Construction of a eukaryotic expression vector of TM4SF1 and its effect on migration and in-vasion of colorectal cancer cells

VernacularTitle:TM4SF1真核表达载体的构建及其对结直肠癌细胞株生物学功能的影响
Author: Dan WANG ¹ ; Na CHEN ; Man PENG ; Qiong XU ; Jun ZHOU
Author Information

1. 南方医科大学基础医学院病理学系
Keywords: transmembrane-4-L-six-family-1; migration; invasion
From: Journal of Southern Medical University 2014;(6):847-851
CountryChina
Language:Chinese
Abstract: Objective To construct a eukaryotic expression vector of transmembrane-4-L-six-family-1 (TM4SF1) gene and study its effect on the migration and invasion of colorectal cancer cells. Methods A pair of specific primers of TM4SF1 gene (GenBank: BC034145.1) was used to acquire the open reading frame of TM4SF1 by RT-PCR. The amplified sequence was ligated to a PEZ-M29 vector, which, after identification, was transiently transfected in colorectal cancer cell lines HCT116 and DLD1. Western blotting and immunocytochemistry were used to analyze the transfection efficiency, and scratch and Transwell tests were performed to analyze the changes in the migration and invasion of HCT116 and DLD1 cells after transfection. Results Cell scratch and Transwell assays revealed that transfection with the recombinant plasmid, PEZ-M29/TM4SF1, caused up-regulated expression of TM4SF1 and promoted the migration and invasion of HCT116 and DLD1 cells. Conclusion Our results demonstrated that TM4SF1 is closely related to the invasion and metastasis of colorectal cancer cells in vitro.