人宫颈癌发生中FANCF表达缺陷与其外显子甲基化的关系  

Methylation of FANCF gene plays an important role in the development of cervical cancer

作　　者：李敏[1] 王泽华[2] 

机构地区：[1]重庆医科大学附属第二医院妇产科,重庆400010 [2]华中科技大学同济医学院附属协和医院妇产科,武汉430022

出　　处：《重庆医科大学学报》2008年第4期385-389,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(30070786)

摘　　要：目的:研究FANCF基因表达缺陷在宫颈癌发生发展中的作用,分析这种差异表达与甲基化的关系,并探索去甲基化对宫颈癌细胞生长的抑制作用。方法:逆转录聚合酶链技术(RT-PCR)检测宫颈癌SiHa和Hela细胞、30例宫颈癌组织标本、48例宫颈上皮内瘤样病变(Cervical intraepithelial neoplasia,CIN)组织(CINⅠ12例、CINⅡ16例、CINⅢ20例)中FANCF基因的表达。Western Blot分析FANCF蛋白水平的表达。甲基化特异性的PCR技术(MSP)对FANCF DNA进行甲基化分析。以每例标本相应正常宫颈组织作为对照。采用脱甲基化试剂5-脱氧杂氮胞苷处理FANCF甲基化的细胞,四甲基偶氮唑蓝(MTT)法检测细胞增殖。结果:SiHa、Hela细胞系以及50%(15/30)宫颈癌组织中FANCF mRNA和蛋白表达均缺失。SiHa伴有FANCF DNA甲基化,宫颈癌组织中有9例FANCF DNA高度甲基化,60%(9/15)的FANCF下降可能与甲基化有关。脱甲基化处理SiHa细胞系后,FANCF基因重表达,细胞的增殖速度下降。结论:FANCF基因甲基化导致其表达缺陷可能是宫颈癌发生的一种原因,脱甲基化处理在抑制宫颈癌的发生发展中有一定的作用。Objective：To evaluate the disruption of FANCF gene by methylation in cervical cancer and analyze the relation between cervical cancer and the methylation of FANCF DNA so as to find a method to inhibit the development of cervical cancer. Methods：The level of FANCF mRNA was detected with reverse transcription polymerase chain reaction （RT-PCR） before and after treatment with 5-aza-2＇-deoxycytidine（5-Aza-CdR）. Protein was tested by Western Blot. Methylation specific PCR（MSP） was used to check whether it was methylated in exon of FANCF. Normal cervical cancer tissues were used as controls. Results：The expression of FANCF mRNA in cervical cancer cell line SiHa was absent,and so was the expression of protein. The expression of FANCF mRNA and protein was decreased in Hela cell line. After treated with（5-Aza-CdR）,SiHa expressed FANCF mRNA and protein, and time for SiHa cells to proliferate double was prolonged. Compared with the controls,the expression of FANCF mRNA in cervical cancer tissues decreased significantly or absolutely defaulted,and expression of FANCF protein also decreased（P〈0.05）. Nine cervical cancer specimens were found methylated in exon of FANCF and the rate was 30%. 60% decreasing expression was related to FANCF methylation. All the methylated tissues were expressed lower FANCF. Conclusion：Methylation of FANCF gene plays an important role in the development of cervical cancer. Discarding the methylation in FANCF gene can inhibit the development of cervical cancer.

关 键 词：宫颈癌 FANCF 甲基化 5-脱氧杂氮胞苷 

分 类 号：R711.74[医药卫生—妇产科学]