ARID1A is a subunit of the mammalian SWI/SNF complex, which is thought to regulate gene expression through restructuring chromatin structures. Its geneARID1Ais frequently mutated and ARID1A levels are lowered in several human cancers, especially gynecologic ones. A functional ARID1A loss may have prognostic or predictive value in terms of therapeutic strategies but has not been proposed based on a quantitative method. Hardly any literature is available on ARID1A levels in tumor samples. We developed an indirect enzyme-linked immunosorbent assay (ELISA) for ARID1A based on the current EMA and FDA criteria. We demonstrated that our ELISA provides the objective, accurate, and precise quantification of ARID1A concentrations in recombinant protein solutions, cell culture standards, and tissue lysates of tumors. A standard curve analysis yielded a ‘goodness of fit’ of R2= 0.99. Standards measured on several plates and days achieved an inter-assay accuracy of 90.26% and an inter-assay precision with a coefficient of variation of 4.53%. When tumor lysates were prepared and measured multiple times, our method had an inter-assay precision with a coefficient of variation of 11.78%. We believe that our suggested method ensures a high reproducibility and can be used for a high sample throughput to determine the ARID1A concentration in different tumor entities. The application of our ELISA on various tumor and control tissues will allow us to explore whether quantitative ARID1A measurements in tumor samples are of predictive value.
ARID1A是哺乳动物SWI/SNF复合物的亚基，该复合物被认为通过重构染色质结构来调控基因表达。其编码基因ARID1A在多种人类癌症（尤其是妇科肿瘤）中频繁发生突变且表达水平降低。ARID1A功能缺失可能对治疗策略具有预后或预测价值，但目前尚未建立基于定量方法的评估体系。关于肿瘤样本中ARID1A表达水平的文献极为有限。本研究依据现行EMA和FDA标准，开发了一种间接酶联免疫吸附测定法（ELISA）用于检测ARID1A。实验证明，该方法能够对重组蛋白溶液、细胞培养标准品及肿瘤组织裂解液中的ARID1A浓度进行客观、准确、精密的定量分析。标准曲线分析显示拟合优度R²=0.99。多板多日检测的标准品测定结果显示，批间准确度为90.26%，批间精密度变异系数为4.53%。对多次制备的肿瘤裂解液进行检测时，本方法的批间精密度变异系数为11.78%。我们确信所建立的方法具有高度可重复性，适用于高通量样本检测以确定不同肿瘤实体中的ARID1A浓度。将该ELISA技术应用于各类肿瘤及对照组织，将有助于探究肿瘤样本中ARID1A的定量检测是否具有预测价值。
肿瘤(癌症)患者之家