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| Preparation of antibody against ANKRD22 and its expression in colorectal cancer |
| LIN Yun-hua*, LIU Jing-wen, YANG Sai-sai, LIU Xi-yong, WANG Hong-ping
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| *First People's Hospital of Pinghu,Pinghu,Zhejiang 314200,China |
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Abstract Objective To prepare the monoclonal antibody against ankyrin repeat domain 22（ANKRD22）and to investigate its expression in colorectal cancer tissues.Methods The recombinant human ANKRD22 was expressed through E. coli and pET-42a and then used to immunize Balb/c mice after purification. Anti-human ANKRD22 specific monoclonal antibodies were selected by Western blotting with 293T cell lysate highly expressing ANKRD22 as antigen. The expression of ANKRD22 in the tissue microarrays of 112 patients with colorectal cancer was detected by immunohistochemical staining.Results Four specific monoclonal antibodies against human ANKRD22 were screened out of 93 hybridoma cells,which reacted well with natural human ANKRD22. ANKRD22 was mainly distributed in the cytoplasm of colorectal cancer cells. In 112 cases of colorectal cancer,94 cases were detected positive for ANKRD22 expression,with the positive rate of 83.93%. The expression of ANKRD22 was statistically correlated with the expression of p53 and β-catenin（P<0.05）,but not with age,sex,location of tumors,AJCC stage,Dukes stage,degree of differentiation,lymph node metastasis and mismatch repair gene expression（P>0.05）.Conclusion The expression level of ANKRD22 was high in colorectal cancer. ANKRD22 might be involved in the carcinogenesis of colorectal epithelium and be a potential diagnostic marker.
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Received: 14 November 2018
Revised: 30 December 2018
Published: 04 March 2019
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