EZH2蛋白在非小细胞肺癌顺铂耐药中的作用及其机制研究  

Research on the role of EZH2 protein in mechanism of cisplatin resistance in non-small cell lung cancer

作　　者：齐慧[1] 呼群[2] 苏乌云[2] 贾永峰[3] 陈连香[1] 曹冉华[2] 乌日罕[2] QI Hui;HU Qun;SU Wuyun;JIA Yongfeng;CHEN Lianxiang;CAO Ranhua;WU Rihan(Department of Hematology,Affiliated Hospital of Inner Mongolia Medical University,Hohhot Inner Mongolia 010050,China;Department of Oncology,Affiliated Hospital of Inner Mongolia Medical University,Hohhot Inner Mongolia 010050,China;Department of Pathology,Affliated Hospital of Inner Mongolia Medical University,Hohhot Inner Mongolia 010050,China)

机构地区：[1]内蒙古医科大学附属医院血液内科,内蒙古呼和浩特010050 [2]内蒙古医科大学附属医院肿瘤内科,内蒙古呼和浩特010050 [3]内蒙古医科大学附属医院病理科,内蒙古呼和浩特010050

出　　处：《转化医学杂志》2020年第5期257-261,275,共6页Translational Medicine Journal

基　　金：国家自然科学基金(81160253);内蒙古自治区肿瘤生物治疗协同创新培育中心项目(1619002010);内蒙古自然科学基金(2017MS0830)。

摘　　要：目的探讨Zeste基因增强子同源物2(enhancer of zeste homolog 2,EZH2)蛋白在非小细胞肺癌对顺铂耐药中的作用及其调控机制。方法采用实时荧光定量PCR(real-time quantitative PCR,qRT-PCR)检测人肺腺癌细胞株A549及其顺铂耐药株A549/DDP中EZH2、P21、Puma和Bad的mRNA表达差异。采用RNA干扰技术抑制细胞中的EZH2蛋白表达。通过细胞毒性实验检测细胞耐药性的改变。qRT-PCR检测抑制EZH2蛋白表达后A549/DDP细胞中P21、Puma和Bad的表达变化。流式细胞仪检测细胞周期的分布和变化。结果EZH2在A549/DDP中呈高表达。抑制EZH2蛋白表达后,顺铂对A549/DDP的IC50由(34.57±3.70)μmol/L降低至(18.91±2.07)μmol/L(P<0.05);抑制EZH2蛋白表达后,A549/DDP中P21、Puma和Bad的mRNA表达均上调;抑制A549/DDP细胞EZH2蛋白表达后,G0/G1期细胞比例下降,G2/M期细胞增多(P<0.05),细胞发生G2/M期阻滞。结论EZH2蛋白参与介导了A549/DDP对顺铂的耐药;EZH2蛋白可能通过调控与细胞凋亡相关基因相关的凋亡通路诱导A549/DDP对顺铂的耐药;EZH2蛋白可能通过抑制P21表达、促进细胞周期进展及肿瘤增殖能力,诱导非小细胞肺癌对顺铂产生耐药。Objective To investigate the role and regulatory mechanism of zeste homolog 2( EZH2) protein in cisplatin resistance in non-small cell lung cancer. Methods The expression difference of EZH2,P21,Bad and Puma mRNA between A549 and A549/DDP was detected by realtime quantitative PCR. siRNA was used for silencing gene expression of EZH2. Assessment of chemoresistance to cisplatin in A549/DDP cells was performed evaluated by CCK-8 assay. The expression of P21,Bad and Puma mRNA in A549/DDP of EZH2 silenced was detected by real-time quantitative PCR. Cell cycle was analyzed by flow cytometry. Results Expression of EZH2 mRNA was overexpressed in A549/DDP compared with A549. Silencing EZH2 expression decreased IC50 of A549/DDP from( 34. 57± 3. 70) μmol/L to( 18. 91 ± 2. 07) μmol/L( P < 0. 05). Inhibiting EZH2 protein expression up-regulated expression of P21,Puma,Bad mRNA in A549/DDP. Inhibiting EZH2 protein expression caused percentage decline of G0/G1 phase,and a rise of G2/M phase( P<0. 05).Cells was arrested in G2/M phase. Conclusion EZH2 protein is involved in mediating the cisplatin resistance to A549/DDP. EZH2 protein may regulate the pathway of apoptosis to induce A549/DDP resistance to cisplatin. EZH2 protein can inhibit expression of P21,promote cell cycle progression of tumor proliferation and enhance resistance to cisplatin in non-small cell lung cancer.

关 键 词：Zeste基因增强子同源物2蛋白 肺癌 耐药性 细胞凋亡 细胞周期 

分 类 号：R734.2[医药卫生—肿瘤]