Negri G, Moretto G, Menia E, Vittadello F, Kasal A, Mian C, Egarter-Vigl E
Department of Pathology, Central Hospital Bolzano, Bolzano, Italy.
J Clin Pathol. 2006 Aug;59(8):827-30. doi: 10.1136/jcp.2005.030726. Epub 2006 Feb 7.
To evaluate the feasibility and value of a modified Papanicolaou counterstain for p16(INK4a) immunostaining in liquid-based cervicovaginal samples.
Immunocytochemical analyses were carried out with p16(INK4a) and modified Papanicolaou counterstain on 81 liquid-based samples, including 23 of within normal limits (WNL), 6 of low-grade squamous intraepithelial lesion (LSIL), 20 of high-grade squamous intraepithelial lesion (HSIL), 16 of atypical squamous cells of undetermined significance (ASC-US) and 16 of atypical squamous cells, high-grade lesion cannot be excluded (ASC-H). Results were compared with histological or cytological follow-up. For comparison, samples from 29 more cases (10 of LSIL, 10 of ASC-H and 9 of HSIL) were immunostained with p16(INK4a) and conventionally counterstained with haematoxylin. The intensity of immunostaining in cases of squamous intraepithelial lesion (SIL) was assessed using a 0-3 scoring system. Interobserver agreement was calculated by kappa statistics.
Expression of p16(INK4a) was detected in 3 of 23 cases of WNL, 4 of 6 cases of LSIL, all cases of HSIL, 5 of 16 cases of ASC-US and 13 of 16 cases of ASC-H. Excluding two cases with no residual dysplastic cells in the immunocytochemistry, all cases of cervical intraepithelial neoplasia (CIN)2 or CIN3 at follow-up expressed p16(INK4a) and none of the p16(INK4a)-negative cases showed a high-grade lesion at follow-up. No evident differences in pattern or intensity of p16(INK4a) expression were observed between the specimens of the study and control groups. Interobserver agreement was significantly better in the study group than in the group with conventional immunostaining (combined kappa 0.773 v 0.549; p<0.05), and still better, albeit statistically not significant, than with conventional immunostaining and cervical smear test together (combined kappa 0.773 v 0.642).
Immunocytochemistry with p16(INK4a) and modified Papanicolaou counterstain may add to the cervicovaginal cytology the full potentiality of p16(INK4a) without the need of a further slide and the risk of loss of dysplastic cells, yet maintaining the typical morphological features of the smear test.
评估改良巴氏复染法用于液基宫颈阴道样本中p16（INK4a）免疫染色的可行性和价值。
对81份液基样本进行p16（INK4a）免疫细胞化学分析及改良巴氏复染，包括23份正常范围（WNL）样本、6份低级别鳞状上皮内病变（LSIL）样本、20份高级别鳞状上皮内病变（HSIL）样本、16份意义不明确的非典型鳞状细胞（ASC-US）样本和16份不能排除高级别病变的非典型鳞状细胞（ASC-H）样本。将结果与组织学或细胞学随访结果进行比较。为作对照，对另外29例样本（10例LSIL、10例ASC-H和9例HSIL）进行p16（INK4a）免疫染色并用苏木精进行传统复染。采用0-3评分系统评估鳞状上皮内病变（SIL）病例的免疫染色强度。通过kappa统计量计算观察者间一致性。
在23例WNL样本中有3例检测到p16（INK4a）表达，6例LSIL样本中有4例，所有HSIL样本，16例ASC-US样本中有5例，16例ASC-H样本中有13例。排除免疫细胞化学中无残留发育异常细胞的2例样本后，随访中所有宫颈上皮内瘤变（CIN）2或CIN3病例均表达p16（INK4a），且p16（INK4a）阴性病例在随访中均未显示高级别病变。研究组和对照组样本在p16（INK4a）表达模式或强度上未观察到明显差异。研究组的观察者间一致性显著优于传统免疫染色组（合并kappa值0.773对0.549；p<0.05），且虽无统计学意义但仍优于传统免疫染色与宫颈涂片检查联合使用的情况（合并kappa值0.773对0.642）。
p16（INK4a）免疫细胞化学及改良巴氏复染法可在宫颈阴道细胞学检查中充分发挥p16（INK4a）的潜力，无需额外制片且无发育异常细胞丢失风险，同时保持涂片检查的典型形态特征。
