精氨酸甲基转移酶5在甲硫腺苷磷酸化酶缺陷型恶性胸膜间皮瘤中的作用研究  

Role of protein arginine methyltransferase 5 in methylthioadenosine phosphorylase deficient malignant pleural mesothelioma

作　　者：张奇 王志强 蔡华荣 江跃全 ZHANG Qi;WANG Zhiqiang;CAI Huarong;JIANG Yuequan(Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment,Cancer Hospital Affiliated to Chongqing University,Chongqing,400030)

机构地区：[1]重庆大学附属肿瘤医院,肿瘤转移与个体化诊治转化研究重庆市重点实验室,重庆400030

出　　处：《实用临床医药杂志》2021年第17期82-87,共6页Journal of Clinical Medicine in Practice

基　　金：重庆市科卫联合医学科研项目(2021MSXM214)。

摘　　要：目的探讨抑制精氨酸甲基转移酶5(PRMT5)基因表达对甲硫腺苷磷酸化酶(MTAP)缺陷型恶性胸膜间皮瘤(MPM)细胞的联合杀伤作用。方法将MPM细胞分为MTAP阳性组(REN、H28、MPP89)和MTAP缺陷组(H2591、2452、H2052),以Hap-1 MTAP KO/WT为对照。采用PRMT5 siRNA、奎纳克林(0、0.5、1.0μmol/L)处理2组细胞,72 h后采用Western blot检测PRMT5蛋白表达水平及H4R3me2s甲基化蛋白水平。结晶紫染色法测定细胞克隆增殖能力。结果PRMT5 siRNA可下调MTAP阳性细胞株及MTAP缺陷细胞株PRMT5蛋白水平,引起MTAP缺陷型MPM细胞的H4R3me2s甲基化水平明显下降,而对MTAP阳性细胞的H4R3me2s甲基化水平无明显抑制。PRMT5 siRNA抑制MTAP缺陷型细胞的克隆增殖能力,而对MTAP阳性细胞克隆无明显抑制作用,差异有统计学意义(P=0.023,n=3)。奎纳克林可下调MTAP缺陷型细胞系(H2591、H2052)中PRMT5蛋白质表达水平,引起H4R3me2s甲基化水平降低及细胞克隆增殖能力下降,而对MTAP阳性细胞系(MPP89)中PRMT5和H4R3me2s均无下调作用,对细胞增殖无抑制,差异有统计学意义(P=0.018,n=3)。结论抑制PMRT5能联合杀伤MTAP缺陷型MPM,奎纳克林对部分MTAP缺陷型MPM可产生联合杀伤作用。Objective To explore the combined killing effect of inhibiting the expression of protein arginine methyltransferase 5(PRMT5)gene on methylthioadenosine phosphorylase(MTAP)deficient malignant pleural mesothelioma(MPM)cells.Methods MPM cells were divided into MTAP positive group(REN,H28 and MPP89)and MTAP deficient group(H2591,2452 and H2052),and the Hap-1 MTAP KO/WT were used as controls.PRMT5 siRNA and quinacrine(0,0.5 and 1.0μmol/L)were used to treat cells in two groups,and the expression levels of PRMT5 protein and H4R3me2s methylated protein were detected by Western blot after 72 hours.The ability of cell clone and proliferation was determined by crystal violet staining.Results PRMT5 siRNA was able to downregulate the PRMT5 protein levels of MTAP positive cell lines and MTAP deficient cell lines,resulting in a significant decrease in the H4R3me2s methylation level of MTAP deficient MPM cells,but had no significant inhibitory effects on the H4R3me2s methylation level of MTAP positive cells.PRMT5 siRNA was able to inhibit the clonal and proliferation ability of MTAP deficient cells,but had no significant inhibitory effect on the cloning of MTAP positive cells(P=0.023,n=3).Quinacrine was able to downregulate the expression level of PRMT5 protein in MTAP deficient cell lines(H2591 and H2052),reduce the methylation level of H4R3me2s and the ability of cell clone and proliferation,but had no down-regulation effect on PRMT5 and H4R3me2s in MTAP positive cell lines(MPP89)and no inhibitory effect on cell proliferation,and there were significant differences(P=0.018,n=3).Conclusion Inhibition of PMRT5 can kill MTAP deficient MPM in combination,and quinacrine can produce combined killing effect on some MTAP deficient MPM.

关 键 词：精氨酸甲基转移酶5 甲硫腺苷磷酸化酶 恶性胸膜间皮瘤 克隆增殖能力 奎纳克林 

分 类 号：R394.2[医药卫生—医学遗传学] R734.3[医药卫生—基础医学]