The Role Of Ese-3 Expression In The Proliferation Of Human Colorectal Adenocarcinoma Cell Line HCT15

BackgroundThe morbidity of colorectal cancer gradually increased in recent years. And patients with concealed pathogenesis often lose the optimal opportunity of surgical therapy when colorectal cancer was diagnosed at terminal-stage. Therefore, most of those patients have poor prognosis even after multimodality therapy(surgical therapy, radiotherapy and chemotherapy). Molecular targeted drugs has a good advantage in clinical practice, thus,looking for a new target molecules and researching target molecular mechanism have become a hope for treatment in the colorectal cancer.Recent studies have demonstrated that Ese-3 is deficient/down-regulated in a variety of malignant tumors and is closely associated with tumor development and chemoresistance. So it could be used as a new target for tumor biotherapy. However, the role of Ese-3 in colorectal cancer has rarely been reported.ObjectiveThe expression of Ese-3 in para-carcinoma tissue and colorectal cancer were identified through immunohistochemical methods; moreover, lentivirus-mediated biological transfection technique was used to build a stable expression of Ese-3 gene in HCT15 cell line in order to observe the changes of cell in tumor proliferation rate, colony forming ability and cell cycle. All these would provide the foundation for theory and experiment to explore Ese-3 as a new target of colorectal cancer therapy.MethodsThe expression of Ese-3 in para-carcinoma tissue and colorectal cancer were identified through immunohistochemical methods; Western-Blot was used to detect Ese-3protein expression in cell level at colorectal cancer cell line and normal colonic epithelium cell; through realtime-q PCR, the specificity of Ese-3 amplification product in normal colonic epithelium cell was analyzed; lentivirus-mediated biological transfection technique was used to build a stable expression of Ese-3 gene in HCT15 cell line and its relative controls; Cytometry,plate colony formation assay, flow cytometer were taken to observe the changes in tumor proliferation rate, colony forming ability and cell cycle of HCT15 cell after Ese-3 overexpression.Results(1) Immunohistochemical staining results showed that the expression of Ese-3protein was different between colorectal cancer and adjacent tissues. Ese-3 was positive in normal tissue and staining particles mainly located in the nucleus.However, Ese-3 was negative in colorectal cancer tissue.(2) Compared to normal colonic epithelial cells, the expression of Ese-3 in the colon cancer cells was more significant by Western blot.(3) Quantitative real-time polymerase chain reaction was performed for the detection of Ese-3 expression. Results of real-time PCR showed that the abundance Ese-3in normal colonic epithelial cells was moderate..(4) Cytometry assay showed that the cell growth rate of HCT15-Ese-3 was lower than that of HCT15-NC. And these results demonstrated that increase of Ese-3inhibited HCT15 cell proliferation.(5) Plate colony formation assay revealed that colony formation decreased in HCT15-Ese-3 compared with HCT15-NC.(6) The percentage of G0/G1 phase in HCT15-Ese-3 increased compared with HCT15-NC(62.73% vs48.60%), and S phase decreased(21.72% vs 32.67%),P<0.05.ConclusionIn this experiment demonstrates that Ese-3 is deficient/down-regulated in colorectal cancer.Our findings show that Ese-3 could decrease HCT15 cell proliferation,inhibit HCT15 cell colony formation, arrest HCT15 cell in G0/G1 phase and inhibit HCT15 cell into S phase. These will suggest the possibility to take Ese-3 as a potential target of the biotherapy of colorectal cancer.