构建仿生肾癌微流控芯片模型研究miR-21-PDCD4-AP-1反馈环路参与肾癌侵袭、转移的作用机制

The therapy of metastasis renal carcinoma is difficulty for tumor treatment of urinary system. Metastasis is closely related to the change of microenviroment. The tumor microenviroment ,which supports the merisis of tumor, has a complex relation with the tumor invasion and metastasis. At the same time, tumor cell remoulds tumor microenviroment constantly. It is proved by prior research group that PDCD4 plays an important role in renal tumor invasion and metastasis. But there are still so many doubts about how the PDCD4 taking effect by a downstream molecule passageway. According to the interreaction character of microenviroment where cell living ,the author designs and builds a bionic renal cell carcinoma metastasis model using microfluidic chips tissue engineering to real simulating renal carcinoma cell and observing the whole placation of renal cell carcinoma invasion and metastasis. Using this microfluidic chips, we study the molecular mechanism of miR21-PDCD4-AP-1 feedback loop in renal carcinoma cell metastasis. The purpose of this study was to explore a new method of reaching renal carcinoma cell metastasis and discussion molecular mechanism of miR21-PDCD4-AP-1 feedback loop in renal cell carcinoma metastasis and providing a larvaceous target spot that can be used in treating renal cell carcinoma metastasis. This study has vital scientific significance and favourable innovation.

转移性肾癌的治疗是泌尿系肿瘤治疗的难点，肾癌细胞发生转移的微环境变化与转移的发生密切相关，肿瘤的微环境支持着肿瘤的生长并且与肿瘤的侵袭、转移有着极其复杂的关系，同时肿瘤细胞对微环境也在进行不断的改造，前期课题组证实miR-21靶向PDCD4的在肾癌的侵袭、转移过程中起着重要的作用，但是PDCD4如何通过下游分子通路起作用的确切机理尚不清楚，本研究拟利用微流控芯片技术，依据体内细胞生存的微环境相互作用的特性，设计和构建一个仿生肾癌芯片转移模型，模拟体内肾癌细胞的真实状况，动态实时观察肾癌细胞侵袭和转移的全过程，并依托此平台研究miR-21-PDCD4-AP-1反馈环路在肾细胞癌转移中的分子机制。旨在建立肾癌细胞侵袭、转移的研究新方法，探讨miR-21-PDCD4-AP-1反馈环路在肾癌侵袭、转移中的分子机制，并为治疗肾癌转移提供新的潜在的靶点。有着重要的科学意义和良好的创新性。

肾癌是第三常见的泌尿系肿瘤。大约20 - 30%的肾癌患者在诊断时就已经发生转移，从诊断远处转移开始，患者的平均存活时间约为24个月，少于20%的患者存活5年以上。 在正常细胞系（HK-2）肾癌细胞系（786-O、Caki-1、A498）以及原代细胞中调节miR-21，通过qPCR验证转染效果(p<0.05)，通过Western blot显示PDCD4的表达随着miR-21的下调而增高(p<0.05)，同时肾癌细胞的侵袭能力下降。本实验利用微流控芯片模型观察细胞侵袭过程，该装置提供了一种快速、廉价的研究RCC侵袭机制的新平台。 研究表明，miR-21促进细胞株和原发癌肾细胞癌细胞侵袭，提示miR-21可能在肾肿瘤治疗的新靶点。本研究选取两种肾癌细胞株（786-O、A498）研究miR-21-PDCD4-AP-1反馈环路在肾细胞癌侵袭、转移的分子机制。通过PDCD4 siRNA转染实验所用细胞株（786-O、A498），观察转染前后的侵袭及迁移情况，并且提取其RNA，蛋白，用qRT-PCR及western blot对其转染效率进行分析及对其相关指标的进行检测，并为治疗肾癌转移提供新的潜在的靶点。