p16INK4A as a marker for cervical dyskaryosis:: CIN and cGIN in cervical biopsies and ThinPrep™ smears
p16INK4A as a marker for cervical dyskaryosis:: CIN and cGIN in cervical biopsies and ThinPrep™ smears
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DOI：
10.1136/jcp.56.1.56
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发表时间：
2003-01-01
影响因子：
3.4
通讯作者：
O'Leary, JJ
中科院分区：
文献类型：
--
作者：
Murphy, N;Ring, M;O'Leary, JJ
目的：通过组织切片和ThinPrep（TM）涂片检测p16（INK4A）作为宫颈发育不良鳞状细胞和腺体细胞生物标志物的潜力。方法：对22例宫颈正常组织、5例宫颈腺上皮内瘤变（cGIN）、38例宫颈上皮内瘤变I型（CIN1）、33例CIN2、46例CIN3和10例浸润性癌（8例鳞状癌和2例腺癌）进行p16（INK4A）表达的免疫细胞化学分析。所有样品均采用福尔马林固定，石蜡包埋，揭抗原后采用小鼠单克隆抗p16（INK4A）抗体进行免疫组化分析。采用0 - 3评分系统评估染色强度。此外，我们还检测了p16（INK4A）在12个正常ThinPrep涂片、1个cGIN涂片和20个轻、中、重度核异常涂片中的表达情况。人乳头瘤病毒（HPV）检测采用改进的SYBR绿色检测系统。采用荧光聚合酶链反应（PCR）和液相PCR进行HPV特异性分型。结果：正常宫颈组织均无p16（INK4A）免疫反应性。除1例CIN3病例外，本研究中所有病例的鳞状细胞和腺状细胞p16（INK4A）表达均呈阳性。在CIN1病例中，p16（INK4A）主要在细胞核表达，在CIN2、CIN3、cGIN和侵袭性病例中，p16（INK4A）在细胞核和细胞质中均有表达。所有HPV阳性的病例都表达p16（INK4A）蛋白，尽管并非所有p16阳性的病例都是HPV阳性。一般来说，在HPV阴性或含有低风险HPV型的病例中，p16（INK4A）染色强度较低。结论：这种过表达模式表明p16（INK4A）可能作为宫颈鳞状和腺状肿瘤病变的诊断标志物。此外，该技术可用于在ThinPrep涂片中识别单个核异常细胞。因此，p16（INK4A）是宫颈核发育不良的有用标志物。
Aim: To examine the potential of p16(INK4A) as a biomarker for dysplastic squamous and glandular cells of the cervix in tissue sections and ThinPrep(TM) smears.Methods: Immunocytochemical analysis of p16(INK4A) expression was performed on 22 normal cervical tissue samples, five cervical glandular intraepithelial neoplasia (cGIN), 38 cervical intraepithelial neoplasia I (CIN1), 33 CIN2, 46 CIN3, and 10 invasive cancer cases (eight squamous and two adenocarcinomas). All samples were formalin fixed and paraffin wax embedded, and immunohistochemical analysis was carried out using a mouse monoclonal anti-p16(INK4A) antibody after antigen unmasking. The staining intensity was assessed using a 0 to 3 scoring system. In addition, the expression status of p16(INK4A) was examined in 12 normal ThinPrep smears, one smear exhibiting cGIN, and a total of 20 smears exhibiting mild, moderate, and severe dyskaryosis. Human papillomavirus (HPV) detection was carried out using a modified SYBR green assay system. Fluorogenic polymerase chain reaction (PCR) and solution phase PCR were used for specific HPV typing.Results: p16(INK4A) immunoreactivity was absent in all normal cervical tissues examined. Dysplastic squamous and glandular cells were positive for p16(INK4A) expression in all cases included in this study, except for one CIN3 case. p16(INK4A) expression was mainly nuclear in CIN1 cases, and both nuclear and cytoplasmic in CIN2, CIN3, cGIN, and invasive cases. All cases positive for HPV expressed the p16(INK4A) protein, although not all cases found positive for p 16 were HPV positive. In general, the p16(INK4A) staining intensity was lower in cases negative for HPV or those containing a low risk HPV type.Conclusion: This pattern of overexpression demonstrates the potential use of p16(INK4A) as a diagnostic marker for cervical squamous and also glandular neoplastic lesions. In addition, the technique can be used to identify individual dyskaryotic cells in ThinPrep smears. Thus, p16(INK4A) is a useful marker of cervical dyskaryosis.