Please wait a minute...
J Zhejiang Univ (Med Sci)  2018, Vol. 47 Issue (4): 362-366    DOI: 10.3785/j.issn.1008-9292.2018.08.06
Paraffin section thickness in immunohistochemical detection of p16 expression in cervical tissue samples
CHEN Zhiqiang(),MI Xianjun,CHEN Ang*(),DUAN Lifeng,DAI Xinzhen,DENG Wentong
Department of Pathology, Zhongshan Boai Hospital Affiliated to Southern Medical University, Zhongshan 528400, Guangdong Province, China
Download: HTML( 2 )   PDF(1031KB)
Export: BibTeX | EndNote (RIS)      


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

Key wordsUterine cervical neoplasms/pathology      Uterine cervical neoplasms/diagnosis      Carcinoma, squamous cell/pathology      Immunohistochemistry      Genes, p16      Paraffin embedding      Microtomy     
Received: 12 April 2018      Published: 04 December 2018
CLC:  R365  
Corresponding Authors: CHEN Ang     E-mail:;
Cite this article:

CHEN Zhiqiang,MI Xianjun,CHEN Ang,DUAN Lifeng,DAI Xinzhen,DENG Wentong. Paraffin section thickness in immunohistochemical detection of p16 expression in cervical tissue samples. J Zhejiang Univ (Med Sci), 2018, 47(4): 362-366.

URL:     OR


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(χ2=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(χ2=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。

