The Regulatory Effect Of Nuclear Protein PCNP On The Growth Of Human Thyroid Cancer Cells

BackgroundThe thyroid gland has important human endocrine functions.Thyroid cancer is an endocrine gland tumor.His pathogenesis is mostly related to the following reasons,such as geography,chemicals,and endocrine.After comprehensive clinical treatment including surgical resection,endocrine therapy and radionuclide therapy,well-differentiated thyroid papillary carcinoma and follicular carcinoma can obtain a good prognosis,but poorly differentiated thyroid cancers such as medullary thyroid carcinoma and undifferentiated carcinoma The response to existing clinical treatments is poor and the prognosis is still poor.The most important biological characteristics of malignant tumors are migration and invasion.This is not only a serious threat to human health,but also poses challenges that pathologists,surgeons and oncologists cannot ignore.Therefore,this study used the human thyroid cancer papillary cell line TPC-1 and the undifferentiated cancer cell line ARO to study the regulation of human thyroid cancer cell growth.PCNP(PEST-containing nuclear protein)is a protein ligase capable of ubiquitination.It is located in the nucleus and has PEST sequence protein(proline(P),glutamic acid(E),serine(S),and threonine(T)).PCNP can regulate the following life activities,such as cell proliferation,differentiation,cell cycle and signal transmission.PCNP was first discovered by Japanese research.PCNP,as a substrate of ubiquitin ligase NIRF(Np95/ICBP90-like RING finger protein),together with it forms a new signal transduction pathwayPCNP is ubiquitous in a variety of tissues.We have found through big data analysis that PCNP presents highly expressed protein levels in most normal tissues.However,the expression level of PCNP protein in corresponding tumor tissues is significantly reduced.PCNP expression is less in liver and skin tissues,but PCNP protein expression is relatively increased in ovarian cancer,thyroid cancer and skin cancer.Therefore,we speculate that PCNP may be related to the occurrence of thyroid cancer.In order to explore the role of PCNP in the development of thyroid cancer,this topic discusses the expression of PCNP in human thyroid cancer and its role in regulating the growth of human thyroid cancer cellsPurposeWe study the regulatory effect of nuclear protein PCNP on the growth of human thyroid cancer cellsMethodsThe expression of PCNP in human thyroid cancer was detected by constructing plasmids,transfection,cell passase,RT-PCR,qPCR,Western blot,EDU,MTS,protein extraction,RNA extraction,scratching,Transwell,soft agar colony formation,Invasion,TUNEL staining,plate cloning,tumor formation in nude mice.such as tumor and mouse identification explore the effects of PCNP on apoptosis,proliferation,migration and invasion of thyroid cancer cells from three aspects:cell model,animal model and molecular mechanism.If PCNP can really weaken the ability of cancer cells to grow,then it is very important for cancer treatment.Results1.Immunohistochemistry and Western blot results show that the expression of PCNP in thyroid cancer tissues is higher than that in adjacent tissues;2.human thyroid cancer cell PCNP overexpression and knockdown stable transfected cells were successfully constructed;3.MTS,EDU,and plate cloning experiments showed that the cell proliferation in the PCNP overexpression group was slower than in the knockdown group;4.The results of migration and scratch experiments showed that the metastasis ability of thyroid cancer cells in the PCNP overexpression group was decreased.while that of the knockout group was enhanced.5.Invasion and soft agar experiments showed that the cell invasion ability of the PCNP overexpression group decreased,while the silence group increased the cell invasion ability6.The results of Tunel apoptosis staining showed that overexpression of PCNP can accelerate the apoptosis of human thyroid cancer cells,and silence can accelerate the proliferation of human thyroid cancer cells;7.Western blot detection of protein expression of MAPK and Wnt/?-Catenin signaling pathway related molecules.The results showed that the amount of Cleaved PARP and C.leaved Caspase 3 protein in the PCNP overexpression group increased,and the amount of phosphorylated Erk,JNK and P38 protein also increased,The amount of Wnt3a protein decreased,the amount of p-GSK-3 ? and p-?-catenin protein decreased;after PCNP knockdown,the expression of Erk,JNK and P38 phosphorylation levels decreased,and the expression of GSK-3? and ?-catenin phosphorylation levels increased,Therefore,PCNP nuclear protein may regulate the apoptosis of human thyroid cancer cells through MAPK and Wnt/?-Catenin signal transduction pathways:8.Western blot analysis of cyclin expression showed that the expression of cyclinE,cyclinD1,Cdk2 and Cdk4 in the PCNP overexpression group decreased,and the expression of Cip/p21 and Kip/p27 increased,while the PCNP silence group had the opposite9.In nude mice tumorigenesis experiments,it was found that the tumor weight and volume of the PC.NP high expression group were lower than that of the control group,the positive rates of KI67,CD31 and Beclin-1 were less than that of the control group,and the staining results of Cleaned caspase 3 and P21 showed the opposite trend.ConclusionThe nucleoprotein PCNP inhibits the proliferation,migration,invasion and apoptosis of human thyroid cancer cells and may regulate apoptosis through MAPK and Wnt/?-Catenin pathway,thereby inhibiting the development of human thyroid cancer.