抗人BORIS单克隆抗体的制备及在乳腺疾病和正常乳腺组织中的分布

目的制备抗BORIS的单克隆抗体(mAb),并在蛋白水平上研究BORIS抗原在乳腺癌、良性乳腺病及正常乳腺组织(癌旁远端组织)上的分布特点。方法以人BORIS重组蛋白免疫BALB/c小鼠,应用B淋巴细胞杂交瘤技术建立分泌抗BORISmAb的杂交瘤细胞株。用免疫组化染色法检测BORIS抗原在睾丸组织、乳腺癌、乳腺良性病和正常乳腺组织(癌旁远端组织)中的表达。结果获得4株稳定分泌抗BORISmAb的杂交瘤细胞株。其中抗体编号为FMU-BORIS13(IgG1)可用于免疫组化的染色。结果显示,BORIS抗原在正常睾丸的精原细胞和精母细胞的细胞核内表达,符合CT抗原的表达特点。组织芯片的免疫组化染色结果显示其在乳腺癌组织中呈高水平的表达,阳性率为85%(94/110),在良性乳腺病组织中其表达的阳性率为95%(21/22)。另外,在6例乳腺癌癌旁远端近似于正常的乳腺组织中也检测到BORIS抗原的表达。BORIS抗原在乳腺癌组织中的表达远高于其在良性乳腺病和正常的乳腺组织(癌旁远端组织)中的表达。结论BORIS的高表达可能与乳腺癌的发生发展有一定的关系。

Preparation of the monoclonal antibodies against human BORIS and the expression pattern of BORIS in normal and diseased human mammary tissues

AIM: To prepare monoclonal antibodies(mAb) to BORIS antigen and analyze its expression pattern in breast cancer, benign breast disease and normal breast tissues. METHODS: Female BALB/c mice were immunized with recombination human BORIS protein. Hybridoma cell lines were established by hybridoma technique. BORIS antigen was detected by immunohistochemical (ICH) staining. RESULTS: Four clones of hybridomas stably secereting mAb against human BORIS were obtained. mAb FMMU-BORIS13 was selected to be employed in ICH. Positive staining was localized in the nuclei of the spermatocytes and spermatogoniums, which was consistent with the typical expression pattern of CT antigen. BORIS antigen was detected in 85%(94/110) of breast cancer, 95%(21/22) of benign breast diseases and 6 normal breast tissues(surrouding tumor free breast tissue). CONCLUSION: Four clones of anti-BORIS mAb have been successfully prepared. The expression of BORIS antigen is much stronger in breast cancer than that in benign breast disease and normal breast tissues, which indicate that BORIS may be involved in the pathogenesis of the breast cancer.