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The Anti-Proliferative Effect of a Newly-Produced Anti-PSCA-Peptide Antibody by Multiple Bioinformatics Tools, on Prostate Cancer Cells
Recent Patents on Anti-Cancer Drug Discovery ( IF 2.8 ) Pub Date : 2021-01-31 , DOI: 10.2174/1574892815999201110212411
Milad Chizari 1 , Sajad Fani-Kheshti 1 , Jaleh Taeb 1 , Mohammad M Farajollahi 1 , Monireh Mohsenzadegan 2
Affiliation  

Background: Prostate Stem Cell Antigen (PSCA) is a small cell surface protein, overexpressed in 90% of prostate cancers. Determination of epitopes that elicit an appropriate response to the antibody generation is vital for diagnostic and immunotherapeutic purposes for prostate cancer treatment. Presently, bioinformatics B-cell prediction tools can predict the location of epitopes, which is uncomplicated, faster, and more cost-effective than experimental methods.

Objective: We aimed to predict a novel linear peptide for Prostate Stem Cell Antigen (PSCA) protein in order to generate anti-PSCA-peptide (p) antibody and to investigate its effect on prostate cancer cells.

Methods: In the current study, a novel linear peptide for PSCA was predicted using in silico methods that utilize a set of linear B-cell epitope prediction tools. Polyclonal antibody (anti-PSCA-p antibody “Patent No. 99318”) against PSCA peptide was generated. The antibody reactivity was determined by the Enzyme-Linked Immunosorbent Assay (ELISA) and its specificity by immunocytochemistry (ICC), immunohistochemistry (IHC), and Western Blotting (WB) assays. The effect of the anti-PSCA-p antibody on PSCA-expressing prostate cancer cell line was assessed by Methylthiazolyldiphenyl- Tetrazolium bromide (MTT) assay.

Results: New peptide-fragment of PSCA sequence as “N-CVDDSQDYYVGKKN-C” (PSCA-p) was selected and synthesized. The anti-PSCA-p antibody against the PSCA-p showed immunoreactivity with PSCA-p specifically bound to PC-3 cells. Also, the anti-PSCA-p antibody strongly stained the prostate cancer tissues as compared to Benign Prostatic Hyperplasia (BPH) and normal tissues (P < 0.001). As the degree of malignancy increased, the staining intensity was also elevated in prostate cancer tissue (P < 0.001). Interestingly, the anti-PSCA-p antibody showed anti-proliferative effects on PC-3 cells (31%) with no growth inhibition effect on PSCA-negative cells.

Conclusion: In this study, we developed a new peptide sequence (PSCA-p) of PSCA. The PSCA-p targeting by anti-PSCA-p antibody inhibited the proliferation of prostate cancer cells, suggesting the potential of PSCA-p immunotherapy for future prostate cancer studies.



中文翻译:

通过多种生物信息学工具新产生的抗 PSCA 肽抗体对前列腺癌细胞的抗增殖作用

背景:前列腺干细胞抗原 (PSCA) 是一种小细胞表面蛋白,在 90% 的前列腺癌中过表达。确定引发对抗体生成的适当反应的表位对于前列腺癌治疗的诊断和免疫治疗目的至关重要。目前,生物信息学 B 细胞预测工具可以预测表位的位置,这比实验方法简单、快速且更具成本效益。

目的:我们旨在预测前列腺干细胞抗原 (PSCA) 蛋白的新型线性肽,以产生抗 PSCA 肽 (p) 抗体并研究其对前列腺癌细胞的影响。

方法:在目前的研究中,使用计算机方法预测 PSCA 的新型线性肽,该方法利用一组线性 B 细胞表位预测工具。产生了针对 PSCA 肽的多克隆抗体(抗 PSCA-p 抗体“专利号 99318”)。抗体反应性由酶联免疫吸附测定 (ELISA) 确定,其特异性由免疫细胞化学 (ICC)、免疫组织化学 (IHC) 和蛋白质印迹 (WB) 测定确定。抗PSCA-p抗体对表达PSCA的前列腺癌细胞系的作用通过甲基噻唑基二苯基-溴化四唑(MTT)测定法评估。

结果:选择并合成了PSCA序列的新肽片段“N-CVDDSQDYYVGKKN-C”(PSCA-p)。针对 PSCA-p 的抗 PSCA-p 抗体显示出与与 PC-3 细胞特异性结合的 PSCA-p 的免疫反应性。此外,与良性前列腺增生 (BPH) 和正常组织相比,抗 PSCA-p 抗体强烈染色前列腺癌组织 (P < 0.001)。随着恶性程度的增加,前列腺癌组织中的染色强度也升高(P < 0.001)。有趣的是,抗 PSCA-p 抗体对 PC-3 细胞(31%)显示出抗增殖作用,而对 PSCA 阴性细胞没有生长抑制作用。

结论:在本研究中,我们开发了 PSCA 的新肽序列 (PSCA-p)。抗 PSCA-p 抗体靶向 PSCA-p 抑制了前列腺癌细胞的增殖,表明 PSCA-p 免疫疗法在未来前列腺癌研究中具有潜力。

更新日期:2021-01-31
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