Expression Of TIPE3 In Colorectal Cancer And Its Clinical Significance

Objective:To clarify the expression of tumor necrosis factor-alpha-induced protein 8-like 3(TNFAIP8L3,TIPE3)in colorectal cancer(CRC)and its relationship with clinical pathological factors and prognosis of patients.Methods:(1)We collected the total protein of HT29,LOVO,LS174T and SW480 CRC cells and NCM460 normal intestinal mucosal epithelial cells.The expression of TIPE3 in CRC cells and normal control cells was detected by Western blot.(2)The GFP-tagged and TIPE3-overexpressed lentiviral plasmid pLenti6.3/TIPE3/IRES/GFP,control empty lentiviral plasmid pLenti6.3/IRES/GFP and helper plasmids including pLP1,pLP2 and VSVG were amplified,respectively.After co-transfection with 293T cells,TIPE3-overexpressed lentivirus(LV-TIPE3)and control empty lentivirus(LV-NC)were prepared.(3)The SW480 CRC cells were infected with 50 multiplicity of infection(MOI)of LV-TIPE3 or LV-NC.The TIPE3-overexpressed SW480 transgenic cells SW480-TIPE3 and its control cells SW480-NC were obtained through selection with Blasticidin S.The expression of GFP was analyzed by fluorescence microscope and flow cytometry.The overexpression of TIPE3 in SW480-TIPE3 cells was determined by Western blot and qRT-PCR.(4)The effect of TIPE3 overexpression on migration and invasion ability of CRC cells in vitro was detected by wound healing assay and Transwell invasion assay,respectively.(5)A total of 107 pairs of CRC tumor and matched paracancerous tissues derived from 107 CRC patients were harvested and selected for preparation of human CRC tissue microarray(TMA)(107 cases).The human CRC TMA(HColA180Sull)(93 cases)was also purchased directly.The expression of TIPE3 in CRC tumor and adjacent tissues using above CRC TMAs was detected by immunohistochemistry;The correlation between expression level of TIPE3 in CRC tumor tissues and clinical pathological factors was then defined.(6)The overall survival time of CRC patients with high and low TIPE3 expression groups was followed up,and the relationship between TIPE3 expression level and prognosis of CRC patients was analyzed by The Cancer Genome Atlas(TCGA)cancer public database through Gene Expression Profiling Interactive Analysis(GEPIA).Results:(1)Compared with NCM460 human normal intestinal mucosal epithelial cells,the expression level of TIPE3 in CRC cells was increased.(2)The TIPE3-overexpressed CRC transgenic cells SW480-TIPE3 and control cells SW480-NC were established.(3)The migration and invasion ability of lentivirus-mediated TIPE3-overexpressed CRC cells SW480-TIPE3 were significantly enhanced compared with control cells SW480-NC.(4)The expression level of TIPE3 in CRC tumor tissues was significantly upregulated compared with that in control paracancerous tissues,and the expression level of TIPE3 in tumor tissues was positively correlated with malignant clinicopathological features such as lymph node metastasis and tumor stage of CRC patients.(5)The overall survival time of CRC patients in the TIPE3 high expression group was shorter than that in the TIPE3 low expression group,which was consistent with the prognosis data from GEPIA analysis of TCGA database.Conclusions:TIPE3 expression is elevated in human CRC.TIPE3 has the ability to promote the migration and invasion of CRC cells.The expression level of TIPE3 is positively correlated with lymph node metastasis and tumor stage malignant clinicopathological features in human CRC.High TIPE3 expression is also closely associated with poor prognosis of CRC patients.