Establishment of an ELISA to assess the autoantibody against hyaluronan and proteoglycan link protein 1 (HAPLN1) in canine sera for detection of canine mammary complex carcinoma

• Main Authors: Huan-Ming Chang, 張桓鳴
• Other Authors: 劉浩屏
• Format: Others
• Language: zh-TW
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/27d45h

碩士 === 國立中興大學 === 獸醫學系暨研究所 === 106 === Canine mammary tumors (CMT) are the most common neoplasms in sexually intact female dogs. About 40-50% of CMT cases are malignant, most of which are of epithelial origin and are classified into several types. Among these malignant tumor types, complex carcinoma is the most prevalent, whereas there are no reliable biomarkers for detection of this malignance. Our lab previously has identified a number of proteins significantly overexpressed in complex carcinoma compared with normal mammary tissues, one of which is Hyaluronan And Proteoglycan Link Protein 1 (HAPLN1) a secretory protein involved in extracellular matrix assembly. Overexpression of HAPLN1 in human cancers is associated with cancer progression and bad outcome. As a tumor-associated protein, HAPLN1 is shown to induce auto-antibodies (AAbs) accessible in human sera. The present study aims at establishing an ELISA to assess HAPLN1 AAbs in canine sera and evaluate the effectiveness of using HAPLN1 AAbs in detection of canine mammary carcinoma. In the ELISA, the HAPLN1 recombinant protein expressed in E. coli BL21(DE3) was used as the antigen for detection of seral HAPLN1 AAbs. Serum samples from CMT dogs (n = 69) and female healthy donors (n = 26) were collected and subjected to the ELISA. Among all CMT groups, the level of seral HAPLN1 AAbs was significantly higher in the dogs with complex carcinoma (n = 23) compared with healthy donors (P = 0.0185). In the aspect of clinical stage, the level of HAPLN1 AAbs was significantly higher in dogs with early-stage carcinoma (n = 36) compared with healthy donors (P = 0.0192), whereas there was no significance between stages. When stratifying all the groups by age of 8 yr and over, it was significant in comparisons of the level of HAPLN1 AAbs between complex carcinoma (n = 21) and healthy donors (n = 7), early-stage carcinoma (n = 31) and healthy donors, and early-stage complex carcinoma (n = 16) and healthy donors (P = 0.0035, P = 0.0019, and P = 0.0040, respectively). In sum, this study demonstrates for the first time that HAPLN1 AAbs are accessible in sera of dogs with CMTs and hold potential to be used as serological tools for early detection of CMTs, particularly complex carcinoma.