Preliminary Study On The Effect Of Overexpression Of FBXO7 Gene On The Biological Behavior Of Colorectal Cancer Cells

Objective:Colorectal cancer(CRC)is currently one of the most common malignant tumors in the world,and its incidence rate ranks third,showing a rising trend year by year.Surgical resection is still the main way to treat colorectal cancer.Postoperative supplementation with radiotherapy and chemotherapy has made significant progress in colorectal cancer treatment,but the long-term survival of patients is still poor.F-box protein 7(FBXO7)is one of the newly discovered members of the F-box protein family.The F-box family is involved in the formation of skp1-cul1-f-box(SCF)complex,which is an important part of the ubiquitination system.At present,there is not much research on FBXO7 by scholars at home and abroad,and they mainly focus on Parkinson’s disease,but less research in the field of tumor.The purpose of this experiment is to investigate the effect of overexpression of FBXO7 gene on the biological behavior of colorectal cancer cells in order to screen early biomarkers and find new therapeutic targets.Methods:1.Human colorectal cancer cell lines HCT116 and SW620 were transfected with the recombinant plasmid over-expressing FBXO7 to construct colorectal cancer cells over-expressing FBXO7.2.Real-time PCR and Western blot were used to detect the expression of FBXO7 gene at mRNA level and protein expression in colorectal cancer cells HCT116 and SW620,which overexpress FBXO7.3.CCK-8 detected the effect of FBXO7 overexpression on the proliferation of colorectal cancer cells HCT116 and SW620 in vitro.4.The cell cycle changes of HCT116 and SW620 in colorectal cancer cells with overexpression of FBXO7 were detected by flow cytometry,and the influence of overexpression of FBXO7 on expression of Cyclin D1 in colorectal cancer cells was detected by Western blot.5.The scratching test was used to detect the change of migration ability of colorectal cancer cells HCT116 and SW620 after overexpression of FBXO7.6.Transwell invasion assay was used to detect the invasion ability of colorectal cancer cells HCT116 and SW620 after overexpression of FBXO7.Results:1.The results of Real-time PCR showed that the mRNA expression level of FBXO7 in the experimental colorectal cancer cells was significantly higher than that in the control group(P﹤0.01).Western blot results showed that compared with the control group,the expression level of FBXO7 protein in the experimental group was significantly enhanced.2.CCK-8 test results showed that in two colorectal cancer cells,compared with the control group,the OD value of the colorectal cancer cells overexpressed FBXO7 in the experimental group increased at 48 h,72h and 96 h,indicating that FBXO7 can promote colorectal Proliferation of cancer cells.3.The results of flow cytometry showed that in the two colorectal cancer cells,compared with the control group,the G1 phase proportion of the experimental group cells decreased,while the S phase proportion increased.4.The results of real-time quantitative PCR showed that the expression level of Cyclin D1 mRNA in colorectal cancer cells overexpressing FBXO7 was higher than that in the control group(P﹤0.05).Western blot showed that Cyclin D1 expression increased in colorectal cancer cells after overexpression of FBXO7.5.The scratch test results showed that compared with the control group,the colorectal cancer cells of the over-expressed FBXO7 experimental group had significantly enhanced migration ability at 24 h and 48h(P﹤0.05).6.The results of the Transwell invasion experiment showed that compared with the control group,the number of colorectal cancer cells that overexpressed FBXO7 in the experimental group passing through the matrigel compartment was significantly increased(P<0.05).Conclusion:1.Transfection of over-expressing FBXO7 recombinant plasmid successfully upregulated the mRNA and protein expression levels of FBXO7 in colorectal cancer cells HCT116 and SW620.2.Overexpression of FBXO7 can enhance the proliferation,migration and invasion of colorectal cancer cells HCT116 and SW620.3.Overexpression of FBXO7 can promote the transition of colorectal cancer cells from G1 phase to S phase,promote G1 / S phase transition,and increase the expression of cyclin Cyclin D1.