Murphy N, Ring M, Killalea A G, Uhlmann V, O'Donovan M, Mulcahy F, Turner M, McGuinness E, Griffin M, Martin C, Sheils O, O'Leary J J
Department of Pathology, Coombe Women's Hospital, Dublin 8, Ireland Trinity College Dublin, Dublin 2, Ireland.
J Clin Pathol. 2003 Jan;56(1):56-63. doi: 10.1136/jcp.56.1.56.
To examine the potential of p16(INK4A) as a biomarker for dysplastic squamous and glandular cells of the cervix in tissue sections and ThinPrep smears.
Immunocytochemical analysis of p16(INK4A) expression was performed on 22 normal cervical tissue samples, five cervical glandular intraepithelial neoplasia (cGIN), 38 cervical intraepithelial neoplasia 1 (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.
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 p16(INK4A) 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.
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.
检测p16（INK4A）作为子宫颈发育异常的鳞状和腺细胞在组织切片及ThinPrep涂片上生物标志物的潜力。
对22例正常宫颈组织样本、5例宫颈腺上皮内瘤变（cGIN）、38例宫颈上皮内瘤变1级（CIN1）、33例CIN2、46例CIN3以及10例浸润癌病例（8例鳞状细胞癌和2例腺癌）进行p16（INK4A）表达的免疫细胞化学分析。所有样本均用福尔马林固定并石蜡包埋，抗原修复后使用小鼠单克隆抗p16（INK4A）抗体进行免疫组织化学分析。染色强度采用0至3分评分系统进行评估。此外，在12例正常ThinPrep涂片、1例显示cGIN的涂片以及总共20例显示轻度、中度和重度核异质的涂片中检测p16（INK4A）的表达状态。使用改良的SYBR绿检测系统进行人乳头瘤病毒（HPV）检测。荧光定量聚合酶链反应（PCR）和液相PCR用于特定HPV分型。
在所检测的所有正常宫颈组织中，p16（INK4A）免疫反应性均缺失。除1例CIN3病例外，本研究纳入的所有病例中发育异常的鳞状和腺细胞p16（INK4A）表达均为阳性。在CIN1病例中，p16（INK4A）表达主要位于细胞核，而在CIN2、CIN3、cGIN和浸润性病例中，p16（INK4A）表达同时位于细胞核和细胞质。所有HPV阳性病例均表达p16（INK4A）蛋白，尽管并非所有p16（INK4A）阳性病例均为HPV阳性。一般来说，HPV阴性病例或含有低风险HPV类型的病例中，p16（INK4A）染色强度较低。
这种过表达模式表明p16（INK4A）有可能作为子宫颈鳞状和腺性肿瘤病变的诊断标志物。此外，该技术可用于识别ThinPrep涂片中的单个核异质细胞。因此，p16（INK4A）是子宫颈核异质的有用标志物。
