Effects Of SMAD3 Gene On Biological Functions Of Colorectal Cancer Cells

Objective:SMAD3 protein is a member of the SMADs protein family,which has been proven to play a role in inhibiting or promoting cancer in the occurrence and progression of some human solid tumors.However,in colorectal cancer(CRC),research on SMAD3 is very limited.Therefore,this experiment aims to explore the expression and growth and metastasis of SMAD3 in CRC cells in vitro,and preliminarily explore the basic mechanism of its action.Methods:The lentivirus system was used to infect human colorectal cancer cell lines in order to ensure the stability of SMAD3 CRC cell lines’ knockdown and overexpression,Utilizing qRT-PCR and Western Blot,the construction efficiency is validated through the utilization of the cell counting kit-8(CCK-8)growth test,scratch test,Transwell invasion test,and migration test.The detection of SMAD3’s effect on the biological operations of CRC cells was made through a cloning test,prompting a discussion of SMAD3’s potential involvement in the infiltration and relocation of CRC.Employment of a Western blot was conducted to determine the effect of SMAD3 on the manifestation of EMT-associated.Results:Successfully constructed stable overexpression cell models of SMAD3(RKO-SMAD3,Lo Vo-SMAD3 and their controls RKO-Control,Stable knockdown cell models of SMAD3(SW0480-sh SMAD3,DLD1-sh SMAD3 and their controls SW480-sh Control,DLD1-sh Control)are Lo Vo-Control.Verification of SMAD3’s overexpression and knockdown efficacy at the protein and RNA levels in each cell model group was achieved through Western blotting and qRT-PCR,respectively.Experiments of CCK8,Transwell migration and invasion,and plate cloning revealed that in the SMAD3 stable knockdown group,cell proliferation,cell migration,and These results suggest that the expression of SMAD3 gene is negatively correlated with cell proliferation,cell migration,invasion,and clonal formation.The expression of EMT related factors was revealed to have been altered after SMAD3 expression was decreased,with a decrease in E-Cadherin being observed in colorectal cancer cells due to a decrease in SMAD3 expression.The expression of Snail,N-Cadherin,and Vimentin proteins augmented.The expression of SMAD3 was high,yet the outcomes were contrary,implying that SMAD3 can stimulate the EMT process in colorectal cancer cells.Conclusions:SMAD3 is involved in the growth and metastasis of colorectal cancer cells.The expression of SMAD3 in colorectal cancer being abnormally low can foster the proliferation,invasion and migration of these cells.Simultaneously,it can foster the EMT procedure of colorectal cancer cells.