The Molecular Mechanism Of PCBP2 Regulating Colorectal Cancer Cell

Objective: The purpose of this study is to analyze the expression of PCBP2 in colorectal cancer and adjacent normal tissues by IHC Whether the expression of PCBP2 in colorectal cancer is higher than that in normal tissues,and whether the expression of PCBP2 in colorectal cancer cells and lung cancer tissues and the relationship between PCBP2 and clinical parameters are clear.In the next part,we will use HCT-116 and HCT-15 cells to confirm whether PCBP2 participates in the growth,proliferation,metastasis and apoptosis of colorectal cancer cells through cell biology and molecular biology experiments And its molecular mechanism.Methods:1.50 cases of colorectal cancer from the the first Affiliated Hospital of Anhui Medical University were selected.2.Immunohistochemistry was used to detect the expression of PCBP2 in colorectal cancer.3.The expression of PCBP 2 mRNA in HCT-116 and HCT-15 colorectal cancer cells was detected by RT-PCR.4.The protein expression of PCBP 2 in HCT-116 and HCT-15 colorectal cancer cells transfected with siPCBP2 was detected by WB.5.The proliferation of HCT-116 and HCT-15 colorectal cancer cells after si PCBP 2 was detected by soft agar colony formation assay.6.The cell viability of HCT-116 and HCT-15 colorectal cancer cells transfected with siPCBP2 was detected by CCK8 kit.7.Cell migration and invasiveness of HCT-116 and HCT-15 colorectal cancer cells transfected with siPCBP2 were detected by cell Transwell test.8.The apoptosis of HCT-116 and HCT-15 cells was detected by Annexin V / PI kit after transfection of siPCBP2.The apoptotic cells were recorded by fluorescence microscopy.Results:1.The results of immunohistochemistry showed that the expression level of PCBP2 was significantly higher in colorectal cancer patients than in normal tissues(P = 0.001),and the positive rate of PCBP 2 was 76% in colorectal cancer patients,and 40% in normal tissues(P =0.031)was related to clinical stage(P = 0.045),postoperative recurrence(P= 0.039)and total survival rate(P = 0.037).2.The results of protein imprinting showed that the expression level of PCBP 2 in HCT-116 and HCT-15 colorectal cancer cell lines transfected with siPCBP2 was significantly lower than that in NC control group.3.RT-PCR showed that the m RNA expression level of PCBP2 was significantly reduced after transfection of siPCBP2 in HCT-116 and HCT-15 colorectal cancer cell lines,and we found that the m RNA expression level of Cyclin D1 was also significantly reduced.4.CCK8 test after siPCBP2 in HCT-116 and HCT-15 colorectal cancer cell lines,the results showed that the cell viability of the two colorectal cancer cells was significantly inhibited.The cell viability of the experimental group after siPCBP2 transfection was significantly weaker than that of the NC group,and the OD value also showed that the cell growth of the experimental group was significantly faster than that of the NC group.5.The results of soft agar colony forming experiment showed that after siPCBP2 was transfected into HCT-116 and HCT-15,the colony forming of colorectal cancer cells in experimental group was significantly less than that in NC group.6.The results of cell migration showed that the number of cell migration in the experimental group was significantly less than that in the NC group after siPCBP2 was transfected into HCT-116 and HCT-15 colorectal cancer cell lines.7.The results of cell invasion experiment showed that after siPCBP2 was transfected into HCT-116 and HCT-15 cell lines,the number of cell invasion in the experimental group was significantly less than that in the NC group.8.The results of apoptosis showed that: after siPCBP2 was transfected into HCT-116 and HCT-15,the apoptosis of colorectal cancer cells in the experimental group was significantly higher than that in the NC group.Conclusions:1.The expression level of PCBP2 in colorectal cancer patients was significantly higher than that in normal tissues,suggesting that PCBP 2 is closely related to the occurrence,development,lymph node metastasis,recurrence free survival rate and prognosis of colorectal cancer.2.After siPCBP2 transfection,colorectal cancer cell viability,colorectal cancer cell growth,colorectal cancer cell clone formation,colorectal cancer cell migration and invasion were significantly inhibited,HCT-116 and HCT-15 colorectal cancer cell apoptosis increased,indicating that PCBP2 promotes the occurrence,development,metastasis and cell proliferation of colorectal cancer cells.