关键词: 宫颈肿瘤/病理学,  宫颈肿瘤/诊断,  癌, 鳞状细胞/病理学,  免疫组织化学,  基因, p16,  石蜡包埋,  显微切片术 
Fig 1 Expression of p16 protein in cervical slices of different thickness in patients with high-grade squamous intraepithelial lesions under microscope
切片厚度(μm) n 2.0 3.0 4.0 5.0 6.0 χ2 P
LSIL:低级别鳞状上皮内病变;HSIL:高级别鳞状上皮内病变.与2.0 μm比较,*P<0.05,* *P<0.01;与5.0 μm比较,#P<0.05,##P<0.01;与6.0 μm比较,P<0.05,△△P<0.01.
慢性宫颈炎 150 2(1.3) 8(5.3)# 10(6.7)*△ 8(5.3) 2(1.33) 7.817 >0.05
LSIL 126 22(17.5) 42(33.3)*##△△ 44(34.9)**##△△ 46(36.5)**△△ 20(15.9) 17.688 <0.01
HSIL 96 56(58.3) 78(81.3) 76(79.2) 76(79.2) 50(52.1) 10.182 <0.05
宫颈癌 78 70(89.7) 78(100.0) 78(100.0) 78(100.0) 68(87.2) 1.332 >0.05
Tab 1 Effect of slice thickness on the positive rate of p16 protein in cervical tissues of different lesions
[1]   魏丽惠 . HPV感染现状及在宫颈癌和癌前病变筛查中的意义[J]. 实用妇产科杂志, 2017, 33 (2): 81- 83
WEI Lihui . The status of HPV infection and its significance in screening cervical cancer and precancerous lesions[J]. Journal of Practical Obstetrics and Gynecology, 2017, 33 (2): 81- 83
[2]   GUSTINUCCI D , PASSAMONTI B , CESARINI E et al. Role of p16(INK4a) cytology testing as an adjunct to enhance the diagnostic specificity and accuracy in human papillomavirus-positive women within an organized cervical cancer screening program[J]. Acta Cytol, 2012, 56 (5): 506- 514
doi: 10.1159/000338979
[3]   郭艳敏, 周梅香 . p16、Ki-67在宫颈上皮内病变中的表达及意义[J]. 诊断病理学杂志, 2017, 24 (2): 105- 107
GUO Yanmin , ZHOU Meixiang . Expression of p16 and Ki-67 in cervical intraepithelial lesions and their significance[J]. Chinese Journal of Diagnostic Pathology, 2017, 24 (2): 105- 107
doi: 10.3969/j.issn.1007-8096.2017.02.006
[4]   中华医学会 . 临床技术操作规范病理学分册[M]. 北京: 人民军医出版社, 2012: 32- 33
Chinese Medical Association . Pathology of clinical practice[M]. Beijing: People's Military Medical Press, 2012: 32- 33
[5]   金蓉蓉, 马红伟, 陈天羽 et al. p16/Ki67双染检测在意义不明的不典型鳞状细胞分流诊断中的作用[J]. 中华病理学杂志, 2017, 46 (7): 481- 484
JIN Rongrong , MA Hongwei , CHEN Tianyu et al. Application of p16/Ki-67 immunocytochemistry in triage of patients with atypical squamous cells of undetermined significance[J]. Chinese Journal of Pathology, 2017, 46 (7): 481- 484
doi: 10.3760/cma.j.issn.0529-5807.2017.07.007
[6]   YOKCHOM R , LAIWEJPITHAYA S , MANEEPRAKORN W et al. Paper-based immunosensor with signal amplification by enzyme-labeled anti-p16INK4a multifunctionalized gold nanoparticles for cervical cancer screening[J]. Nanomedicine, 2018, 14 (3): 1051- 1058
doi: 10.1016/j.nano.2018.01.016
[7]   TSAKOGIANNIS D , MOSCHONAS G D , BELLA E et al. Association of p16(CDKN2A) polymorphisms with the development of HPV16-related precancerous lesions and cervical cancer in the Greek population[J]. J Med Virol, 2018, 90 (5): 965- 971
doi: 10.1002/jmv.v90.5
[8]   SARITHA V N , VEENA V S , JAGATHNATH K K M et al. Significance of DNA replication licensing proteins (MCM2, MCM5 and CDC6), p16 and p63 as markers of premalignant lesions of the uterine cervix:its usefulness to predict malignant potential[J]. Asian Pac J Cancer Prev, 2018, 19 (1): 141- 148
[9]   KANTHIYA K , KHUNNARONG J , TANGJITGAMOL S et al. Expression of the p16 and Ki67 in cervical squamous intraepithelial lesions and cancer[J]. Asian Pac J Cancer Prev, 2016, 17 (7): 3201- 3206
[10]   张谷, 周芳, 毛伟敏 et al. 乳腺癌石蜡切片厚度对荧光原位杂交检测HER2和TOP2A结果的影响[J]. 诊断病理学杂志, 2015, 44 (5): 334- 337
ZHANG Gu , ZHOU Fang , MAO Weimin et al. Effect of paraffin section thickness of breast cancer on the results of HER2 and TOP2A detected by fluorescence in situ hybridization[J]. Chinese Journal of Diagnostic Pathology, 2015, 44 (5): 334- 337
[11]   VARGA Z , TUBBS R R , WANG Z et al. Co-amplification of the HER2 gene and chromosome 17 centromere:a potential diagnostic pitfall in HER2 testing in breast cancer[J]. Breast Cancer Res Treat, 2012, 132 (3): 925- 935
doi: 10.1007/s10549-011-1642-8
[1] LU Zhanjun, HU Yangyang, LI Sisi, ZANG Lijuan, JIANG Weiliang, WU Jianjiong, WU Xiening, ZENG Yue, WANG Xingpeng. Anti-inflammatory effect of interleukin-35 in mice with colitis and its mechanism[J]. J Zhejiang Univ (Med Sci), 2018, 47(5): 499-506.
[2] CHEN Qian,LIU Lu,ZHANG Jingjing,HAN Sai,CUI Baoxia,ZHANG Youzhong,KONG Beihua. Clinical features and prognosis of cervical adenocarcinoma and adenosquamous carcinoma: an analysis of 237 cases[J]. J Zhejiang Univ (Med Sci), 2018, 47(4): 357-361.
[3] DI Chenhong,JIN Fan. Value of combined detection of claudin 4 and high-risk human papilloma virus in high-grade squamous intraepithelial lesion and cervix squamous cell carcinoma[J]. J Zhejiang Univ (Med Sci), 2018, 47(4): 344-350.
[4] HU Zheng,MA Ding. Precision screening and treatment of human papilloma virus related cervical cancer[J]. J Zhejiang Univ (Med Sci), 2018, 47(4): 338-343.
[5] YAO Guorong,FU Yunfeng,LI Yanli,ZHOU Caiyun,LV Weiguo. Expression and clinical significance of DNAJB11 in epithelial ovarian cancer[J]. J Zhejiang Univ (Med Sci), 2017, 46(2): 173-178.
[6] CAI Cheng,WANG Jianping,ZHONG Zhifeng,DAI Zhihui,WANG Qinghua,DONG Wuzhen,SHI Hongqi,LIU Qingwei,DU Jinlin. Hypoxia-inducible factor-1α and CD133 predicts pathological complete response and survival for locally advanced rectal cancer patients after neoadjuvant chemoradiotherapy[J]. J Zhejiang Univ (Med Sci), 2017, 46(1): 36-43.
[7] CHEN Xiaojing, XU Junfen, YE Jing, CHENG Xiaodong, XIE Xing, LYU Weiguo. Expression of miR-let-7e-3p in cervical intraepithelial neoplasm and cervix carcinoma and its clinical significance[J]. J Zhejiang Univ (Med Sci), 2016, 45(4): 342-348.
[8] ZHENG Jiangjiang, ZHU Yin, LI Changshui, LI Yinya, NIE Qianqian, ZHU Ziling, DENG Hong. Expression of CD10 in tumor-associated fibroblast of cancerized or recurrent colorectal adenomas[J]. J Zhejiang Univ (Med Sci), 2016, 45(4): 335-341.
[9] LAI Maode. Gastroenteropancreatic neuroendocrine neoplasms: concepts and related issues[J]. J Zhejiang Univ (Med Sci), 2016, 45(1): 5-9,23.
[10] YANG Yan, LI Yu-mei, ZHANG Na, LI Wan-yun, OU Yu-rong, WANG Rui, ZHAO Fu-you, WU Qiong. Expression of gap junction protein connexin 26 in human hepatocellular carcinoma and its significance[J]. J Zhejiang Univ (Med Sci), 2015, 44(5): 517-524.
[11] LIN Kai-qing, ZHU Li-bo, ZHANG Xin-mei, LIN Jun. Role of mast cells in estrogen-mediated experimental endometriosis in rats[J]. J Zhejiang Univ (Med Sci), 2015, 44(3): 269-277.
[12] LIU Xue-hong, ZHANG Yong, LIU Chuan-kang. Expression of neuron-specific enolase and synaptophysin in esophageal development of human embryos[J]. J Zhejiang Univ (Med Sci), 2015, 44(2): 184-188.
[13] OU Yu-rong,et al. Expression of secreted frizzled related protein 1,β-catenin and E-cadherin in colorectal carcinoma and its clinicopathological significances [J]. J Zhejiang Univ (Med Sci), 2014, 43(4): 397-405.
[14] CAO Pei-long,et al. Expressions of FOXC1 and MMP-7 in molecular subtypes of breast cancer and their association with clinicopathological characteristics [J]. J Zhejiang Univ (Med Sci), 2014, 43(4): 406-412.
[15] JIN Shu-qing,et al. Correlation of PBX2/ELF2 expression with prognosis of non-small cell lung cancer[J]. J Zhejiang Univ (Med Sci), 2014, 43(4): 413-419.