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目的:利用细胞瞬时转染技术,探讨过表达PDK1和Akt后化瘀解毒方抑制人胃癌SGC-7901细胞侵袭转移的机制。方法:先后将SGC-7901细胞瞬时转染PDK1质粒和Akt质粒,采用matrigel基质胶观察细胞黏附作用;利用Transwell细胞培养系统检测细胞迁移、侵袭作用;利用免疫共沉淀观察PDK1和Akt的结合力。结果:过表达Akt可以阻止化瘀解毒方提取物诱导的SGC-7901细胞黏附能力、迁移、侵袭能力下降。免疫共沉淀显示化瘀解毒方提取物通过抑制PDK1和Akt的结合进而抑制Akt磷酸化。结论:化瘀解毒方抑制胃癌细胞黏附、迁移、侵袭,其作用机制与抑制PI3K/Akt通路有关。
Objective: To investigate the mechanism of Huayu Jiedu Decoction inhibiting the invasion and metastasis of human gastric cancer cell line SGC-7901 by transient transfection of cells. Methods: SGC-7901 cells were transiently transfected with PDK1 plasmid and Akt plasmid, and matrigel matrices were used to observe cell adhesion. Transwell cell culture system was used to detect cell migration and invasion. The co-expression of PDK1 and Akt was observed by immunoprecipitation. Results: Overexpression of Akt could prevent the cell adhesion of SGC-7901 cells induced by Huayu Jiedu extract and decreased the ability of migration and invasion. Co-immunoprecipitation showed that Huayu Jiedu Fang extract can inhibit Akt phosphorylation by inhibiting the binding of PDK1 and Akt. Conclusion: Huayu Jiedu Fang inhibits the adhesion, migration and invasion of gastric cancer cells, and its mechanism is related to the inhibition of PI3K / Akt pathway.