Role of 5'-CpG island hypermethylation of the FHIT gene in cervical carcinoma

Abstract

Objective: The abnormal expression of fragile histidine triad (FHIT) gene has been frequently reported in a variety of epithelial malignancies including cervical carcinoma. Furthermore, in a recent study it was proposed that transcriptional inactivation of FHIT, as a consequence of aberrant 5'-CpG island methylation, plays an important role in the carcinogenesis of human cervical carcinoma. The authors sought to determine whether abnormal FHIT transcription occurs in human cervical carcinoma, and if so, whether this abnormal expression is associated with aberrant 5'-CpG island methylation. In addition, the clinical significance of FHIT inactivation was investigated in Korean women with cervical cancer. Methods: To examine for abnormal transcripts of the FHIT gene, quantitative RT-PCR, genomic DNA-PCR and nonisotopic RT-PCR-SSCP analysis were performed using the standard method. The methylation status was determined by methylation specific PCR and bisulfite DNA sequencing. Results: The FHIT gene was down-regulated in 15 of 58 (25.9%) cervical carcinomas. FHIT promoter hypermethylation was detected in 15 of 15 (100%) abnormally expression in cervical carcinomas. Bisulfite DNA sequencing confirmed these findings and a significant correlation was found between CpG site hypermethylation and low FHIT expression. However, no significant correlation was found between reduced FHIT expression and clinicopathological characteristics. Conclusion: In this study, FHIT inactivation in cervical cancer was found to be strongly correlated with 5'-CpG island hypermethylation rather than a genetic alteration. Furthermore, no significant relation was found between a lack of FHIT expression and the prognostic factors of cervical cancer in our Korean cohort.

Combined Effects Methylation of FHIT, RASSF1A and RARβ Genes on Non-Small Cell Lung Cancer in the Chinese Population

“…Although the aberrant promoter region methylation was a common epigenetic event with lung cancer, combined detection of these genes may vastly improve the positive detection rate of tumors or prognostic judgment Dumitrescu et al, 2012;Vo et al, 2013;Al-Temaimi et al, 2013;Zuo et al, 2013). Several studies showed separately that methylation of CpG islands of FHIT, RASSF1A and RARβ genes has a significant role in the process of lung cancer (range, 20-90%, depending on tumor type), this figure identified the these genes as the most commonly modified gene in human cancers (Ki et al, 2008;Neyaz et al, 2008;Jeong et al, 2013;Liu et al, 2013;Al-Temaimi et al, 2013;Zuo et al, 2013). however, there are only a few studies reporting hypermethylation of combined FHIT, RASSF1A and RARβ genes together in cancers, especially in non-small cell lung cancer (Li et al, 2012;Zuo et al, 2013;Mengxi et al, 2013;Ko et al, 2013).…”

“…Methylation is major epigenetic modification in mammal, and changes in methylation patterns play a key role in tumor genesis with humans (Virmani et al, 2000;Dumitrescu et al, 2012;Fiolka et al, 2013). In particular, the promoter CpG island hypermethylation is closely related between inactivation and silencing, resulting in tumor suppressor loss of gene expression and X-chromosome inactivationmay, and affect development of carcinogenesis (Ki et al, 2008;Fiolka et al, 2013;Al-Temaimi et al, 2013). The aberrant promoter region methylation of tumor suppressor genes is associated with mechanism for carcinogenesis (Grote et al, 2006;Neyaz et al, 2008;Dumitrescu et al, 2012).…”

“…The FHIT, RARβ and RASSF1A genes were scientific researched correlating the development in human lung, gastrointestinal, liver and cervical cancer, as well as other malignant tumors as antioncogenes (Virmani et al, 2000;Neyaz et al, 2008;Ki et al, 2008;Deng et al, 2012;Li et al, 2012;Jeong et al, 2013;Liu et al, 2013;Vo et al, 2013). Although the aberrant promoter region methylation was a common epigenetic event with lung cancer, combined detection of these genes may vastly improve the positive detection rate of tumors or prognostic judgment Dumitrescu et al, 2012;Vo et al, 2013;Al-Temaimi et al, 2013;Zuo et al, 2013).…”

“…The Fragile histidine triad (FHIT) gene located on chromosome 3 at band p14.2 (3p14.2) (Ki et al, 2008;Jeong et al, 2013;Liu et al, 2013). The Ras association domain family 1 A (RASSF1A) gene located on chromosome 3 at band p21.3 (3p21.3) (Neyaz et al, 2008;Liu et al, 2013;Vo et al, 2013), and retinoic acid receptor beta (RARβ) is to the p24 band of chromosome 3 (Virmani et al, 2000;Li et al, 2012).…”

“…Three sets of primers were derived from several reports, using to amplify methylated and unmethylated alleles (Virmani et al, 2000;Ki et al, 2008;Li et al, 2012;Jeong et al, 2013). The PCR conditions of FHIT gene were 35 cycles of 94˚C for 45sec, 70˚C for 30sec and 72˚C for60sec, followed by 72˚C for 8min; The PCR conditions of RASSF1A gene were 35 cycles of 94˚C for 45sec, 62˚C for 30sec and 72˚C for60sec, followed by 72˚C for 5min; The PCR conditions of methylated RARβ gene was 35 cycles of 94˚C for 45sec, 64˚C for 30sec and 72˚C for 60sec, followed by 72˚C for 5 min; The PCR conditions of unmethylated RARβ gene was 35 cycles of 94˚C for 45sec, 59˚C for 30sec and 72˚C for 60sec, followed by 72˚C for 10min.All PCR products were analyzed on 2% agarose gel.…”