中国药理学通报Issue(8)：1084-1089,1090,7.DOI:10.3969/j.issn.1001-1978.2014.08.010
沉默EZH2表达逆转人非小细胞肺癌顺铂耐药性
Reverse effect of silencing EZH2 expression on human cisplatin-resistant non small cell lung cancer
摘要
Abstract
Aim Toinvestigatethereverseeffectofsi-lencing EZH2 on the human cisplatin-resistant non-small cell lung cancer cell line A549/DDP and its mechanism.Methods TheexpressionofEZH2was detected by real time qPCR and Western blot. siRNA was used for silencing gene expression of EZH2 . As-sessment of proliferation and chemoresistance to cispla-tin in A549/DDP cells was evaluated by MTT assay. Cell cycle and apoptosis were analyzed by flow cytome-try. Cell senescence was assessed by ß- galactosidase dyeing. The expression of H3K27me3, P14ARF, P16INK4a, P53, P21, CDK1,CDK2, Rb and E2F1 was determinedbyWesternblot.Results Theexpressions of EZH2 mRNA and protein were significantly elevated in A549/DDP cells compared with A549 cells. Silen-cing EZH2 expression resensitized A549/DDP cells to cisplatin [ IC50 from ( 41. 2 ± 4. 3 ) μmol · L-1 to (19. 4 ± 3. 3)μmol·L-1, P<0. 01], and also caused a rise of G0/G1 phase from (0. 53 ± 0. 08) to (0. 84 ± 0. 09 ) ( P<0. 01 ) , but there was not a marked varia-tion of apoptotic rate. EZH2 knockdown induced obvi-ous senescence phenotype in A549/DDP cells. EZH2-siRNA could also down-regulate the expression of CDK1 , CDK2 , H3 K27 me3 and E2 F1 , while it up-regulated the expression of P14ARF, P16INK4a, P53, P21andRbinA549/DDPcells.Conclusions Silen-cing EZH2 by RNA interference could induce cell se-nescence and resensitize A549/DDP cells to cisplatin. The mechanism of resistance reversal is associated with INK4a/ARF/Rb senescence pathway.关键词
肺癌/耐药性/siRNA/EZH2/衰老/细胞周期Key words
lung cancer/drug-resistance/siRNA/EZH2/cell senescence/cell cycle分类
医药卫生引用本文复制引用
饶进军,何关生,毛楠,钟小懿,吕琳,王文雅..沉默EZH2表达逆转人非小细胞肺癌顺铂耐药性[J].中国药理学通报,2014,(8):1084-1089,1090,7.基金项目
广东省教育部产学研结合项目(No 2012B091100465) （No 2012B091100465）
广东省自然科学基金资助项目(No S2012010009368) （No S2012010009368）
