调控HO-1表达对肿瘤耐药的逆转作用及机理研究

HO-1在肿瘤组织中多呈高表达常伴随化疗耐药。本项目以多种肿瘤耐药细胞株及肿瘤干细胞和荷瘤小鼠为研究对象。采用慢病毒介导转基因手段和RNA沉默并结合诱导剂/抑制剂靶向调控HO-1表达。体内外实验初步结果显示，HO-1与肿瘤耐药呈高度相关，其机制涉及多条通路，下调HO-1可在体内外实验中增强肿瘤化疗敏感性。下一步拟将采用基因测序，蛋白芯片和real-time PCR高通量筛选通路等手段在体外探索HO-1与耐药的关系及其发生机制；动物实验中采用ELISA分析体内肿瘤微环境中炎性因子的变化，Hoechst示踪肿瘤化疗敏感性，免疫组化分析与HO-1耐药相关通路蛋白表达情况，试证实HO-1可作为肿瘤耐药相关的潜在靶点，下调HO-1可以逆转耐药，并阐明其耐药机制，为临床克服肿瘤耐药提供实验依据。

Resistance toward anti-cancer drugs in tumor is usually accompany with up-regulation of Heme Oxygenase-1 （HO-1）. In this study, various tumor cells lines resisted to chemotherapy, primary tumor stem cells and tumor-bearing mice were employed as research targets. lentivirus and the RNA interference technology plus inducer/inhibitor strategy combined with the gene transfer were used for the regulation of HO-1 expression. The primary results of our in-vitro study demonstrated that HO-1 was closely correlated with tumor drug-resistance in which multiple pathways were involved, and the down-regulation of HO-1 in ex-vivo experiment increased the sensitivity to chemotherapy. Then, gene-sequencing, protein-chip, real-time PCR high-throughput screening technology will be used in our following in-vitro study on the relationship between HO-1 and drug-resistance and also on the underlying mechanisms. In animal experiments, ELISA analysis will be taken for the detection of inflammatory molecule in tumor micro-environment, Hoechst tracing technology be used for chemotherapy sensitivity analysis, and immunohistochemistry be used for the expression analysis of HO-1 related pathways involved in drug-resistance, trying to elucidate the role of HO-1 as a potential therapeutic target for drug-resistance,down-regulating HO-1 might be able to reverse the drug-resistance,and to disclose the underlying mechanism. Accumulating experimental evidence for the study on drug-resistance in tumor therapy.