长链非编码RNAH19通过调控肿瘤细胞分泌IL-10和TGF-β参与肿瘤免疫逃逸的研究

Long non-coding RNA H19 increases in many tumor types. Secretion of suppressive cytokines IL-10 and TGF-β by tumor cells play important role in evading the immune attack. However, the relationship between these two events is never discovered. We found that H19 positively regulate the expression of TGF-β. Additionally, we have reported that H19 up-regulate the let 7 target genes by absorbing let 7 in cytoplasm, therefore, IL-10, the target of let 7 is also under the control of H19. So, our hypothesis is: H19 promote the expression of IL-10 and TGF-β via let 7 pathway therefore play a role in immune escape of tumor cells. To prove this hypothesis, siH19 will be used to silence the H19 in tumor cell line Hela and MBT. The H19 silenced mouse tumor cell line will be injected to immune compatible mice to form tumor. The effect of H19 on IL-10 and TGF-β will be evaluated in vitro and in vivo. The lymphocyte subpopulations, NK, Cytotoxic T cell(CTL), regulatory T cell (Treg), that infiltrate in tumors will be analyzed in amount and activity. The relationship between H19 and IL-10, TGF-β, let 7, Treg , CTL, NK will also be analyzed in clinic cervical cancer and bladder cancer samples. In this project, the mechanism of immune escape of tumor cells is studied from H19, a new angle of view. It will provide new knowledge on the biological meaning of H19 up-regulation in cancer and be benefit to breaking the immune tolerance of cancer cells.

长链非编码RNA H19在多种肿瘤组织表达升高。但其在肿瘤免疫逃逸中的作用从未被揭示过。我们的预实验表明H19正向调控TGF-β1的表达。此外，我们近期研究发现H19可通过吸收microRNA let 7，上调let 7的靶基因。因此，H19很可能也会上调IL-10（let 7的靶基因）的表达。由此，本项目的假设是：在肿瘤细胞中，升高的H19促进IL-10和TGF-β的产生，有利于肿瘤细胞免疫逃逸。本项目拟用siRNA技术使肿瘤细胞株的H19沉默，并用H19i沉默的细胞建立荷瘤小鼠模型，在体外和体内研究H19对肿瘤分泌IL-10和TGF-β的影响及机制，对调节性T细胞（Treg），杀伤性T细胞和NK细胞的趋化能力，数量和活性的影响。并在临床肿瘤样本中研究H19与上述因子及细胞的关系。本项目从H19这个新的视点研究肿瘤免疫逃逸的机制，深入了解H19在肿瘤的生物学意义，为破解肿瘤逃逸提供新思路