聚羟基丙烯酸和4%中性缓冲甲醛对苏木精伊红染色及荧光原位杂交检测乳腺癌HER-2基因影响的比较

doi:10.3969/j.issn.1007-3969.2016.2.001

中国癌症杂志 ›› 2016, Vol. 26 ›› Issue (2): 121-127.doi: 10.3969/j.issn.1007-3969.2016.2.001

聚羟基丙烯酸和4%中性缓冲甲醛对苏木精伊红染色及荧光原位杂交检测乳腺癌HER-2基因影响的比较

陈志强，王　莹，米贤军，陈　昂，钟守军，黄华勇，邓文同，刘超凡，徐秀梅，代新珍

南方医科大学附属中山博爱医院病理科，广东中山528400

出版日期:2016-02-29 发布日期:2016-06-01
通信作者: 陈志强　E-mail：765228687@qq.com
基金资助:
广东省医学科研基金立项课题(A2011746)；中山市科技计划项目重大专项(20113A004)；中山市局立项课题(2014J128)。

Comparison of the role that poly hydroxy acrylic acid plays in the detection of HER-2 gene in breast cancer by hematoxylin and eosin staining and fluorescence in situ hybridization with that of 4% neutral buffered formaldehyde

CHEN Zhiqiang, WANG Ying, MI Xianjun, CHEN Ang, ZHONG Shoujun, HUANG Huayong, DENG Wentong LIU Chaofan, XU Xiumei, DAI Xinzhen

Department of Pathology, Zhongshan Boai Hospital Affiliated to Southern Medical University, Zhongshan 528400, Guangdong Province, China

Published:2016-02-29 Online:2016-06-01
Contact: CHEN Zhiqiang E-mail: 765228687@qq.com

摘要/Abstract

摘要： 背景与目的：适当的组织固定、透明脱蜡是苏木精伊红(hematoxylin eosin，HE)染色及荧光原位杂交(fluorescence in situ hybridization，FISH)检测乳腺癌HER-2基因一个重要的步骤。该研究旨在对比环保固定液聚羟基丙烯酸和4%中性缓冲甲醛固定制作的组织切片HE染色优良率，荧光原位杂交检测两者固定制作的乳腺癌组织切片的HER-2基因扩增阳性率，探讨其替代传统固定液4%中性缓冲甲醛的可行性。方法：收集中山市博爱医院2011年3月—2015年1月门诊及住院部送检的乳腺癌69例、乳腺纤维腺瘤41例、子宫平滑肌瘤40例、宫颈组织25例、胎盘组织25例，各取材2块，每例标本的2块组织用抽签法随机分2组，一组采用4%中性缓冲甲醛固定制作HE切片200张，另一组采用聚羟基丙烯酸固定制作HE切片200张。依据切片染色情况判定切片等级，采用SPSS 19.0比较两者HE染色优良率；两组乳腺浸润性导管癌组织再各制作切片69张，采用SPSS 19.0比较FISH技术检测的HER-2基因扩增率。结果：① 聚羟基丙烯酸、4%中性缓冲甲醛固定组组织切片HE染色优质片分别为155张、166张，优良片41张、33张，中等片3张、1张，差片1张、0张，染色优良率分别为98.0%、99.5%，两组间优良率差异无统计学意义(χ²=1.33，P=0.25)。② 聚羟基丙烯酸、4%中性缓冲甲醛固定组组织切片FISH阳性率分别为26.09%、23.19%，两组间阳性率差异无统计学意义(χ²=0.50，P＞0.05)。结论：聚羟基丙烯酸固定制作的组织切片HE染色效果及荧光原位杂交检测乳腺癌HER-2基因扩增效果跟传统固定液4%中性缓冲甲醛相比差异无统计学意义，符合环保要求，有推广使用的可能。

关键词: 聚羟基丙烯酸, 甲醛, 组织切片, 苏木精伊红染色, 乳腺癌, HER-2基因, 荧光原位杂交

Abstract: Background and purpose: Adequate tissue fixation, transparent dewaxing is an important step of hematoxylin eosin (HE) staining and fluorescence in situ hybridization (FISH) in detection of breast cancer HER-2 gene. The purpose of this study was to make a comparison between poly hydroxyl acrylic acid which is an environmentally friendly fixation liquid and 4% neutral buffered formaldehyde in tissue fixation for HE staining and FISH to detect the HER-2 gene in the breast cancer tissue sections. The study aimed to evaluate the feasibility of replacing 4% buffered formaldehyde, a traditional fixation liquid, with the poly hydroxyl acrylic acid, an environmentally friendly fixation fluid. Methods: This project was performed on tissue samples collected from 69 cases of breast cancer, 41 cases of breast fibroadenoma, 40 cases of uterine leiomyoma, 25 cases of cervical tissue, 25 cases of placenta obtained from the outpatient and inpatient departments of Zhongshan Boai Hospital from Mar. 2011 to Jan. 2015, from each of which two samples were drawn and two blocks of each specimen were divided into two groups randomly. Then one group was fixed with 4% neutral buffered formaldehyde and made into 200 sections by HE while the other group was fixed with poly hydroxyl acrylic acid and made into another 200 sections. The slice level of the two groups was determined by the staining condition of the sections, and SPSS 19.0 was employed to compare the excellent and good rate of HE staining. Additional 69 sections were produced with two groups of breast invasive ductal carcinoma tissues, and SPSS 19.0 was used to detect the amplification of HER-2 gene by FISH. Results: First, the number of best-quality slices stained with HE fixed separately by poly hydroxyl acrylic acid and 4% neutral buffered formaldehyde was 155 and 166, respectively. The number of excellent pieces was 41 and 33, respectively, while the number of mediocre pieces was 3 and 1 with bad pieces being 1 and 0, respectively. The excellent and good rates of HE staining were 98% and 99.5%, respectively. There was no significant difference between the two groups (χ²=1.33, P>0.05).Second, the positive rates of the tissue slices by FISH fixed separately by poly hydroxyl acrylic acid and 4% neutral buffered were 26.09% and 23.19%, respectively. There was no significant difference between the two groups (χ²=0.50, P>0.05).Conclusion: The results obtained with HE staining and FISH using poly hydroxyl acrylic acid as a fixation liquid are not significantly different from those using 10% neutral buffered formaldehyde. Therefore, poly hydroxyl acrylic acid meets the requirements of environmental protection, and thus has the potential to be promoted and widely used.

Key words: Poly hydroxyl acrylic acid, Formaldehyde, Tissue section, Hematoxylin eosin staining, Breast cancer, HER-2 gene, Fluorescence in situ hybridization

陈志强,王　莹,米贤军,等. 聚羟基丙烯酸和4%中性缓冲甲醛对苏木精伊红染色及荧光原位杂交检测乳腺癌HER-2基因影响的比较[J]. 中国癌症杂志, 2016, 26(2): 121-127.

CHEN Zhiqiang, WANG Ying, MI Xianjun, et al. Comparison of the role that poly hydroxy acrylic acid plays in the detection of HER-2 gene in breast cancer by hematoxylin and eosin staining and fluorescence in situ hybridization with that of 4% neutral buffered formaldehyde[J]. China Oncology, 2016, 26(2): 121-127.

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聚羟基丙烯酸和4%中性缓冲甲醛对苏木精伊红染色及荧光原位杂交检测乳腺癌HER-2基因影响的比较

Comparison of the role that poly hydroxy acrylic acid plays in the detection of HER-2 gene in breast cancer by hematoxylin and eosin staining and fluorescence in situ hybridization with that of 4% neutral buffered formaldehyde

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