Effect Of ECM Protein SRPX2 On The Biological Behavior Of Colon Cancer Cell Line SW480

Objective The aim of this study was to investigate the expression of SRPX2 in colorectal tissues andconfirm the effect of SRPX2 on the biological behavior of colon cancer cell line SW480.Methods The expressions of SRPX2 in colon cancer tissue microarray (TMA) including 31 pairing cancer tissue and adjacent normal tissue were examined with IHC.The total RNA extracted from HCT116 cells amplified the cDNA sequence of SRPX2 using RT-PCR which was identified by 1%AGE and DNA sequencing. Sequentially the cDNA sequence was inserted into the T-vector pMD19 and eukaryotic expression vector pcDNA 3.1(+) which was identified by double digestion and DNA sequencing. The constructed plasmid was transienly transfected into SW480 using Lipofectamine 2000. The expression levels of SRPX2 mRNA and protein were detected by real-time PCR and Western blot, respectively.The impact of SRPX2 on behavior of colorectal cancer cells and possible mechanism was explored using gene transfection. The proliferation of cells were detected by using MTT assay at 24,48,72 hours after tansfection respectively.Transwell assay was adopted to determine migration and invasion ability of SW480.Result In colon cancer TMA, IHC showed that the expression of SRPX2 was significantly higher in colon cancer than in paired noncancerous tissue (p<0.001). The eukaryotic expression vector pcDNA 3.1-SRPX2 was constructed successfully and verified by double digestion and DNA sequencing.The constructed vector could express mature SRPX2 protein correctly in colon cancer cell line SW480. MTT suggested that the proliferation of SW480 cells in the pcDNA 3.1-SRPX2 transfected group was significantly higher than that in the NC group at respective time of 48,72hours (p=0.003,p= 0.017) transwell with or without matrigel showed that overexpression of SRPX2 significantly increased migratory and invasive numbers of SW480 cells(p=0.003,p=0.001). Conclusion SRPX2 protein is overexpressed in colon cancer tissues. Up-regulation of endogenous SRPX2 in SW480 cells significantly promoted the proliferation migration and invasion of SW480 cells. These findings indicate that overexpressed SRPX2 exerts an oncogenic role in colorectal cancer.