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目的筛选出槐耳浸膏与化疗药物的最佳组合并探讨槐耳浸膏和化疗药物5-氟尿嘧啶诱导人胃癌MGC803细胞凋亡的机制。方法以不同浓度槐耳浸膏和化疗药物(5-氟尿嘧啶,紫杉醇)作用于体外培养的人胃癌细胞MGC803,通过MTT法检测槐耳浸膏对MGC803细胞增殖的作用,倒置显微镜观察其形态的变化;将0.2 mg·m L-1的槐耳浸膏分别与5-FU(2.5μg·m L-1)或紫杉醇(0.5μg·m L-1)联合处理细胞,比较存活率。流式细胞术检测0.2 mg·m L-1槐耳浸膏联合5-FU(2.5μg·m L-1)干预MGC803细胞24、48 h后细胞的凋亡情况;Western Blot法检测0.2 mg·m L-1槐耳浸膏联合5-FU(2.5μg·m L-1)干预MGC803细胞24、48 h后细胞的P53、T-Akt和P-Akt蛋白的表达。结果槐耳浸膏及化疗药物(5-氟尿嘧啶,紫杉醇)均能降低MGC803细胞存活率,并呈时间和剂量依赖性;倒置显微镜下可见细胞胞体缩小、核碎裂,且呈浓度与时间依赖性(P<0.01)。相较于单独应用化疗药物,联合用药能够显著降低细胞的生存率。槐耳浸膏联合5-FU联合给药上调人胃癌细胞MGC803 T-Akt及P53蛋白表达量,同时下调P-Akt的表达量。结论槐耳浸膏联合5-FU在体外诱导胃癌细胞MGC803凋亡作用可能是通过抑制PI3K/Akt信号通路来实现的。联合使用槐耳浸膏和化疗药物可能会更有效发挥槐耳抗肿瘤增殖的作用。
Objective To screen out the best combination of the extract of Acacia confusa and chemotherapeutic drugs and to explore the mechanism of the apoptosis of human gastric cancer cell line MGC803 induced by 5-Fluorouracil. Methods Human gastric cancer cell line MGC803 was treated with different concentrations of Acacia confusa and chemotherapy drugs (5-fluorouracil and paclitaxel). The effect of Acacia confusa extract on the proliferation of MGC803 cells was detected by MTT assay. The morphological changes were observed by inverted microscope The cells were treated with 0.5 mg · mL-1 or 5-FU (0.5 μg · mL -1) or 0.5 mg · mL -1 paclitaxel (0.2 mg · mL -1) respectively, and the survival rates were compared. Flow cytometry was used to detect the apoptosis of MGC803 cells treated with 0.2 mg · mL -1 of Acacia confusa and 5-FU (2.5 μg · mL -1) for 24,48 h. Western Blot method was used to detect the cytotoxicity of 0.2 mg · mL -1 The expression of P53, T-Akt and P-Akt proteins in MGC803 cells treated with 5-FU (2.5μg · m L-1) m L-1 Acacia confusa and 24,48 h were investigated. Results Both the acacia extract and chemotherapeutic agents (5-fluorouracil and paclitaxel) could decrease the survival rate of MGC803 cells in a time and dose-dependent manner. The cell bodies were reduced and nuclear fragmentation was observed under inverted microscope, and the concentration and time-dependent (P <0.01). Compared with chemotherapy alone, combination therapy can significantly reduce the survival rate of cells. The combination of the Huai-er extract and 5-FU could up-regulate the expression of T-Akt and P53 and down-regulate the expression of P-Akt in human gastric cancer cell line MGC803. CONCLUSION: The effects of Acacia confusa extract combined with 5-FU on apoptosis of gastric cancer cell line MGC803 may be through inhibition of PI3K / Akt signaling pathway. Combined use of Huaer extract and chemotherapy drugs may be more effective in promoting the anti-tumor proliferation Huai Er role.