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为探讨膀胱癌相关蛋白磷酸酶2A催化亚单位(BCAPP2Ac)新基因在膀胱癌组织中的表达及其对膀胱癌细胞增殖的影响,通过合成抗原多肽免疫家兔获得抗BCAPP2Ac多克隆抗体及通过慢病毒感染方式获得稳定表达BCAPP2Ac的膀胱癌细胞.采用实时PCR及免疫组化染色方法检测BCAPP2Ac mRNA和蛋白在膀胱癌组织、癌旁组织及其它多种肿瘤组织中的表达;用细胞增殖实验检测BCAPP2Ac对膀胱癌细胞增殖的影响.实时PCR及免疫组化结果显示,BCAPP2Ac mRNA及蛋白在膀胱癌组织较癌旁组织表达明显下调;过表达BCAPP2Ac能抑制膀胱癌EJ、T24细胞的增殖,蛋白磷酸酶2A催化结构域缺失(△PP2Ac)能逆转BCAPP2Ac的增殖抑制作用.研究结果提示,新基因BCAPP2Ac能抑制膀胱癌细胞的增殖,PP2Ac结构域在其抑制细胞增殖作用中发挥重要作用.
To investigate the expression of bladder cancer-associated protein phosphatase 2A catalytic subunit (BCAPP2Ac) in bladder cancer and its effect on the proliferation of bladder cancer cells, polyclonal anti-BCAPP2Ac antibody was obtained by immunizing rabbits with synthetic antigen peptide BCAPP2Ac was detected by real-time PCR and immunohistochemistry.The expression of BCAPP2Ac mRNA and protein in bladder cancer tissues, paracancerous tissues and other tumor tissues were detected by real-time PCR and immunohistochemistry.The cell proliferation assay was used to detect the expression of BCAPP2Ac On the proliferation of bladder cancer cells.Results of real-time PCR and immunohistochemistry showed that BCAPP2Ac mRNA and protein were significantly down-regulated in paracancerous tissues of bladder cancer; over-expression of BCAPP2Ac inhibited the proliferation of bladder cancer EJ and T24 cells, and protein phosphatase 2A △ 2A2 catalytic domain deletion (△ PP2Ac) can reverse the proliferation inhibition of BCAPP2Ac.The results suggest that the new gene BCAPP2Ac can inhibit the proliferation of bladder cancer cells, PP2Ac domain plays an important role in its inhibition of cell proliferation.