蛋白组MRM-MS与基因组HTS-ctDNA联合应用对肺癌个体化治疗与愈合评估价值的探索

There might be hundreds of susceptibility mutations exist in a single cancer cell. The efficiency in synergistic inhibition of the activity among multiple mutation points is the key factor to guarantee the therapeutic effect of any inhibitor. Therefore, a personalized treatment is critical for curing lung cancer patients in clinic. Recently, high-throughput sequencing of circular tumor DNA (ctDNA) , which is convenient and efficient to test multi-genetic slices and multi-mutation points, have been used for molecular pathological diagnosis and individualized treatment. This technique has shown a great diagnosis potential for cancer with high sensitivity. Mass spectrometry (MALD-TOF- MS/MS) related proteomic study also have the ability in simultaneous testing of tens or even hundreds of cancer biomarker with high convenience and efficiency. This technique can be used not only for cancer early diagnosis, but also for evaluation of therapeutic effect and after the cure. In the study of Lewis lung cancer (LLC), our data suggested that TRPM8 induces UCP2 to trigger metabolic transformation, whereas TRPA1 induces autophagy during adverse conditions. The combination of both genes contributes directly to an invasive phenotype in lung carcinoma. Our other result also indicated the high relationship of accuracy, sensitivity and specificity of the support vector machine (SVM) diagnostic model, indicating the important reference value of the model for determining the degree of solitary pulmonary nodules (SPN) differentiation and is suitable for the auxiliary diagnosis of benign and malignant SPN. Under refined classification of the medical examination of SPNS and routine lab test, our project is designed to collect the samples of blood and pathological biopsy from lung cancer patients at different stages of tumor progress. Then the changes of biomarker level in serum will be traced and the value of clinical diagnostic potential for lung cancer development will be determined. Our research goal is to monitor the potential metastasis of lung cancer tendency and to establish the individualized treatment measures in order to provide practical monitoring system with advanced technology in clinic.

癌细胞存在数百个基因突变点，多个突变点之间活性的协同抑制是控制肿瘤的关键，所以基因水平上治疗个性化方案的调整对肺癌的疗效很重要。高通量测序肿瘤循环DNA（ctDNA）具有同时测定多片段、多位点、快捷高效的特点，对肿瘤检测的高度敏感性，可用于肿瘤的分子病理学诊断及个性化用药指导，已在最近的肝癌监测研究中显示出巨大的诊断潜力。质谱相关的蛋白组研究也具有同时测定百种肿瘤分子标志物的快捷高效的特点，不仅适合早期诊断，也适用于评估临床疗效及预后状况。最近建立的分离肺结节(SPN) 模型显示，支持向量机在确定SPN分化的程度及区分良性及恶性具有明显优势。本项目拟以肺癌临床血液与病理活检为样本，通过SPN及常规医疗分组疾病发展的不同阶段点，检测ctDNA水平，确定其对监测肺癌转移倾向及个性化治疗调节的价值；测定蛋白组标志物的质谱变化，确定肿瘤疗效及预后评估的高科技检测手段。

我们以河南大学第一附属医院肺癌及肾癌临床血液与病理活检为样本，通过SPN及常规医疗分组疾病发展的不同阶段点，检测LncRNA水平，并证明HMGB3, RAB25, SCAMP3三个因子对及对癌肺癌及乳头状肾细胞转移倾向监测、个性化治疗调节的价值。在此基础上，发现并建立了腺嘌呤诱导的大鼠肾衰模型，通过硫化氢进行肿瘤与肾衰在氧化还原过程中的关系探讨，并第一次证实了硫化氢通过ROS/MAPK及NF-κB双重通路阻止凋亡与炎性过程改善大鼠的肾衰状态，为肺及肾癌早期发育发展、发病过程的揭示以及治疗方法的实施策略提供了重要的启示性依据及动物学模型。..肺肿瘤表面的ICAM-1表达水平和肺癌的发生、转移关系密切，L-选择素促进可溶性L-选择素的形成。肺癌组病理类型腺癌患者的ICAM-1和L-选择素阳性率均显著高于鳞癌患者，TNM分期Ⅲ+Ⅳ期患者的ICAM-1和L-选择素阳性率均显著高于Ⅰ+Ⅱ期（P<0.05），从而确立ICAM-1和L-选择素水平与肺癌发展、预后密切相关。另外对HDAC9基因沉默介导Wnt信号通路对非小细胞肺癌细胞生物学特性的作用机制，对HDAC9、JNK、β-catenin、Wnt-5的mRNA表达水平及HDAC9、JNK的蛋白表达水平进行测试，检测各组细胞转染后增殖能力、迁移能力、侵袭能力与凋亡情况，证实了HDAC9沉默表达可抑制Wnt信号通路，进而抑制非小细胞肺癌细胞的增殖、迁移与侵袭并促进细胞凋亡。另通过探讨CXC趋化因子配体-5（CXCL5）对肺癌肿瘤免疫的影响，证实肺癌患者的血清CXCL5水平显著高于感染性肺部疾病患者和健康者，肿瘤组织中CXCL5与PD-1/PD-L1水平均显著高于癌旁正常组织，说明CXCL5可能通过激活PD-1/PD-L1信号通路抑制肿瘤免疫进而促进肺癌的恶化。..基于 “热量控制”促进肺癌患者的正常细胞对抗高剂量的化疗、增强化疗效果的理论，建立了14天完全断食状态的特殊方法（成功率提高到48%），比较了断食7天与14天在整体效果上的差别，发现断食7~9天出现了明显转折，各项指标向未断食的对照组回归，说明了延长断食会激发人体潜能，对代谢综合征及肺癌治病因子的清除巨大潜在作用。未来将伴随一系列用于代谢综合征、脂肪控制及抗癌手段的开发和研讨。