[Paraffin section thickness in immunohistochemical detection of p16 expression in cervical tissue samples]

Zhejiang Da Xue Xue Bao Yi Xue Ban. 2018 Feb 25;47(4):362-366. doi: 10.3785/j.issn.1008-9292.2018.08.06.
[Article in Chinese]

Abstract

Objective: To investigate paraffin section thickness in immunohistochemical detection of p16 expression in cervical tissue samples.

Methods: p16 immunohistochemical staining was performed in 150 cases of chronic cervicitis, 126 cases of low-grade squamous intraepithelial lesions(LSIL), 96 cases of high-grade squamous intraepithelial lesions (HSIL) and 78 cases of cervical cancer from January 2014 to March 2018 in Zhongshan Boai Hospital. The results of p16 protein expression in paraffin sections with thickness of 2, 3, 4, 5 and 6 μm were compared using Logistic regression analysis.

Results: With the increase of slice thickness, the staining of p16 protein in nucleus gradually increased. The thickness of cervical slices in chronic cervicitis and cervical cancer samples had no significant effect on the positive rate of p16 protein(χ2=7.817 and 1.332, both P>0.05), while the thickness of slices in LSIL and HSIL samples had significant effect on the positive rate of p16 protein (χ2=17.688 and 10.182, P<0.05 or P<0.01). The stable and reliable results were obtained when the slices were between 3 and 5 μm thick.

Conclusions: Paraffin sections with thickness of 3.0-5.0 μm are recommended for immnohistochemical staining of p16 protein in cervical tissue samples.

目的: 探讨免疫组织化学法检测子宫颈组织中p16蛋白表达时的石蜡切片厚度。

方法: 收集2014年1月至2018年3月中山市博爱医院病理检查结果为慢性宫颈炎(150例)、低级别鳞状上皮内病变(LSIL,126例)、高级别鳞状上皮内病变(HSIL,96例)及宫颈癌(78例)患者的子宫颈组织标本进行p16免疫组织化学染色,并采用条件Logistic回归分析石蜡切片厚度为2.0、3.0、4.0、5.0、6.0 μm的标本p16蛋白表达的差异。

结果: 随着切片厚度增加,病变部位所在细胞核p16蛋白染色逐渐加深。慢性宫颈炎和宫颈癌患者标本不同切片厚度的p16蛋白阳性率差异无统计学意义( χ 2=7.817和1.332,均 P>0.05);而LSIL和HSIL患者标本不同切片厚度的p16蛋白阳性率差异有统计学意义( χ 2=17.688和10.182, P < 0.05或 P < 0.01),当标本切片厚度为3.0~5.0 μm时可获得较为稳定而可靠的检测结果。

结论: 免疫组织化学法行子宫颈组织p16蛋白检测推荐的标本切片厚度为3.0~5.0 μm。

MeSH terms

  • Biomarkers, Tumor* / genetics
  • Biomarkers, Tumor* / metabolism
  • Cyclin-Dependent Kinase Inhibitor p16* / genetics
  • Cyclin-Dependent Kinase Inhibitor p16* / metabolism
  • Female
  • Histocytological Preparation Techniques* / standards
  • Humans
  • Immunohistochemistry
  • Paraffin*
  • Squamous Intraepithelial Lesions of the Cervix
  • Uterine Cervical Dysplasia*
  • Uterine Cervical Neoplasms* / physiopathology

Substances

  • Biomarkers, Tumor
  • Cyclin-Dependent Kinase Inhibitor p16
  • Paraffin

Grants and funding

广东省医学科研基金(A2017321);中山市社会公益科技研究专项资金(2018B1035);中山市卫生和计划生育局医学科研立项课题(2014J128);中山市卫生和计划生育局医学科研课题(2017J111)