Butler D, Collins C, Mabruk M, Barry Walsh C, Leader M B, Kay E W
Department of Pathology, Beaumont Hospital and Royal College of Surgeons in Ireland, Education & Research Centre, Smurfit Building, Beaumont Hospital, Dublin 9, Ireland.
J Pathol. 2000 Dec;192(4):502-10. doi: 10.1002/1096-9896(2000)9999:9999<::AID-PATH718>3.0.CO;2-H.
The fragile histidine triad (FHIT) gene encompasses the common chromosomal fragile site FRA3B. Human papilloma virus (HPV), which is the main aetiological agent in cervical cancers, has been found to be able to integrate its genes into the chromosome 3 fragile site of cultured cells, deleting a piece of DNA which includes the FHIT gene. Eighty-six microdissected archival cervical LLETZ biopsies comprising cases of cervical intraepithelial neoplasia (CIN) 1 (n=27), CIN3 (n=30) and microinvasive carcinoma (n=29) were evaluated for HPV infection and FHIT gene loss of heterozygosity (LOH). FHIT gene LOH was detected by polymerase chain reaction (PCR) using fluorescently labelled intragenic microsatellite markers D3S1300 and D3S4103. PCR products were analysed on a semi-automated DNA sequencer using Fragment Manager(trade mark) software to determine allele loss. The HPV status of the lesions was determined by PCR using generic and type-specific primers in conjunction with restriction endonuclease digestion. The results were analysed using Epi-Info and SPSS-PC statistical analysis software. Haematoxylin and eosin-stained sections from the 86 cases were profiled for six histopathological features, some of which have been previously shown to be associated with microinvasive cancer. FHIT gene LOH was found in 36% of CIN1 cases, 52% of CIN3 cases and 73% of microinvasive cases (p=0.029). HPV 16 DNA was found in 68% of CIN3 cases and 93% of microinvasive cases (p<0.001). The second most prevalent HPV type found was HPV 31, which was present in only four lesions, three of which had FHIT gene LOH. When FHIT gene LOH was evaluated versus HPV 16 and 31 infection using the chi-square test, a statistically significant relationship was found (p=0.014). FHIT gene LOH was found to be independent of the histopathological features evaluated. The finding of a statistically significant relationship between FHIT gene LOH and oncogenic HPV infection suggests a link between the integration of viral DNA and subsequent gene deletion in the progression of cervical cancer. FHIT gene anomalies may prove to be excellent markers of progression in early uterine cervical cancers.
脆性组氨酸三联体（FHIT）基因包含常见的染色体脆性位点FRA3B。人乳头瘤病毒（HPV）是宫颈癌的主要致病因子，已发现它能够将其基因整合到培养细胞的3号染色体脆性位点，缺失一段包含FHIT基因的DNA。对86例经显微切割的存档宫颈大环状电切术活检样本进行了评估，这些样本包括宫颈上皮内瘤变（CIN）1级（n = 27）、CIN3级（n = 30）和微浸润癌（n = 29）病例，检测HPV感染情况以及FHIT基因杂合性缺失（LOH）。使用荧光标记的基因内微卫星标记D3S1300和D3S4103，通过聚合酶链反应（PCR）检测FHIT基因LOH。使用Fragment Manager（商标）软件在半自动DNA测序仪上分析PCR产物，以确定等位基因缺失情况。使用通用引物和型特异性引物结合限制性内切酶消化，通过PCR确定病变的HPV状态。使用Epi-Info和SPSS-PC统计分析软件分析结果。对86例病例的苏木精和伊红染色切片分析了六种组织病理学特征，其中一些特征先前已被证明与微浸润癌有关。在CIN1病例中，36%发现FHIT基因LOH；CIN3病例中，52%发现FHIT基因LOH；微浸润病例中，73%发现FHIT基因LOH（p = 0.029）。在CIN3病例中，68%发现HPV 16 DNA；微浸润病例中，93%发现HPV 16 DNA（p < 0.001）。第二常见的HPV类型是HPV 31，仅在四个病变中发现，其中三个有FHIT基因LOH。当使用卡方检验评估FHIT基因LOH与HPV 16和31感染时，发现有统计学显著关系（p = 0.014）。发现FHIT基因LOH与所评估的组织病理学特征无关。FHIT基因LOH与致癌性HPV感染之间存在统计学显著关系，这表明在宫颈癌进展过程中病毒DNA整合与随后的基因缺失之间存在联系。FHIT基因异常可能被证明是早期子宫颈癌进展的优秀标志物。
