益气补肾方含药血清对人胃癌细胞SGC-7901增殖转移及CD44+EpCAM+干细胞比例的影响

Effects of Yiqi Bushen Formula-containing Serum on Proliferation and Metastasis of Human Gastric Cancer Cell SGC-7901 and CD44+EpCAM+ Double Positive Cancer Stem Cells Ratio

关键词:  益气补肾方  胃癌  增殖转移  干细胞

Key words:  Yiqi Bushen formula  gastric cancer  proliferation and metastasis  cancer stem cells

摘要:

目的 探讨益气补肾方含药血清对人胃癌细胞SGC-7901增殖转移能力及CD44⁺EpCAM⁺干细胞比例的影响。方法 分别以低、中、高不同浓度益气补肾方含药血清及空白血清干预人胃癌细胞SGC-7901。采用MTT法检测24，48，72 h细胞增殖抑制率，Transwell小室检测胃癌细胞迁移侵袭能力。采用5-氟尿嘧啶（5-Fu）共培养富集胃癌干细胞，流式细胞术检测药物干预后CD44⁺EpCAM⁺干细胞比例。结果 干预48，72 h后，益气补肾方组细胞增殖抑制率显著高于同时期阴性对照组及空白对照组，差异有统计学意义（P<0.05）。Transwell结果显示，与阴性对照组和空白对照组比较，益气补肾方各组透膜细胞数显著降低，迁移能力受明显抑制，差异有显著统计学意义（P<0.01）。流式细胞术结果显示，终浓度为20 μg·mL^-1的5-Fu共培养24 h后，胃癌干细胞比例显著提升，益气补肾方干预后各组细胞中CD44⁺EpCAM⁺干细胞比例均下调，中、高浓度组与模型对照组相比，差异有显著统计学意义（P<0.01）。结论 益气补肾方含药血清能显著抑制人胃癌细胞SGC-7901增殖，并呈时间、剂量相关性，其抗转移作用可能与下调胃癌干细胞表面标志物CD44、EpCAM表达，降低CD44⁺EpCAM⁺干细胞比例有关。

Abstract:

OBJECTIVE To investigate the effects of Yiqi Bushen Formula-containing serum on proliferation and metastasis of human gastric cancer cell SGC-7901 and CD44⁺EpCAM⁺ double positive cancer stem cells ratio. METHODS The human gastric cancer cell line SGC-7901 was intervened by low, medium and high concentrations of Yiqi Bushen Formula-containing serum and blank serum respectively. The proliferation inhibition rate of 24, 48, 72 h cells was detected by MTT assay. Transwell was used to detect the migration and invasion ability of gastric cancer cells. Gastric cancer stem cells were enriched by co-cultured with 5-fluorouracil (5-Fu). Flow cytometry was used to detect the CD44⁺EpCAM⁺ double positive cancer stem cells ratio in gastric cancer stem cells after drug intervention. RESULTS After 48, 72 h intervention, the cell proliferation inhibition rate of the Yiqi Bushen Formula group was significantly higher than that of the negative control group and the blank control group, with a significant difference (P<0.05). Transwell showed that compared with the negative control group and the blank control group, the number of permeable membrane cells in each group of Yiqi Bushen Formula group decreased significantly, and the migration ability was significantly inhibited, with a significant difference (P<0.01). Flow cytometry showed that after co-culture with 20 μg·mL^-1 5-Fu, the proportion of stem cells in gastric cancer increased significantly. The CD44⁺EpCAM⁺ double positive cancer stem cells ratio in each group was down. The middle and high concentration groups was higher than that of the model control group with a significant difference (P<0.01). CONCLUSION Yiqi Bushen Formula-containing serum can significantly inhibit the proliferation of human gastric cancer cell SGC-7901, and it is time and dose correlation. Its anti metastasis effect may be related to the down regulation of the expression of CD44 and EpCAM from the surface markers and reduction of the CD44⁺EpCAM⁺ double positive cancer stem cells ratio in gastric cancer cells.