摘要
目的:探讨5-氮杂-2'-脱氧胞苷(5-Aza-d C)对宫颈癌细胞p16和MGMT基因表达的调控作用。方法:用5-Aza-d C处理4种宫颈癌细胞(He La、Si Ha、C33A和Ca Ski)后,MSP检测抑癌基因P16和MGMT的甲基化变化,并用荧光定量PCR和Western blot检测P16和MGMT的表达情况,同时用MTT和Annexin V-FITC/PI双染检测细胞增殖和凋亡。结果 :P16和MGMT在4种宫颈癌细胞中均存在甲基化。用5-Azad C处理细胞后,甲基化程度均被逆转。5-Aza-d C抑制P16和MGMT在宫颈癌细胞中的表达。5-Aza-d C抑制宫颈癌细胞的增殖并促进其凋亡。结论:P16和MGMT虽在宫颈癌细胞中存在甲基化,但其表达程度仍然较高,推测P16和MGMT的表达调控可能还受其他因素的影响。5-Aza-d C能抑制宫颈癌细胞的生长。5-Aza-d C虽然可以使宫颈癌细胞中P16和MGMT的甲基化程度得到逆转,但对P16和MGMT的表达却是抑制的,尚需更多实验验证并发掘其原因及机制。
Objective To discuss the regulatory effect of 5-aza-2,-deoxycytidine on P16 and MGMT in cervical cancer cells. Methods After four kinds of cervical cancer cells （HeLa, SiHa, C33A and CaSki） were treated with 5-Aza-dC, MSP was used to detect the methylation variation of P16 and MGMT, and fluorogenic quantitative PCR and Western blot were employed for determination of P16 and MGMT expression. MTT and Annexin V-FITC/PI double staining were adopted for detection of cell proliferation and apoptosis. Results Both P16 and MGMT exhibited methylation in four kinds of cervical cancer cells, and after treatment with 5-Aza-dC,their methylation levels were reversed. 5-Aza-dC was able to inhibit p16 and MGMT expression in the cervical cancer cells, and can also suppress cell proliferation and promote apoptosis. Conclusions Although methylation of PI6 and MGMT are present in cervical cancer cells, their expression level was still high. Therefore, regulation of P16 and MGMT expression may be affected by other factors. 5-Aza-dC can suppress the growth of cervical cancer cells. Although 5-Aza-dC reverse the methylation levels of P16 and MGMT, it inhibits their gene expression. More experiments are needed to verify the hidden reasons and mechanisms.
作者
陈观娣
范格英
游可理
钱德英
岑坚敏
舒焰红
李志刚
CHEN Guan-di FAN Ge-ying YOU Ke-li QIAN De-ying CEN Jin-min SHU Yan-hong LI Zhi-gang.(Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou 510080 China)
出处
《实用医学杂志》
CAS
北大核心
2016年第18期2995-2999,共5页
The Journal of Practical Medicine
基金
广东省医学科研基金资助项目(编号:A2013026)
