用于非小细胞肺癌耐药相关microRNAs联合检测的开关型电化学生物传感新方法的研究

Lung cancer is a malignant tumor of the highest morbidity and mortality which is a serious threat to human health. Chemotherapy is the main treatment of patients with lung cancer, however, patients will eventually develop resistance to the chemotherapeutics. For most, the prognosis of lung cancer is poor leading to high mortality. MiRNAs play a significant role in the regulation of drug metabolism. Furthermore, miRNAs can indicate the drug resistance of tumor cells to chemotherapy drugs. By detecting the associated miRNAs of drug resistance for lung cancer, we can foresee the sensitivity and possible resistance on chemotherapeutics which is beneficial to screen for the individualized treatment for patients in order to improve therapeutic efficiency. The uniform and ordered nanoporous gold modified Au multichannel electrode is prepared by square wave voltammetry in two steps from low to high frequency on Au electrode.This project proposes a multichannel array detector basing on switch mode technology with high sensitivity, high specificity and strong flux for the determination of miRNAs(miRNA-21、-155、-126 and let-7a-2) associated with drug resistance in non-small cell lung cancer. Combining with the gene hybridization and enzyme immunoassay, novel grapheme-Au nanocomposites are applied to load more enzyme with the help of duplex-specific nuclease enzyme, thus hybridization can be done with a loop progress. This method will fulfil the simultaneous detection of multiple miRNAs in plasma, warning drug resistance of lung cancer and it is of great significance for overcoming the patients’drug resistance and the choice of clinical individualized treatments for lung cancer to improve the survival.

肺癌发病率和死亡率位列恶性肿瘤首位，化疗是肺癌患者主要治疗方式，但肺癌细胞对化疗药物极易产生耐药，使肺癌死亡率高居不下。miRNAs可调控药物在体内的代谢，指示肿瘤细胞对抗癌药物耐药性。通过检测肺癌相关miRNAs，可预测患者对化疗药物的敏感性和耐受性，有利于筛选适合患者的个体化治疗方案以提高疗效。本项目在前期构建多通道电化学传感器检测RNA的基础上，以非小细胞肺癌耐药相关miRNAs (mi-21、-155、-126和let-7a-2)为检测对象，制备界面均一的纳米多孔金多通道阵列电极，结合杂交技术和酶联信号技术，采用石墨烯-金纳米复合材料连接酶催化放大信号和DSN酶对目标miRNA循环杂交，构筑敏感度高、特异性强、高通量开关型纳米多孔金多通道阵列传感器，实现血浆中多miRNAs同时检测，预警肺癌耐药，对肺癌患者临床个体化治疗方案的选择、避免和克服获得性耐药、延长患者生存期具有重大意义。

肺癌发病率和死亡率位列恶性肿瘤首位，化疗是肺癌患者主要治疗方式，但肺癌细胞对化疗药物极易产生耐药，使肺癌死亡率高居不下。miRNAs可调控药物在体内的代谢，指示肿瘤细胞对抗癌药物耐药性。通过检测肺癌相关miRNAs，可预测患者对化疗药物的敏感性和耐受性，有利于筛选适合患者的个体化治疗方案以提高疗效。本项目在前期构建多通道电化学传感器检测RNA的基础上，以非小细胞肺癌耐药相关miRNAs (mi-21、-155、-126和let-7a)为检测对象，制备界面均一的纳米铂颗粒/纳米多孔金多通道阵列电极，结合催化发卡组装技术，实现对目标miRNA循环杂交，构筑出敏感度高、特异性强、高通量开关型纳米多孔金多通道阵列传感器，同时实现血浆总RNA提取物中多miRNAs同时检测，并与临床检验金标准RT-PCR方法相比，测定结果基本相符，验证建立方法的准确性。该方法可实现肺癌耐药预警，对肺癌患者临床个体化治疗方案的选择、避免和克服获得性耐药、延长患者生存期具有重大意义。