下调JMJD2B表达对卵巢癌细胞增殖的影响  

EFFECT OF JMJD2B DOWNREGULATION ON THE PROLIFERATION OF OVARIAN CANCER CELLS

作　　者：张敏 刘晓莉[2] 张妮 雷蕾 孙超[5] ZHANG Min;LIU Xiaoli;ZHANG Ni;LEI Lei;SUN Chao(Department of Cilincal Laboratory,Jinan City People’s Hospital,Laiwu 271199,China)

机构地区：[1]济南市人民医院检验科,山东莱芜271199 [2]青岛市市立医院检验科 [3]青岛市海慈医疗集团检验科 [4]上海市东方医院妇产科 [5]青岛市市立医院高压氧科

出　　处：《青岛大学学报（医学版）》2021年第2期250-254,共5页Journal of Qingdao University(Medical Sciences)

基　　金：上海市卫生和计划生育委员会科研课题计划项目(201640246)。

摘　　要：目的探讨组蛋白去甲基化酶JMJD2B对卵巢癌细胞增殖的影响及其机制。方法应用JMJD2B siRNA和control siRNA转染人正常卵巢上皮IOSE80细胞和卵巢癌SKOV3细胞,实时荧光定量PCR和蛋白印迹法分别检测细胞中JMJD2B、环氧化酶2(COX2)mRNA和蛋白的表达水平。采用细胞克隆形成实验方法检测SKOV3细胞的增殖情况。选取20例卵巢癌病人的癌组织及癌旁正常组织,应用实时荧光定量PCR方法检测并比较两种组织中JMJD2B和COX2的mRNA表达水平。结果与IOSE80细胞比较,SKOV3细胞中JMJD2B、COX2的mRNA和蛋白水平均升高,差异有显著性(t=13.74~19.34,P<0.05)。JMJD2B siRNA转染SKOV3细胞后可以显著下调JMJD2B和COX2 mRNA的表达水平(t=4.97~7.56,P<0.05),细胞的克隆形成能力明显降低;而高表达COX2可以部分恢复细胞的克隆形成能力(F=58.23,P<0.05)。双荧光素酶实验结果显示,抑制JMJD2B的表达可以明显降低COX2的启动子活性(t=35.48,P<0.01)。人卵巢癌组织中JMJD2B和COX2的mRNA表达水平较癌旁组织显著升高(t=85.42、85.11,P<0.05),且二者呈正相关(R 2=0.983,95%CI=0.984~0.995,P<0.01)。结论降低JMJD2B水平可通过抑制COX2表达进而抑制人卵巢癌细胞的增殖。Objective To investigate the effect of the histone demethylase JMJD2B on the proliferation of ovarian cancer cells and the related mechanism.Methods JMJD2B siRNA and control siRNA were transfected into normal human ovarian epithelial IOSE80 cells and ovarian cancer SKOV3 cells,and quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression levels of JMJD2B and COX2 in these cells.Colony-forming assay was used to measure the proliferation of SKOV3 cells.Cancer tissue and normal adjacent tissue were collected from 20 patients with ovarian cancer,and quantitative real-time PCR was used to measure the mRNA expression levels of JMJD2B and COX2.Results Compared with IOSE80 cells,SKOV3 cells showed significant increases in the mRNA and protein expression levels of JMJD2B and COX2(t=13.74-19.34,P<0.05).After SKOV3 cells were transfected with JMJD2B siRNA,there were significant reductions in the mRNA expression levels of JMJD2B and COX2(t=4.97-7.56,P<0.05)and colony-forming ability,while the high expression of COX2 partially restored the colony-forming ability(F=58.23,P<0.05).The results of dual luciferase assay showed that the inhibition of JMJD2B expression significantly reduced the promoter activity of COX2(t=35.48,P<0.01).The mRNA expression levels of JMJD2B and COX2 in human ovarian cancer tissue were significantly higher than those in adjacent tissue(t=85.42,85.11;P<0.05),with positive correlation between them(R 2=0.983,95%CI=0.984 to 0.995,P<0.01).Conclusion The reduction of JMJD2B can inhibit the proliferation of human ovarian cancer cells by inhibiting the expression of COX2.

关 键 词：卵巢肿瘤 组蛋白去甲基化酶 环氧化酶2 细胞增殖 

分 类 号：R73-354[医药卫生—肿瘤]