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中华乳腺病杂志(电子版) ›› 2008, Vol. 02 ›› Issue (05) : 31 -34. doi: 10.3877/cma.j.issn.1674-0807.2008.05.008

实验研究

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
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   
  1. 1.300060 天津,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
    2.300060 天津,天津医科大学附属肿瘤医院香港科技大学
  • 收稿日期:2008-01-29 出版日期:2008-10-01
  • 通信作者: 牛瑞芳

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

Lin ZHANG1, Tong-wen ZHANG1, Yi YANG1, Meng-hui LI1, Yi LUO1, Xi-yin WEI1, Yun NIU1, Ri-fang NIU,1(), Zhong QI1   

  1. 1.Central Laboratory, Tianjin Medical University Cancer Institute & Hospital, Tianjin 300060, China
  • Received:2008-01-29 Published:2008-10-01
  • Corresponding author: Ri-fang NIU
引用本文:

张霖, 张彤雯, 杨毅, 李萌辉, 罗艺, 魏熙胤, 牛昀, 牛瑞芳, 齐众. 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[J/OL]. 中华乳腺病杂志(电子版), 2008, 02(05): 31-34.

Lin ZHANG, Tong-wen ZHANG, Yi YANG, Meng-hui LI, Yi LUO, Xi-yin WEI, Yun NIU, Ri-fang NIU, Zhong QI. Evaluation of CDK5RAP2 expression in breast cancer by preparing its monoclonal antibody[J/OL]. Chinese Journal of Breast Disease(Electronic Edition), 2008, 02(05): 31-34.

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 CDK5RAP2 expression between normal breast tissue(N) and malignant breast cancer by its monoclonal antibody.

Methods

Monoclonal antibody of CDK5RAP2 was prepared by means of hybridoma technique and its specificity was tested by immunofluorescence on the cell knockout CDK5RAP2. 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 CDK5RAP2 McAb to determine the expression patterns of CDK5RAP2 in breast cancer and normal breast tissue respectively.

Results

One antibody against CDK5RAP2 (5F6) with IgG1 subtype was obtained. IHC staining indicated that the CDK5RAP2 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

CDK5RAP2 is not expressed or lowly expressed in normal breast tissue. The CDK5RAP2 expression is increasingly enhanced in breast cancer tissues to lead to breast cancer occurrence and progressing. Therefore CDK5RAP2 may be a potential indicator of breast cancer.

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