Study On The Mechanism Of Over-expression Of NUDT21 In Inhibiting The Proliferation Of Cancer Cells

ObjectiveSo far,cancer remains a relatively hot topic worldwide and is one of the major factors affecting human health.In recent years,molecularly targeted therapies have received increasing attention because of their ability to precisely identify and specifically treat cancer cells while having a smaller impact on normal cells.Molecularly targeted therapeutic approaches are revolutionary in that they interfere with specific molecules to achieve the goal of stopping the growth,progression and metastasis of cancer cells.NUDT21 plays a central role in alternative cleavage of the mRNA 3’UTR,and some studies have found aberrant expression of NUDT21 in cancer cells;Cx43 is an important protein for cellular gap junctions and is associated with cancer development associated with cancer development.In this study,we explored the roles of NUDT21 and Cx43 in colon cancer cells or lung cancer cells,respectively,and explored the interactions between them in the cells,providing a basis for molecular targeting therapy.In this project,we first explored the role of NUDT21 in colon cancer.Colon cancer cells HCT-116 were used as experimental cells,and after the NUDT21 overexpression plasmid was constructed by Gibson method and transfected in HCT116 cells,the proliferation of cells was detected by CCK-8 assay and colony formation assay,and the distribution of each phase of the cell cycle was detected by flow cytometry,and it was found that NUDT21 could inhibit the proliferation of HCT-116 cells.Subsequently,the expression of related cyclin and cyclin ubiquitin E3 ligase was continued to be detected,and it was found that after overexpression of NUDT21,the expression of Cyclin D1 and Cyclin D3 was increased,and the expression of FBXW8 and FBXO31 protein was decreased,the expression of Cyclin E1 protein was decreased,the expression of Cyclin E2 protein was not significantly different,and the expression of p53 protein increased;Rb-Ser608 and Rb-Ser780 phosphorylation levels were reduced;CDK2 expression was reduced and CDK4 was not significantly changed.Then we explored the relationship between NUDT21 and Cx43 and found that:the results of immunoprecipitation assay suggested a binding relationship between the two;CCK-8 results showed that overexpression of both NUDT21 and Cx43 in A549 cells reduced the viability of cell proliferation;Western blot results showed that overexpression of NUDT21 in A549 cells increased the expression of Cyclin D1 and Cyclin D3 protein expression was increased;overexpression of Cx43 in A549 cells decreased Cyclin D1 and Cyclin D3 protein expression,while the protein expression levels of FBXO31,FBXW8,and FBXO4 were also increased;fluorescence quantitative PCR results revealed that,compared to the control,overexpression of Cx43,FBXO31,FBXW8 and the mRNA expression levels of FBXO4 were increased when compared to the control group.From the above studies,it was finally concluded that,firstly,NUDT21 has the ability to inhibit cancer cell proliferation in both lung cancer cells A549 and colon cancer cells HCT-116;secondly,although there is a binding relationship between NUDT21 protein and Cx43 protein,there may not be other relationships in the cellular mechanism pathway.