EMT and pro-metastatic roles of TGF-β1 in MKN28 cells.Methods Primary human gastric cancer cell line MKN28 was cultured in vitro.Cells were treated with TGF-β1 to induce cells to undergone EMT.Cells were divided into four groups:control group (5 ％ BSA),TGF-β1 induced group (10 μg/L),negative transfect group (TGF-β1 +negative transfect siRNA),and LMO1-siRNA transfect group (TGF-β1+ LMO1-siRNA).Real time-PCR and Western blot was used to examine the difference of EMT marker (E-cadherin and N-cadherin),LMO1 and metastasis related genes (MMP-9 and VEGF)expression.Transwell assays were performed to identify the differences and changes of invasive and metastatic ability in gastric cancer cell line MKN28.Western blot was used to examine the expression levels of MMP-9 and VEGF.Results TGF-β1 stimulation induced classical EMT morphological change,as was confirmed by E-cadherin decrease and N-cadherin,LMO1,MMP-9,VEGF increase (P＜0.01).Accompanied with the EMT,cell invasion and migration ability was markedly increased (P＜0.01).However,Down-regulation of LMO1 expression reversed the pro-migratory effect of TGF-β1 to a great degree (P＜0.01).Conclusion LMO1 played a central role in coordinating TGF-β1 induced EMT and pro-migratory effects in gastric cancer MKN28 cells.Using siRNA to downregulate the expression of LMO1 can inhibit the invasion and metastasis ability of gastric cancer MKN28 cells.
