肺癌细胞中PRDM2、PRDM5和PRDM16基因的甲基化
Methylation of PRDM2, PRDM5 and PRDM16 genes in lung cancer cells.
作者信息
Tan Shuang-Xiang, Hu Rui-Cheng, Liu Jing-Jing, Tan Yong-Li, Liu Wen-En
机构信息
Clinical Laboratory, Xiangya Hospital, Central South University Changsha, China.
Hunan Province Geriatric Hospital Changsha, China.
出版信息
Int J Clin Exp Pathol. 2014 Apr 15;7(5):2305-11. eCollection 2014.
AIMS
To investigate the changes of expression and methylation status of PRDM2, PRDM5, PRDM16 in lung cancer cells after treatment with demethylation agent.
METHODS
A549 (lung adenocarcinoma cell line), HTB-182 (lung squamous cell carcinoma cell line) and HBE (normal bronchial cell line) were treated with 5-aza-2dC. The methylation state of PRDM2, PRDM5, PRDM16 was detected by MSP. The expression of PRDM2, PRDM5, PRDM16 was detected by RT-PCR and Western blot analysis. Cell growth was detected by MTT assay.
RESULTS
5-aza-2-dC reduced the methylation of PRDM2, PRDM5, PRDM16 gene in A549 and HTB-182 cells but not in HBE cells. Consistently, 5-aza-2dC increased mRNA and protein expression of PRDM2, PRDM5, PRDM16 in A549 and HTB-182 cells but not in HBE cells. Furthermore, 5-aza-2dC inhibited the growth of A549 and HTB-182 cells but not HBE cells.
CONCLUSIONS
PRDM2, PRDM5, PRDM16 promoters are methylated and their expression is suppressed in lung cancer cells. Demethylation drug 5-aza-2dC could upregulate the expression of PRDM2, PRDM5, PRDM16 and suppress lung cancer cell growth. 5-aza-2dC has potential to be used for lung cancer therapy by epigenetic mechanism.
目的
研究去甲基化剂处理后肺癌细胞中PRDM2、PRDM5、PRDM16的表达及甲基化状态变化。
方法
用5-氮杂-2'-脱氧胞苷（5-aza-2dC）处理A549（肺腺癌细胞系）、HTB-182（肺鳞癌细胞系）和HBE（正常支气管细胞系）。通过甲基化特异性PCR（MSP）检测PRDM2、PRDM5、PRDM16的甲基化状态。通过逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹分析检测PRDM2、PRDM5、PRDM16的表达。采用MTT法检测细胞生长情况。
结果
5-aza-2dC降低了A549和HTB-182细胞中PRDM2、PRDM5、PRDM16基因的甲基化水平，但对HBE细胞无此作用。同样，5-aza-2dC增加了A549和HTB-182细胞中PRDM2、PRDM5、PRDM16的mRNA和蛋白表达，但对HBE细胞无此作用。此外，5-aza-2dC抑制了A549和HTB-182细胞的生长，但对HBE细胞无此作用。
结论
PRDM2、PRDM5、PRDM16启动子在肺癌细胞中发生甲基化，其表达受到抑制。去甲基化药物5-aza-2dC可上调PRDM2、PRDM5、PRDM16的表达并抑制肺癌细胞生长。5-aza-2dC有通过表观遗传机制用于肺癌治疗的潜力。
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