5-氮杂-2’-脱氧胞苷通过反转原钙黏蛋白10表达抑制人乳腺癌细胞系MDA-MB-231的侵袭和迁移  

Effect of protocadherins 10 re-expression induced by 5-aza-2’-deoxycytidine on invasion and migration capacity of breast cancer cell line MDA-MB-231 and its mechanism

作　　者：张微 朱文斌 岳丽玲 刘立琨 ZHANG Wei;ZHU Wen-bin;YUE Li-ling;LIU Li-kun(Research Institute of Medicine and Pharmacy, Qiqihar Medical University Heilongjiang Qiqihar 161006, China;Pharmacy School , Jiamusi University y Heilongjiang Jiamusi 154007, China)

机构地区：[1]齐齐哈尔医学院医药科学研究院,黑龙江齐齐哈尔161006 [2]佳木斯大学药学院,黑龙江佳木斯154007

出　　处：《解剖学报》2019年第4期465-470,共6页Acta Anatomica Sinica

基　　金：黑龙江省自然科学基金(H2014100);齐齐哈尔市科技局项目(SGFF-201632);黑龙江省普通本科高等学校青年创新人才培养计划(UNPYSCT-2017166)

摘　　要：目的探讨5-氮杂-2’-脱氧胞苷(5-Aza-CdR)诱导抑癌基因原钙黏蛋白10(PCDH10)重新表达对人乳腺癌细胞MDA-MB-231体外侵袭迁移能力的影响并初步探讨其机制。方法体外培养人乳腺癌细胞MDA-MB-231,设置对照组和5-Aza-CdR药物处理组,分别采用反转录聚合酶链反应(RT-PCR)检测PCDH10 mRNA的表达水平;Transwell法和划痕实验检测细胞的侵袭迁移能力;Western blotting检测PCDH10、DNA甲基转移酶(DNMT) 3A、DNMT3B、核因子(NF)-κB p65和基质金属蛋白酶(MMP)-2、MMP-9蛋白表达的变化。结果 5-Aza-CdR能够反转PCDH10的mRNA和蛋白表达;PCDH10表达恢复后MDA-MB-231细胞的侵袭迁移能力受到抑制;Western blotting检测发现,MDA-MB-231细胞经5-Aza-CdR处理后DNMT3A、DNMT3B、NF-κB p65、MMP-2和MMP-9的表达下调。结论 5-Aza-CdR可抑制MDA-MB-231细胞DNMT3A和DNMT3B的表达,使抑癌基因PCDH10表达恢复,从而通过阻滞NF-κB p65的活化,下调MMP-2和MMP-9表达而抑制乳腺癌细胞的侵袭转移。Objective To investigate whether re-expression of protocadherin 10(PCDH10) induced by 5-aza-2’-deoxycytidine(5-Aza-CdR) could affect the invasion and migration of MDA-MB-231 cells,and to explore the possible mechanism. Methods Human breast cancer cell line MDA-MB-231 was cultured in vitro. Control group and 5-Aza-CdR treatment group were set up. PCDH10 mRNA expression in MDA-MB-231 cell line was determined by reverse transcription-polymerase chain reaction(RT-PCR);Transwell chamber and wound healing assay were performed to measure the invasion and migration capacity of the cells,and protein expression of PCDH10,DNA methyltransferase(DNMT) 3 A,DNMT3 B,nuclear factor(NF)-κB p65,matrix metalloproteinases(MMP)-2 and MMP-9 were detec Western blotting. Results 5-Aza-CdR could reverse the methylation status of PCDH10 gene in MDA-MB-231 cells in a dosedependent manner. Re-expression of PCDH10 significantly inhibited cell invasion and migration capacity in vitro. Western blottoing analysis revealed that the expression of DNMT3 A,DNMT3 B,NF-κB p65,MMP-2 and MMP-9 in MDA-MB-231 cells were down-regulated after exposure to 5-Aza-CdR. Conclusion Re-expression of PCDH10 significantly inhibits MDA-MB-231 invasion and migration capacity. The inhibitory effect is characterized that 5-Aza-CdR treatment downregulates DNMT3 A and DNMT3 B levels,recovers the expression of anti-oncogene PCDH10,further blocks the activation of NF-κB p65,resultsing in a decrease in the secretion of MMP-2 and MMP-9.

关 键 词：乳腺癌 原钙黏蛋白10 甲基化 侵袭 迁移 反转录聚合酶链发应 免疫印迹法 人 

分 类 号：R730.2[医药卫生—肿瘤]