从Cyr61调控上皮间质转化角度探讨清胰化积方抗胰腺癌肝转移作用的分子机制

Liver metastasis prognosis is poor and the main death cause of pancreatic cancer，lack of effective treatment now. Group clinical study found that pancreatic cancer with liver metastasis patients treated by Qingyihuaji formula are significantly better than the control group；experimental study also revealed that qingyihuaji formula inhibit the growth of liver metastasis of pancreatic cancer cells and down-regulate Cyr61,but its mechanism remains to be further explored. Therefore,we propose a hypothesis:"hot and humid poison accumulation " is the core of pancreatic cancer pathogenesis and the key of metastasis, liver metastasis of pancreatic cancer treated by Qingyihuaji formula maybe down-regulate Cyr61 and reverse EMT expression. To test this hypothesis, we intend to use high liver metastatic cell SW1990HM as well as primary cell and nude mouse liver metastasis model,using Real time PCR,Western blot,lentiviral vector and RNA interference methods,to observe Qingyihuaji formula affecting Cyr61 regulation of EMT expression on the molecular,cellular,organization and the overall animal level, clear the molecular mechanism of anti-tumor metastasis. From the new point of view of the Cyr61 regulation of EMT, this study lay the foundation for revealing the mechanism of liver metastasis of pancreatic cancer treated by Chinese herbal compound, and provide new idea and new method of anti-tumor metastasis of Chinese medicine.

肝转移是胰腺癌预后差和主要致死原因之一，目前也缺乏有效的治疗手段。课题组临床研究发现以清胰化积方为主治疗胰腺癌肝转移患者疗效显著优于对照组；实验研究也揭示该方能抑制胰腺癌细胞肝转移生长以及下调Cyr61，但其作用机制还有待进一步探讨。故此，我们提出假说:"湿热毒聚"是胰腺癌发病的核心病机和转移的关键, 针对该病机的清胰化积方抗胰腺癌肝转移作用可能通过下调Cyr61逆转EMT表达。为了验证该假说，我们拟通过人胰腺癌高肝转移细胞SW1990HM以及原代细胞和肝转移裸小鼠模型，采用Real time PCR、Western blot、慢病毒载体和RNA干扰等手段，在分子、细胞、组织和动物整体水平上观察清胰化积方作用Cyr61调控EMT表达，明确该方抗肝转移作用的分子机制。本研究将从Cyr61调控EMT这个新视点为揭示中药复方抗胰腺癌肝转移的作用机制奠定基础，为中药治疗肿瘤转移提供新思路和新方法。

背景：肝转移是胰腺癌预后差和主要致死原因之一，目前也缺乏有效的治疗手段。课题组临床研究发现以清胰化积方为主治疗胰腺癌肝转移患者疗效显著优于对照组；实验研究也揭示该方能抑制胰腺癌细胞肝转移生长以及下调Cyr61，但其作用机制还有待进一步探讨。故此，我们提出假说:“湿热毒聚”是胰腺癌发病的核心病机和转移的关键, 针对该病机的清胰化积方抗胰腺癌肝转移作用可能通过下调Cyr61逆转EMT表达。.主要研究内容：Cyr61水平在胰腺癌患者中显著高于同一病人的癌旁组织；过表达 Cyr61可增强胰腺癌细胞增殖，然而抑制Cyr61下降胰腺癌细胞增殖；进一步分析表明，CYR61似乎激活磷脂酰肌醇3-激酶（PI3K）而不是外相关激酶/丝裂原活化蛋白激酶（ERK）胰腺癌细胞信号通路，从而诱导一个主要的细胞周期抑制剂P27核排斥增加细胞增殖。通过胰腺导管注射腺病毒载体携带的短发夹RNA（shRNA）Cyr61可以明显减少胰腺肿瘤切除小鼠的远处转移。而且，Cyr61耗尽的胰腺癌细胞明显抑制体外肿瘤球的形成，显著降低肿瘤皮下移植瘤的生长，显著降低过继移植到NOD/SCID小鼠成瘤的发生率。.重要结果：Cyr61促进胰腺癌细胞生长是通过激活PI3K/Akt信号通路增强P27核排斥，Cyr61阳性肿瘤干细胞促进胰腺癌远处转移。.结论：揭示胰腺癌Cyr61调节胰腺癌细胞增殖的分子基础，抑制胰腺癌肿瘤干细胞Cyr61表达可能会抑制原发肿瘤切除后的肿瘤细胞转移，突出Cyr61为胰腺癌治疗的新靶点。.