FHIT基因在宫颈癌细胞中的表达及其甲基化调控

Expression and Methylation of FHIT Gene in Cervical Cancer Cells

目的:分析FHIT基因在宫颈癌细胞中表达情况以及甲基化的调控情况。方法:对RJC-1、SiHa、CS1213以及C4-1细胞进行培养,提取这些细胞的DNA并经过亚硫酸氢盐修饰,进行PCR反应和产物的检测。分析FHIT基因在宫颈癌细胞中表达情况以及甲基化的调控情况。结果:RJC-1、CS1213细胞仅有甲基化引物扩增出了目的条带,为完全甲基化状态。其他细胞则是甲基特异性引物与非甲基特异性引物共同扩增出73bp的目的条带,其状态为甲基化杂合性。通过5-aza-CdR处理细胞后,通过实时定量PCR检测FHIT mRNA的表达,显示处理后各种细胞中的FHIT mRNA的表达升高。结论:FHIT基因的甲基化是其表达下调的重要机制之一,是临床研究宫颈癌细胞的重要方向之一。

Objective： To analyze the expression and methylation regulation of FHIT gene in cervical cancer cells. Methods： RJC-1, SiHa, CS1213 and C4-1 cells were cultured by routine method. The cells were extracted by sodium bisulfite modification. Then PCR reaction and product detection were conducted to analyze the expression and methylation regulation of FHIT gene in the cells. Results：RJC-1, CS1213 cells had the methylation primer amplified bands which was methylation status completely. SiHa and C4-1 cells were amplified by both the methyl specific primers and non-specific primers which were the heterozygous at the gene. FHIT expression under the conduction of 5-aza-CdR was detected by real-time quantitative PCR which showed that the expression of FHIT mRNA increased in the cells. Conclusion： It is suggested that the FHIT promoter methylation is an important mechanism of down-regulated expression as well as one of the important directions of clinical research on cervical cancer cells.