Effect Of JMJD2B On Biological Behavior In Human Squamous Carcinoma A431 Cells And Mechanism Research

Introduction:Squamous cell carcinoma(squamous cell carcinoma,SCC)is malignant tumor that originated epidermis or cutaneous appendages[1,2],mainly occurs in the elderly,exposure area such as head,face and hand.Its metastasis and malignant degree is low.The prognosis is poor if it growed faster or transfer[2].In recent years,many studies have found that UVB,HPV,chemical carcinogen,immunosuppression,smoking,chronic skin ulcer and burn scars are high risk factors of the SCC,but for SCC the specific pathogenesis is still not clear[1.3-6].Epigenetics is the hotspot in the study of tumor,it refers to the gene expression is heritable changed but DNA sequence did not,including DNA methylation,histone modification,chromatin remodeling,and non-coding RNA[6].Histone modification mainly includes acetylation,methylation,phosphorylation,ubiquitin and so on.Histone methylation is one of the main modified way,it is the interaction result between histone methylation and demethylation dynamic,the research showed that abnormal histone methylation can lead to gene abnormal expression,cause dysplasia and metabolic disorder[7].In recent years,the research also confirmed that histone methylation is closely related to a variety of malignant tumor development.JMJD2B is the member of the histone methylation family,it can get(trimethylated lysine 9 of histone H3,H3K9me3)specifically to demethylation[8].Previous studies have found that JMJD2B family was significantly increased expression in gastric cancer,prostate cancer,breast cancer,colorectal cancer and lung cancer[8-11],but the family relationship with the occurrence and development of SCC is not clear yet,therefore,this research is to explore JMJD2B relationship with SCC,so that providing a new method for the clinical treatment of SCC.Objective:To explore the effect of JMJD2B on biological behavior in human squamous carcinoma A431 cells and mechanism research,provide new theoretical basis for targeting therapy of SCC.Methods:1.Adopt the method of immunohistochemical and cell immunofluorescence to test the expression of JMJD2B in human squamous cell carcinoma tissue and A431 cells.2.Transfection JMJD2B siRNA to A431 cell line,the CCK-8,the clone formation test,flow cytometry and Transwell method to detect cell proliferation,the cloning,cycle distribution,apoptosis,migration and invasion situation,respectively.3.The immunocoprecipitation detected the interaction between JMJD2B and?-catenin,CyclinD1,C-MYC,JUN;Western-Blot detected expression of JMJD2B and its related protein.Results:1.The expression of JMJD2B in tumor tissue was much higher than the normal skin,and can express in the A431 cytoplasm and nucleus.2.Compared with not transfection group and transfection controls,cell proliferation,cloning,migration and invasion ability was inhibited significantly in JMJD2B siRNA transfection;the ability of cell apoptosis was increased(P<0.05),3.Immunocoprecipitation detected that there is an interaction between JMJD2B and?-catenin,CyclinD1,JUN but C-MYC;Compared with not transfection group and transfection controls,Western Blot detection ?-catenin,CyclinDl,JUN protein expression in JMJD2B siRNA transfection group was significantly reduced,.Conclusion:1.JMJD2B was highly expressed in squamous cell carcinoma and A431 cell line.2.JMJD2B siRNA can promote A431 cells apoptosis,but also inhibits the proliferation of tumor cells,cloning,migration and invasion ability and JMJD2B related protein expression.3.There is an interaction between JMJD2B and ?-catenin,CyclinD1,JUN;specifically inhibit the expression of JMJD2B,?-catenin,CyclinD1,JUN protein expression were also reduced.