Cytokinesis 1 Protein Regulates The Proliferation And Apoptosis In Colon Cancer

Background and Objection:China is one of the highest incidence of colon cancer in the world,and the mor-tality of colon cancer is the top five in all malignancies.The prognosis of patients with colon cancer is closely related to the stage of tumor diagnosis,but many patients have been diagnosed in the tumor advanced stage,missing the chance of radical sur-gery.The median survival time is still short and the quality of life is poor.The development of colon cancer is a very complicated process,in which the cell indefi-nite proliferation,cell migration movement,adhesion,invasion and other changes in tumor microenvironment in tumor invasion and metastasis has a very important sig-nificance.Therefore,it is necessary to explore the related genes which effect the proliferation of colon cancer and cell apoptosis.It not only helps us to deepen our understanding of the control mechanism of colon cancer,but also provides more ex-perimental basis and theoretical basis for the early diagnosis and treatment target of colon cancer.The protein regulator of cytokinesis 1,PRC 1 gene is a key protein molecule in the mitotic phase that acts on the cytoplasmic phase.There have been several studies that PRC1 can promote the cell proliferation of gastric cancer,lung cancer,liver can-cer,breast cancer and prostate cancer and can be used as a predictor of poor prognosis.However,the function of PRC 1 gene in colon cancer,the clinical prognostic value,and the effect on colon cancer cell proliferation and apoptosis are still unclear.This study is to study the expression of PRC 1 in colon cancer for the first time and to ana-lyze its correlation with clinicopathological features and survival prognosis.The effect of PRC 1 on the biological function of colon cancer cell proliferation and apop-tosis was discussed.To provide a new theoretical basis for the pathogenesis and treatment of colon cancer.This study focused on the following three main aspects:1,the expression of PRC1 in colon cancer and its correlation with clinicopathological and prognosis;2,the function of PRC 1 on the growth and reproduction of colon cancer cells.The effect of PRC 1 on the apoptosis of colon cancer cell;3,the animal experiment on the regu-lation of colon cancer proliferation.Materials and methods:1.First analyze the mRNA expression of PRC1 in public database(www.oncomine.org).The mRNA and protein expression of PRC 1 were detected by qRT-PCR,Western blot and immunohistochemistry(IHC)in colon cancer and matched normal tissues.The expression of PRC 1 was detected by IHC method in 90 colon cancer tissues and adjacent tissues.The relationship between them and clinico-pathological features and prognosis was analyzed.2.1n the three colon cancer cell lines(HCT116,SW480,HT-29)with high ex-pression of PRC1,PRC1 was knocked down by lentivirus infection and the transfection efficiency was verified by qRT-PCR and Western blot.Experimental and plate clone formation assay can detect the changes in cell growth and proliferation capacity.The changes of cell cycle and apoptosis were detected by flow cytometry,and the changes of related cycle and apoptotic protein were detected by Western blot.3.The HCT116 cell line was divided into two groups(HCT116 empty plasmid group and HCT116-shRNA-PRC1-1 cell group).The subcutaneous tumor model of nude mice was used to observe the growth rate of the two groups of subcutaneous tumor,tumor volume and the expression of related indicators.Results:1.The subject on the Oncomine public database(www.oncomine.org)found that PRC1 in a number of colon cancer research in the chip showed high expression in colon cancer in PRC1 mRNA expression is a poor prognostic factor.qRT-PCR,West-ern blot and immunohistochemistry(IHC)were used to detect the expression of mRNA and protein in cancer tissues.The expression of PRC1 was detected by IHC method in 90 colonic cancer and adjacent tissues.The results showed that PRC1 was highly expressed in colonic cancer,and the expression of PRC1 was correlated with lymph node metastasis,tumor size and TNM stage.Survival analysis showed that the survival time of patients with high expression of PRC1 was significantly shorter than that of PRC1 low expression group.Further Cox multivariate analysis showed that PRC1 overexpression could also be used as an independent factor in the prognosis of colorectal cancer.2.The expression of PRC1 was down-regulated by qRT-PCR and Western blot.The results showed that the expression of PRC1 was significantly decreased in the HCT116,SW480 and HT-29 cell lines by lentivirus.Cell proliferation function showed that the cell growth ability of HCT116,SW480 and HT-29 cells was signifi-cantly decreased by MTT assay.Platelet clone formation assay showed that the cell proliferation ability was significantly decreased after knockdown PRC1 cell cycle.The results showed that the cells in G2 phase of HCT116 and SW480 were signifi-cantly increased and G1 phase cells were decreased in the G2 phase,and the cell cycle was mainly arrested in G2 phase.(P21,P27)protein levels were significantly up-regulated in the HCT116 and SW480 cell lines,and the correlation factors(cdc25c,CyclinB1,cdc2)protein levels were significantly reduced.In HCT116 and SW480 cell lines,Annexin-V PI staining was used to detect the number of apoptotic cells in PRC1 knockdown,and the expression of caspase3 in HCT116 and SW480 cells was detected by Western blot,Procaspase3,PARP,Bcl-x1 and Bcl2 were down-regulated,while the level of apoptosis inhibitor Cleaved caspase3 and Bax in-creased.3.The growth of subcutaneous tumor in nude mice was observed after 6 days of subcutaneous injection of HCT116 cells in lentivirus infection.And with the exten-sion of time,the difference between the two groups gradually increased.Conclusion:1.PRC1 levels in the public database of colon cancer specimens showed high expression,and high expression of mRNA can be used as a predictor of poor progno-sis of colon cancer.The expression of PRC1 protein was significantly higher than that of colon cancer,and the expression of PRC1 protein could be used as an independent prognostic factor in colon cancer.The expression of PRC1 protein was significantly higher than that of colon cancer.2.Vitro experiments show that PRC1 can promote the proliferation of colon cancer cells.In colon cancer cell lines,PRC1 inhibition can lead to G2 arrest and promote apoptosis in colon cancer cell cycle.One of the mechanisms of promoting tumor proliferation is to promote the progress of cell cycle kookiness-sis and inhibi-tion of apoptosis.3.The experiment of the body into a tumor showed PRC1 can also contribute to the proliferation of colon cancer cells.