半夏泻心汤对胃癌微环境中骨髓间充质干细胞恶性转化的干预作用及其配伍机制研究

Bone-marrow Mesenchymal Stem Cells (BMSCs)， being biological carrier for gastric cancer therapy, but in the microenvironment of gastric cancer, might differentiate disorderly, become malignant, lead to safy risk. Whether this can be intervened is not clear up to now.Clinical study shows banxiaxiexin decoction can get exact effect in the prevention and treatment of gastric cancer. Our previous research shows Pinelliae Decoction for Purging Stomach-fire can reverse the gastric precancer change, inhibit the proliferation of gastric cancer cells, and induce their apoptosis. We speculate that Pinelliae Decoction for Purging Stomach-fire can interact with the gastric cancerous microenvironment and intervene BMSCs' malignant transformation. To prove this hypothesis, we intend to co-culture BCG-823 cells and BMSCs to establish BMSCs' growth transformation model in the gastric cancerous microenvironment, observe its biological change and tumor formation in vivo, at the same time observe intervening role in BMSCs'malignant transformation and influence on the migration of the serum treated with Pinelliae Decoction for Purging Stomach-fire and its different combinations, and discuss its mechanism, focusing on its relationship with the combination. The subject will, from a new angle, explain the intervening role in the BMSCs' malignant transformation in the gastric cancerous microenvironment of banxiaxiexin decoction and its combination mechanism in order to provide new ideas and target site of drug for the secure application of BMSCs to the prevention and treatment of gastric cancer.

骨髓间充质干细胞(BMSCs)作为胃癌生物治疗的载体，在胃癌微环境中可分化失调，恶性转化，有安全风险，能否被干预，迄今并不清楚。临床研究表明半夏泻心汤防治胃癌，疗效确切。前期我们已证实，半夏泻心汤可逆转胃癌前病变，抑制胃癌细胞增殖，诱导胃癌细胞凋亡。我们推测半夏泻心汤能作用于胃癌微环境，有干预BMSCs恶性转化的作用。为证实该假说，我们拟将人胃癌BCG-823细胞与BMSCs共培养，建立胃癌微环境中BMSCs的生长转化模型，观察BMSCs的生物特性变化及体内成瘤情况，同时观察半夏泻心汤及其不同配伍药组含药血清对BMSCs恶性转化的干预作用及对迁移趋化特性的影响，并探讨其作用机制，重点关注与"配伍"的关系。本课题将从新的视觉阐明半夏泻心汤对胃癌微环境中BMSCs恶性转化的干预作用及其配伍机制，为BMSCs安全应用于防治胃癌提供新思路和药物作用的新靶点。

骨髓间充质干细胞(BMSC)作为胃癌生物治疗的载体，在胃癌微环境中可分化失调，恶性转化，有安全风险，能否被干预，迄今并不清楚。临床研究表明半夏泻心汤防治胃癌，疗效确切。前期我们已证实，半夏泻心汤可逆转胃癌前病变，抑制胃癌细胞增殖，诱导胃癌细胞凋亡。我们推测半夏泻心汤能作用于胃癌微环境，有干预BMSC恶性转化的作用。为证实该假说，我们将人胃癌BCG-823细胞与BMSC共培养，建立胃癌微环境中BMSC的生长转化模型，观察BMSC的生物特性变化及体内成瘤情况，同时观察半夏泻心汤及其不同配伍药组含药血清对BMSCs恶性转化的干预作用及对迁移趋化特性的影响，并探讨其作用机制，重点关注与“配伍”的关系。研究表明，骨髓间充质干细胞在胃癌微环境中细胞形态与胃癌有相似之处，与细胞增殖恶化调控相关的端粒酶(Telomerase) 活性基因c-myc 及端粒酶逆转录酶（Telomerase reverse transcriptase，TERT）表达水平明显升高，但干细胞标志物CD44仍呈阳性表达，有向胃癌干细胞恶性转化的趋势。转化后的BMSC G2/M期细胞比例显著增加，生长增殖率及侵袭迁移能力明显升高。裸鼠体内可形成肿瘤结节，电镜显示肿瘤结节具备了肿瘤组织的结构特征，且肿瘤结节组织中促癌基因潜伏膜蛋白LMP1呈高表达状态，抑癌基因TCF21呈低表达状态，具有体内成瘤性。半夏泻心汤及配伍药组可降低转化后胃癌相关干细胞的c-myc和TERT表达水平，抑制其生长增殖、侵袭迁移及体内成瘤，以半夏泻心汤全方配伍最为显著。本项目证实了BMSC在胃癌微环境中可向胃癌相关干细胞恶性转化，揭示了半夏泻心汤干预BMSC在胃癌微环境中向胃癌相关干细胞恶性转化的作用及机理。从新的视觉阐明半夏泻心汤对胃癌微环境中BMSCs恶性转化的干预作用及其配伍机制，为进一步研究胃癌干细胞及BMSCs安全应用于防治胃癌及提供新思路和药物作用的新靶点。