Pratt M Margaret, Sirajuddin Paul, Poirier Miriam C, Schiffman Mark, Glass Andrew G, Scott David R, Rush Brenda B, Olivero Ofelia A, Castle Philip E
Carcinogen-DNA Interactions Section, LCBG, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, USA.
Mutat Res. 2007 Nov 1;624(1-2):114-23. doi: 10.1016/j.mrfmmm.2007.04.008. Epub 2007 May 5.
Among women infected with carcinogenic human papillomavirus (HPV), there is a two- to five-fold increased risk of cervical precancer and cancer in women who smoke compared to those who do not smoke. Because tobacco smoke contains carcinogenic polycyclic aromatic hydrocarbons (PAHs), it was of interest to examine human cervical tissue for PAH-DNA adduct formation. Here, we measured PAH-DNA adduct formation in cervical biopsies collected in follow-up among women who tested positive for carcinogenic HPV at baseline. A semi-quantitative immunohistochemistry (IHC) method using antiserum elicited against DNA modified with r7,t8-dihydroxy-t-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BPDE) was used to measure nuclear PAH-DNA adduct formation. Cultured human cervical keratinocytes exposed to 0, 0.153, or 0.331microM BPDE showed dose-dependent increases in r7,t8,t9-trihydroxy-c-10-(N(2)deoxyguanosyl)-7,8,9,10-tetrahydro-benzo[a]pyrene (BPdG) adducts. For BPdG adduct analysis, paraffin-embedded keratinocytes were stained by IHC with analysis of nuclear color intensity by Automated Cellular Imaging System (ACIS) and, in parallel cultures, extracted DNA was assayed by quantitative BPDE-DNA chemiluminescence immunoassay (CIA). For paraffin-embedded samples from carcinogenic HPV-infected women, normal-appearing cervical squamous epithelium suitable for scoring was found in samples from 75 of the 114 individuals, including 29 cases of cervical precancer or cancer and 46 controls. With a lower limit of detection of 20 adducts/10(8) nucleotides, detectable PAH-DNA adduct values ranged from 25 to 191/10(8) nucleotides, with a median of 75/10(8) nucleotides. PAH-DNA adduct values above 150/10(8) nucleotides were found in eight samples, and in three samples adducts were non-detectable. There was no correlation between PAH-DNA adduct formation and either smoking or case status. Therefore, PAH-DNA adduct formation as measured by this methodology did not appear related to the increased risk of cervical precancer and cancer among carcinogenic HPV-infected smokers.
在感染致癌性人乳头瘤病毒（HPV）的女性中，与不吸烟的女性相比，吸烟女性发生宫颈上皮内瘤变和宫颈癌的风险增加了2至5倍。由于烟草烟雾中含有致癌性多环芳烃（PAH），因此研究人宫颈组织中PAH-DNA加合物的形成情况很有意义。在此，我们测量了在基线时致癌性HPV检测呈阳性的女性随访期间收集的宫颈活检组织中PAH-DNA加合物的形成情况。使用针对经r7,t8-二羟基-t-9,10-氧代-7,8,9,10-四氢苯并[a]芘（BPDE）修饰的DNA产生的抗血清的半定量免疫组织化学（IHC）方法来测量细胞核PAH-DNA加合物的形成。暴露于0、0.153或0.331微摩尔BPDE的培养人宫颈角质形成细胞显示出r7,t8,t9-三羟基-c-10-（N(2)-脱氧鸟苷基）-7,8,9,10-四氢苯并[a]芘（BPdG）加合物呈剂量依赖性增加。对于BPdG加合物分析，用IHC对石蜡包埋的角质形成细胞进行染色，并通过自动细胞成像系统（ACIS）分析细胞核颜色强度，并且在平行培养物中，通过定量BPDE-DNA化学发光免疫测定法（CIA）检测提取的DNA。对于来自致癌性HPV感染女性的石蜡包埋样本，在114名个体中的75名个体的样本中发现了适合评分的外观正常的宫颈鳞状上皮，包括29例宫颈上皮内瘤变或癌症病例和46名对照。检测下限为20个加合物/10(8)个核苷酸，可检测到的PAH-DNA加合物值范围为25至
