中心体相关蛋白CDK5RAP2单克隆抗体的制备及其在乳腺癌中的表达  

Evaluation of CDK5RAP2 expression in breast cancer by preparing its monoclonal antibody

作　　者：张霖[1] 张彤雯[1] 杨毅[1] 李萌辉[1] 罗艺[1] 魏熙胤[1] 牛昀[1] 牛瑞芳[1] 齐众[2] 

机构地区：[1]天津医科大学附属肿瘤医院中心实验室国家教育部乳腺癌防治重点实验室,天津300060 [2]香港科技大学

出　　处：《中华乳腺病杂志（电子版）》2008年第5期31-34,共4页Chinese Journal of Breast Disease(Electronic Edition)

摘　　要：目的通过制备中心体相关蛋白CDK5RAP2的单克隆抗体,检测CDK5RAP2在正常乳腺及乳腺癌中的表达差异,为进一步研究CDK5RAP2在乳腺癌发生、发展中的作用提供依据。方法采用传统的杂交瘤方法制备CDK5RAP2的单克隆抗体,通过敲除CDK5RAP2的细胞模型进行免疫荧光检测,验证其特异性。选取正常乳腺、导管内癌和浸润性导管癌3组标本各30例,用制备的单克隆抗体进行免疫组化检测。结果得到一株CDK5RAP2单克隆抗体(5F6),亚型为IgG1,用免疫荧光检测确证其特异性。免疫组化检测CDK5RAP2的表达显示浸润性导管癌和导管内癌之间无明显的差别,但是两组均明显高于正常乳腺组(P<0.001)。结论乳腺正常组织不表达或低表达CDK5RAP2,而乳腺癌组织中该蛋白的表达率明显增高,提示CDK5RAP2可能成为乳腺癌诊治的新型标志物。Objective To evaluate the difference of CDKSRAP2 expression between normal breast tissue（N） and malignant breast cancer by its monoclonal antibody. Methods Monoclonal antibody of CDKSRAP2 was prepared by means of hybridoma technique and its specificity was tested by immunofluorescence on the cell knockout CDKSRAP2. Three groups of samples： normal breast tissue, ductal carcinoma in situ （DCIS） and infiltrating ductal carcinoma （IDC） （30 cases in each group） were selected. Immunohistochemistry（ IHC ） was performed on paraffin blocks by the CDKSRAP2 McAb to determine the expression patterns of CDKSRAP2 in breast cancer and normal breast tissue respectively. Results One antibody against CDKSRAP2 （5F6） with lgG1 subtype was obtained. IHC staining indicated that the CDKSRAP2 expression in both IDC and DCIS cases was higher than that in normal group （P 〈 0. 001 ）, but there was no difference between IDC and DCIS groups. Conclusion CDKSRAP2 is not expressed or lowly expressed in normal breast tissue. The CDKSRAP2 expression is increasingly enhanced in breast cancer tissues to lead to breast cancer occurrence and progressing. Therefore CDKSRAP2 may be a potential indicator of breast cancer.

关 键 词：CDKSRAP2 乳腺癌 单克隆抗体 

分 类 号：R737.9[医药卫生—肿瘤]