基于NFAT5/S100A4/GDF15信号轴探讨益气健脾法对胃癌细胞干样特性的影响及其表观遗传学调控机制

Gastric cancer is the most common malignant tumor of digestive system. The recurrent and metastasis of gastric cancer are important factors affecting long-term survival in patients. Gastric cancer stem cells are closely related to the recurrence and metastasis of gastric cancer. In recent years, Traditional Chinese Medicine (TCM) received more and more attention in prevention and cure of malignant tumor. The strengthening spleen qi is the most common one of the TCM treatment methods for gastric cancer. Our previous study confirmed that NFAT5/S100A4/GDF15 signaling axis can influence the stem-like properties of gastric cancer cells MGC803. Strengthening spleen qi basic formula-sijunzi decoction can inhibit expression of that signaling axis molecules and the stem-like properties related genes in gastric cancer cells. It prompts that strengthening spleen qi can inhibit stem-like properties of cell to resist gastric cancer. In our study, we will treat human gastric cancer cells with serum containing sijunzi decoction, and then test the expression of stem-like properties related genes, spheroid, soft-agar colony forming abilities, the frequency of potential molecular markers of gastric cancer stem cells and xenotransplantation. We will study the epigenetics mechanism of sijunzi decoction regulating NFAT5/S100A4/GDF15 signaling axis from the DNA, RNA and protein level, respectively. And we will observe sijunzi decoction on human gastric cancer orthotopic tumor model in nude mice. Expect to provide the basis for strengthening spleen qi in the treatment of gastric cancer and related application.

胃癌是消化系统最常见的恶性肿瘤，其复发转移是影响患者长期生存的重要因素，而胃癌干细胞与胃癌的复发转移密切相关。近年来中医药在肿瘤防治方面越来越受到关注，中医治疗胃癌的常用治法是益气健脾法。课题组前期研究证实，NFAT5/S100A4/GDF15信号轴能够影响胃癌细胞干样特性；益气健脾基础方—四君子汤可抑制胃癌细胞中上述信号轴分子以及干性相关基因的表达，提示益气健脾法能够从抑制细胞干样特性角度发挥抗胃癌作用。本课题拟采用四君子汤含药血清处理胃癌细胞，从干性基因表达情况、细胞成球率、软琼脂集落形成能力、胃癌干细胞潜在分子标志物阳性表达细胞比例以及裸鼠成瘤等方面研究四君子汤对胃癌细胞干样特性的影响；从DNA、RNA和蛋白层面分别阐明四君子汤调控NFAT5/S100A4/GDF15信号轴的表观遗传学机制；并观察四君子汤对人胃癌裸鼠原位移植瘤的影响，以期为益气健脾法在胃癌的治疗及相关应用提供依据。

胃癌是消化系统最常见的恶性肿瘤，其复发转移是影响患者长期生存的重要因素，而胃癌干细胞与胃癌的复发转移密切相关。近年来中医药在肿瘤防治方面越来越受到关注，中医治疗胃癌的常用治法是益气健脾法。课题组前期研究证实，NFAT5/S100A4/GDF15信号轴能够影响胃癌细胞干样特性；益气健脾基础方—四君子汤可抑制胃癌细胞中上述信号轴分子以及干性相关基因的表达，提示益气健脾法能够从抑制细胞干样特性角度发挥抗胃癌作用。本课题研究结果显示，采用四君子汤含药血清处理胃癌细胞后，细胞成球率、软琼脂集落形成能力、CD44+细胞亚群比例以及干性相关基因OCT4、SOX2、CD44 的表达均受到明显抑制；细胞的凋亡率增加而增殖力、上皮间质转化能力均显著降低。此外，四君子汤灌胃对裸鼠成瘤具有明显的抑制作用，且能够有效改变瘤组织中miRNA和mRNA表达谱的改变。提示，四君子汤可能通过影响多种基因表达进而参与调控胃癌细胞多种生物学功能进而影响胃癌的复发转移。这一结果能够为益气健脾法在胃癌的治疗及相关应用提供理论和实验依据。