RNA干扰mPGES-1基因对K562/A细胞增殖及凋亡的影响  

Effects of RNA Targeting mPGES-1 on Proliferation and Apoptosis of K562/A Cells

作　　者：邱梦 聂大年[1] 谢双锋[1] 肖洁[1] 吴裕丹[1] 王秀菊[1] 杨文娟 李益清[1] QIU Meng;NIE Da-nian;XIE Shuang-feng;XIAO Jie;WU Yu-dan;WANG Xiu-ju;YANG Wen-juan;LI Yi-qing(Department of Internal Hematology,Sun Yat-Sen Memorial Hospital,Sun Yat-Sen University//Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation,Guangzhou 510120,China)

机构地区：[1]中山大学孙逸仙纪念医院血液内科//广东省恶性肿瘤表观遗传学和基因调控重点实验室,广东广州510120

出　　处：《中山大学学报（医学科学版）》2020年第2期233-242,共10页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81803783);广东省自然科学基金(2017A030313612;2016A030313270)。

摘　　要：[目的]探讨RNA干扰膜结合型前列腺素E2合酶1(mPGES-1)对阿霉素耐药的人红白血病细胞株K562/A细胞增殖、凋亡及耐药性的影响及其可能的机制.[方法]通过RNA干扰技术抑制K562/A细胞中mPGES-1表达.分组:①未处理组(K562/A),②干扰后的阴性对照组(K562/A-NC),③干扰组(K562/A-KD),④干扰后加入外源性PGE2组(K562/A-KD+PGE2);CCK-8法检测细胞活力;流式细胞术检测细胞凋亡;ELISA法检测PGE2浓度;Western blot检测蛋白水平.[结果]RNA干扰可明显下调K562/A细胞mPGES-1表达,抑制PGE2合成(P<0.0001).RNA干扰后,K562/A细胞增殖受抑,凋亡增加,对各化疗药物敏感性均有不同程度的增强(P<0.05),同时β-catenin、MDR1的表达量减少(P<0.01).外源性PGE2可逆转RNA干扰对K562/A增殖、凋亡水平、药物敏感性的影响(P<0.05),同时β-catenin、MDR1的表达上调(P<0.01);β-catenin抑制剂XAV939可浓度依赖性抑制β-catenin、MDR1蛋白的表达(P<0.05).[结论]RNA干扰mPGES-1基因可抑制K562/A细胞增殖、诱导细胞凋亡、增强细胞对化疗的敏感性,其机制与减少PGE2的合成进而下调β-catenin、MDR1的表达有关.Wnt/β-catenin通路可能参与了mPGES-1/PGE2对MDR1的调控.【Objective】To explore the effects and the possible mechanism of RNA targeting membrane-bound prostaglandin E2 synthase l(mPGES-1)on proliferation,apoptosis and drug resistance of leukemia cell line K562/A.【Methods】RNA interference was used to inhibit the expression of mPGES-1 of K562/A cells.Four groups were set up as follows:untreated group(K562/A),negative control group after interference(K562/A-NC),group after interference(K562/A-KD),and group after interference with exogenous PGE2(K562/A-KD+PGE2).Cell viability was assessed by CCK-8 assay.Cell apoptosis was analyzed by flow cytometry.Concentration of PGE2 was detected by ELISA.Proteins expression was detected by western blot.【Results】The expression of mPGES-1 in K562/A cells was significantly down-regulated and the synthesis of PGE2 decreased(P<0.0001)after RNA interference.After RNA interference,the proliferation of K562/A cells was inhibited and apoptosis increased,and the sensitivity to chemotherapy drugs was enhanced(P<0.05).Meanwhile,the expression ofβ-catenin and MDR1 was decreased(P<0.01).Exogenous PGE2 could reverse the effect of RNA interference on proliferation,apoptosis and drug sensitivity in K562/A cells(P<0.05),and up-regulate the expression ofβ-catenin and MDR1(P<0.01).XAV939,an inhibitor ofβ-catenin,could down-regulate the expression of β-catenin and MDR1 in an dose-dependent pattern in K562/A cells(P<0.05).【Conclusions】RNA interference of mPGES-1 could inhibit proliferation,induce apoptosis and reverse drug resistance in K562/A cells.The mechanism was related to reducing the synthesis of PGE2 and thus down-regulating the expression ofβ-catenin and MDR1.Wnt/β-catenin signal pathway may participate in the regulation of MDR1 by mPGES-1/PGE2.

关 键 词：MPGES-1 PGE2 白血病 耐药 Β-CATENIN 

分 类 号：R733[医药卫生—肿瘤]