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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 of FHIT gene in cervical cancer cells and the regulation of methylation. Methods: RJC-1, SiHa, CS1213 and C4-1 cells were cultured, DNA of these cells were extracted and subjected to bisulfite modification for PCR reaction and product detection. Analysis of FHIT gene expression in cervical cancer cells and the regulation of methylation. Results: Only the methylated primers amplified the target bands in RJC-1 and CS1213 cells, which were completely methylated. The other cells were methyl-specific primers and non-methyl-specific primers co-amplified 73bp of the purpose of the band, the status of methylation heterozygosity. After the cells were treated with 5-aza-CdR, the expression of FHIT mRNA was detected by real-time quantitative PCR, showing that the expression of FHIT mRNA in various cells increased after the treatment. Conclusion: Methylation of FHIT gene is one of the important mechanisms of down-regulation of FHIT gene expression and is one of the important directions for clinical study of cervical cancer cells.