The Expression And Biological Function Of PLOD1 In Colorectal Cancer

Objective:In this study,bioinformatics technology and two-dimensional liquid chromatography-mass spectrometry(LC-MS)were used to detect colorectal cancer tumor tissues and adjacent tissues to screen out differential proteins.The up-regulated PLOD1 was selected as the target protein.Bioinformatics and mass spectrometry results were verified at the tissue level through western blot(WB)and paraffin embedded section technology in immunohistochemistry.The result was verified by WB(cell sample preparation)and real-time fluorescent quantitative PCR(RT q PCR)at the cellular level and RNA level.We also observed the effect of PLOD1 with different expression levels on the biological function of colorectal cancer cells through cell function experiment,so as to explore the impact of PLOD1 on the development of colorectal cancer.Methods:1.The expression of PLOD1 in colorectal cancer was determined by referring to Timer database and GEPIA database,and the co-expressed genes of PLOD1 were obtained by Cbioportal database;the string database was searched to obtain the PLOD1 associated protein relationship network;log in to the cosmic website to obtain the gene mutation profile of PLOD1 in colorectal cancer.2.Colorectal cancer tissues from patients were tested by LC-MS 、 WB and immunohistochemical paraffin embedded sections to detect the expression of PLOD1 protein in colorectal cancer tissues.3.SW620 was cultured as the target cell line and constructed the transient cell lines up-regulated expression and down regulated expression of PLOD1,and verified the expression of PLOD1 in each group of cell lines at the cellular and gene levels by western blot and RT q PCR.The effect of PLOD1 protein expression on cell biological function was detected by cell proliferation test,and scratch test.Results:1.PLOD1 is up-regulated in multiple tumor tissues including colorectal cancer comparing with paracancerous tissues;and the strongest association was with P3H1.The most common types of PLOD1 mutations in colorectal cancer were missense and synonymous mutations.2.The differential expression profile of colorectal cancer protein established by LC-MS found that PLOD1 was up-regulated in colorectal cancer tissue,which was also verified by western blot and immunohistochemistry.3.Differential expression profile of colorectal cancer proteins established by LCMS found that PLOD1 was up-regulated in colorectal cancer tissues,and we also validated this result using Western blot and immunohistochemistry.Cell proliferation test and cell scratch test showed that overexpression of PLOD1 could promote cell proliferation and migration,and knockdown expression of PLOD1 could inhibit cell proliferation and migration.Conclusions:1.The expression of PLOD1 was up-regulated in colorectal cancer tissues comparing with paracancerous tissues.2.The up-regulated expression of PLOD1 can promote the proliferation and migration of colorectal cancer cells.PLOD1 has potential as a biomarker for colorectal cancer diagnosis.