Preparation of a novel monoclonal antibody specific to DARPP-32.
DARPP-32 (dopamine and cAMP-regulated phosphoprotein) is the bifunctional protein that can act as a PP1 inhibitor or a PKA inhibitor. Here we have for the first time generated a monoclonal antibody against DARPP-32. Enzyme-linked immunosorbent assay (ELISA) and Western immunoblotting were used to screen the hybridomas. As a result, one MAb named Y8 (IgG1) was characterized, which bound to the native DARPP-32 protein. Then the MAb was used to detect the expression of DARPP-32 in gastric cancer cells and tissues. DARPP-32 was found to be downregulated in multidrug-resistant gastric cancer cells compared with drug-sensitive cells. DARPP-32 was also found to be upregulated in poorly differentiated gastric cancer tissues compared with well-differentiated and moderately differentiated gastric tissues. The results showed that DARPP-32 might play an important role in carcinogenesis and multidrug resistance of gastric cancer.
Yang
Jianjun
J
Department of Digestive Surgery, Xijing Hospital of Digestive Diseases, Fourth Military Medical University of China, Xi'an, Shaanxi Province, China.
Li
Mengbin
M
Hong
Liu
L
Zhang
Hongwei
H
Sun
Li
L
Wang
Weizhong
W
D000230:Adenocarcinoma / Q000175:diagnosis / Q000188:drug therapy / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification*; D000918:Antibody Specificity; D000970:Antineoplastic Agents / Q000494:pharmacology; D051937:Dopamine and cAMP-Regulated Phosphoprotein 32 / Q000235:genetics / Q000276:immunology*; D019008:Drug Resistance, Neoplasm; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D006825:Hybridomas; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D009101:Multiple Myeloma / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D013274:Stomach Neoplasms / Q000175:diagnosis / Q000188:drug therapy / Q000276:immunology*; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D051937:Dopamine and cAMP-Regulated Phosphoprotein 32; D011994:Recombinant Proteins