太白贝母提取物对人结肠癌SW480细胞增殖抑制作用及其机制的研究

Inhibitory Effects and Its Possible Mechanisms of Total Alkaloids Extracted from Fritillaria Taipaiensis on the Proliferation of Human Colon Adenocarcinoma Cell Line SW480 in vitro

目的研究太白贝母总生物碱提取物对人结肠癌SW480细胞的增殖抑制作用及机制。方法太白贝母总生物碱提取物作用于人结肠癌SW480细胞,分为空白细胞组、0.25%DMSO溶剂处理组、太白贝母总生物碱提取物处理组(含总生物碱浓度分别为36.9 mg·L^(-1)、49.2 mg·L^(-1)、61.5 mg·L^(-1))。采用MTT试验、流式细胞术检测细胞周期,倒置光镜及透射电镜等观察各组细胞的增殖抑制情况、细胞周期变化、细胞显微和亚显微结构变化。结果与0.25%DMSO溶剂处理组比较,61.5 mg·L^(-1)总生物碱提取物处理细胞24 h、48 h、72 h后,细胞生长抑制呈现较为明显的时间依赖性;含太白贝母总生物碱36.9 mg·L^(-1)、49.2 mg·L^(-1)、61.5 mg·L^(-1)的提取物分别处理细胞48 h,细胞生长抑制呈现显著的剂量依赖性;流式细胞术PI单染结果显示,36.9 mg·L^(-1)、49.2 mg·L^(-1)太白贝母总生物碱提取物可将细胞阻滞于G2和S期,61.5 mg·L^(-1)太白贝母总生物碱提取物可将细胞阻滞于G2和G1期;61.5 mg·L^(-1)太白贝母总生物碱提取物处理48 h,细胞显微和亚显微结构发生显著坏死样变化。结论碱化热回流法提取的太白贝母总生物碱提取物具有明显的抗结肠癌SW480细胞活性,阻滞细胞周期进程为其机制之一。

Objective To investigate the anti-tumor effects of total alkaloids extracted from Fritillaria taipaiensis P. Y. Li on human colon adenocarcinoma cell line SW480 in vitro. Methods We investigated the inhibition of cell proliferation, cell cycle, cell microstructure and ul- trastructure changes on hmnan colon cancer SW480 cells among blank cell group, 0.25% DMSO group, 36.9 mg.L -1 total alkaloids group, 49.2 mg.L-1 total alkaloids group, 61.5 ntg.L-1 total alkaloids group before and after treated with hot reflux extraction from Fritillaria taipaiensis P. Y. Li, by using MTT, flow cytometry, inverted optical microscope and transmission electron microscope observation. Results Compared with cells treated with 0.25% DMSO solvent, the extract treatment group containing total alkaloids 36.9 mg.L 1, 49.2 mg.L 1, 61.5 mg.L i front Fritil- laria taipaiensis P. Y. Li could inhibit the proliferation of SW480 cells in different degrees at 24 h, 48 h, 72 h. When treated by 61.5 mg.L i total alkaloids, the proliferation rate of SW480 cells showed an obvious time-dependent relationship. When treated for 48 h, the proliferation rate of SW480 cells showed a significant dose-dependent relationship. The extract treatment group containing 36.9 mg.L i or 49.2 mg.L i total alkaloids inhibited cell proliferation by G2 and S phase arrest. However, the extract treatment group containing 61.5 nlg.L1 total alkaloids blocked the cell cycle in G2 and G1 phase. Necrosis-like changes were found in microstructure and ultrastructure of SW480 cells after treated by 61.5 mg.L-1 total alkaloids at 48h. Conclusion The total alkaloids extracted from Fritillaria taipaiensis P. Y. Li by heat reflux method had obvious anti-tumor effects on colon cancer SW480 cells, and blocking the cell cycle progression may be one of the mechanisms of inhibiting proliferation.