Preparation Of AKR1b10 Specific Antibody And Its Anti-breast Cancer Activity

Breast cancer has become one of the most common and serious malignant tumors among women in today's society,but its incidence is still increasing year by year.According to the current research on the expression of AKR1B10,AKR1B10 is highly expressed in gastrointestinal epithelial tissues,but it is low or not expressed in other tissues.However,it has been found that AKR1B10 is highly expressed in breast cancer,liver cancer,non-small cell lung cancer and other tumors.In recent years,many scholars have attached great importance to relevant research.Studies have shown that the up-regulation of AKR1B10 gene expression in breast cancer patients may be related to the involvement of AKR1B10 gene in lipid synthesis of ACCA(acetyl coenzyme A carboxylase alpha).The abnormal activity of some transcription factors can inhibit apoptosis,stimulate cell proliferation,and participate in the occurrence and development of tumors,and have a certain correlation with the clinical stage and prognosis of tumors.Objective:Based on breast cancer cells,we constructed endogenous BT-20 cells with high expression,MCF-7 cells with transcriptional AKR1B10 gene(MCF-7/AKR1B10 cells)and MCF-7 cells with transcriptional empty plasmid(MCF-7/p CDH cells).Then we studied the effects of AKR1B10 specific antibody,AKR1B10 inhibitor and Prunella vulgaris extract on the three groups of model cells,so as to explore the specific anti-AKR1B10.The body may provide a scientific basis for targeted treatment of breast cancer.Methods: Firstly,the recombinant AKR1B10 protein was purified based on the principle of Ni-NTA binding histidine.Purification of AKR1B10 Specific Antibody by Preparative Antigen Column.Then the following three aspects are discussed:(1)the effect of AKR1B10 specific antibody on BT-20 cells,MCF-7/AKR1B10 cells and MCF-7/p CDH cells.(2)The effects of AKR1B10 inhibitor on BT-20 cells,MCF-7/AKR1B10 cells and MCF-7/p CDH cells.(3)Effect of Prunella vulgaris extract on BT-20 cells,MCF-7/AKR1B10 cells and MCF-7/p CDH cells.Through these three aspects of research work and comprehensive research results,we will clarify the effect of AKR1B10 specific antibody on breast cancer cells after blocking AKR1B10 secretion.Results:(1)The concentration of recombinant AKR1B10 protein detected by BCA protein quantitative method was 1.32mg/ml.Coomassie brilliant blue staining confirmed that the purified recombinant AKR1B10 protein was of good purity and suitable for this experiment.(2)The concentration of AKR1B10 specific antibody detected by BCA protein quantitative method is 0.54mg/ml,and Coomassie brilliant blue staining confirms that the purity of AKR1B10 specific antibody is good,which is suitable for this experiment.(3)AKR1B10 specific antibody can inhibit the growth,proliferation and migration of breast cancer cells expressing AKR1B10,and can inhibit the growth,proliferation and migration of breast cancer cells expressing AKR1B10 more than AKR1B10 inhibitor and Prunella vulgaris extract.Conclusion: 1.AKR1B10 specific antibody can inhibit the growth,proliferation and migration of breast cancer cells expressing AKR1B10.2.AKR1B10 specific antibody may be used in the treatment of breast cancer.