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目的考察毛蚶水提物体外抗肿瘤活性,研究其抗肿瘤作用机制。方法MTT法检测毛蚶水提物对9种人肿瘤细胞系生长的影响,并通过Giemsa染色光学显微镜下观察肿瘤细胞核的变化;流式细胞术及TUNEL染色检测毛蚶水提物对HT-29细胞周期的影响和凋亡情况;Western blotting检测凋亡相关蛋白Caspaes-3的表达情况。结果毛蚶水提物对人结直肠癌HT-29、HCT-116细胞,肝癌HepG-2、BEL-7402细胞及肺癌A549细胞具有较强的抑制作用,抑制作用呈作用时间和剂量依赖性。流式细胞术结合细胞Giemsa染色结果显示,毛蚶水提物能引起人结直肠癌HT-29细胞G2-M期阻滞,多核细胞增加。TUNEL染色显示毛蚶水提物有一定诱导细胞凋亡的作用。400μg.mL-1毛蚶水提物对HT-29细胞Caspase-3蛋白的表达具有下调作用。结论毛蚶水提物体外具有广谱抗癌活性;毛蚶水提物抑制人结直肠癌HT-29细胞生长的主要机制为诱导细胞凋亡及引起肿瘤细胞G2-M期阻滞,进而形成多核细胞,最终导致肿瘤细胞死亡。
Objective To investigate the antitumor activity of Mudan water extract in vitro and its anti-tumor mechanism. Methods MTT assay was used to detect the effects of aqueous extract from Mudanjian on the growth of nine human tumor cell lines. The changes of tumor nuclei were observed under light microscope with Giemsa staining. Flow cytometry and TUNEL staining were used to detect the cell cycle of HT-29 cells And the apoptosis of the cells. Western blotting was used to detect the expression of Caspaes-3. Results Muddy water extract on human colorectal cancer HT-29, HCT-116 cells, liver cancer HepG-2, BEL-7402 cells and lung cancer A549 cells have a strong inhibitory effect was time-and dose-dependent manner. Flow cytometry combined with Giemsa staining showed that Muddy water extract can cause G2-M phase arrest and multi-nuclear increase in human colorectal cancer HT-29 cells. TUNEL staining showed that Mudan water extract can induce cell apoptosis. 400μg.mL-1 Muddy water extract on the HT-29 cell Caspase-3 protein expression down-regulation. CONCLUSION: The water extract of Mochi has broad-spectrum anticancer activity in vitro. The main mechanism of aqueous extract of Mudanghutai inhibiting the growth of human colorectal cancer HT-29 cells is induction of apoptosis and G2-M arrest of tumor cells to form multi-nucleated cells Eventually leading to tumor cell death.