脾虚内环境OATP表达对小鼠肝癌生长及转移影响研究

The imbalance in the internal environment is closely related with the development of tumor. The barrier of transport and transformation of dampness which caused by spleen deficiency is an important reason for the internal environmental change. It is found that improving internal environment of spleen deficiency have been proven effective in prevention and treatment of tumors in clinic, but its biological basis is unclear. In a previous study, we found that the organic anion transporter polypeptide(OATP) expression in the environment of spleen deficiency was closely related to transport and transformation of dampness, and abnormal expression of OATP was observed in mice with hepatocellular carcinoma in the pre-experimental. We speculate that the abnormal expression of OATP in the environment of spleen deficiency might be related to the development of hepatocellular carcinoma. Proposed " the environment of - The barrier of transport and transformation of dampness - tumor related" hypothesis. To test this hypothesis, we will copy hepatocellular carcinoma model with spleen deficiency in mice, detect the expression of OATP subfamily OATP2A1, OATP4A1 OATP1B3 and OATP2B1 In the organs and tissues 0f mice using RT-PCR, Western-blot and immunohistochemistry and other techniques, observe the Relationship between OATP expression pattern and the growth and metastasis of hepatocellular carcinoma,explore the effect of he barrier of transport and transformation of dampness on the growth and metastasis of hepatocellular carcinoma, and to provide new therapeutic strategies in Treating hepatocellular carcinoma for Traditional Chinese Medicine, promote the improvement and development of traditional Chinese medicine in the theory of water metabolism.

内环境失衡与肿瘤的发生发展关系密切。脾虚湿浊转运障碍是机体内环境变化的重要原因。临床研究发现改善脾虚内环境对防治肿瘤有效，但其生物学基础尚不明确。在前期研究中，我们发现脾虚内环境有机阴离子转运肽(OATP)表达与湿浊转运密切相关，并在预实验中观察到脾虚小鼠肝癌组织OATP表达异常。我们推测脾虚内环境OATP异常表达可能与肝癌发生发展密切相关，提出"脾虚内环境- - 湿浊转运障碍-肿瘤相关"假说。为证实这一假说，我们将复制脾虚肝癌小鼠模型，采用RT-PCR、Western-blot和免疫组化等技术，检测OATP亚家族OATP2A1、OATP4A1、OATP1B3和OATP2B1在脾虚肝癌小鼠相关脏腑组织的基因表达、蛋白表达和蛋白分布，观察OATP表达规律与肝癌生长及转移的关系，探讨脾虚内环境湿浊转运障碍对肝癌生长及转移的影响，为中医药防治肝癌提供新的治疗策略，促进中医水液代谢理论的完善和发展。

脾虚内环境湿浊转运障碍是肝癌发生发展的重要原因,但其分子机理未明，在一定程度上阻遏了脾虚湿浊转运障碍相关性肝癌的防治。本项目通过造模脾虚肝癌小鼠模型，采用RT-PCR、Western-blot和免疫组化等技术，检测OATP亚家族OATP2A1、OATP4A1、OATP1B2和OATP2B1在脾虚肝癌小鼠相关脏腑组织的基因表达、蛋白表达和蛋白分布，发现：脾虚基础的原位移植肝癌小鼠模型和药物诱导肝癌模型的成瘤率、肿瘤体积显著高于对照组而生存时间显著降于对照组；脾虚上调了两组肝癌模型小鼠的肿瘤标记物如AFP、VEGF、PCNA和Ki-67的表达，提示脾虚在肝癌的发生发展中是一种不利因素，促进了肝癌进程。在脾虚状态存在的基础上实施原位肝癌移植模型和药物诱导肝癌模型后，Oatp亚家族在各组织的表达和分布较非脾虚肝癌组（原位移植组、药物诱导组）不同，具体表现为脾虚湿浊运化功能障碍，同时肝疏泄功能减弱和小肠分清泌浊功能障碍，进一步加剧了脾虚湿浊聚集于机体内。同时，结肠和肺则表现为脾虚肝癌状态下，湿浊和糟粕进一步聚集于局部的状态，为一种代偿性的湿浊转移的机体自我调节。脾虚状态下的肝癌小鼠与非脾虚肝癌小鼠的结局以及小鼠各组织Oatp亚家族的表达和分布各不相同，其效应并非单纯肝癌和单纯脾虚两种因素的简单相加，而是脾虚和肝癌两种因素共同对机体产生的协同效应。以上研究探讨了脾虚内环境湿浊转运障碍对肝癌生长及转移的影响，为中医药防治肝癌提供新的治疗策略，促进中医水液代谢理论的完善和发展。