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目的:探讨异常黑胆质成熟剂醇提物的抗癌作用机理。方法:采用四氮甲基唑蓝法(MTT)观察样品对人肝癌细胞株(HepG2)体外生长的抑制作用;采用琼脂糖凝胶电泳技术、流式细胞术等观察样品对HepG2细胞凋亡的影响;采用逆转录-多聚酶链式反应技术(RT-PCR)观察样品对HepG2细胞凋亡相关基因mRNA表达的影响;观察样品对HepG2细胞Caspase-3活性的影响。结果:异常黑胆质成熟剂醇提物可明显抑制癌细胞体外生长(P<0.05);明显阻滞癌细胞周期、诱导癌细胞凋亡(P<0.05);明显提高抑癌基因p53、p21基因mRNA的表达,抑制抗凋亡基因Bc l-2 mRNA的表达,对Bax基因mRNA表达不产生明显的影响;明显提高Caspase-3活性。结论:异常黑胆质成熟剂醇提物抗癌作用可能与其抑制癌细胞生长、阻滞癌细胞周期、诱导癌细胞凋亡、调节凋亡相关基因表达、激活Caspase-3活性等功能有密切联系。
Objective: To investigate the anticancer mechanism of alcohol extract of abnormal savda. Methods: MTT assay was used to observe the inhibitory effect of the sample on the growth of human hepatocellular carcinoma cell line HepG2. The apoptosis of HepG2 cells was observed by agarose gel electrophoresis and flow cytometry The influence of samples on the mRNA expression of apoptosis-related genes in HepG2 cells was observed by reverse transcription-polymerase chain reaction (RT-PCR). The effect of samples on the activity of Caspase-3 in HepG2 cells was observed. Results: Alcohol extract of abnormal savda could significantly inhibit the growth of cancer cells in vitro (P <0.05), significantly inhibit the cell cycle and induce the apoptosis of cancer cells (P <0.05), and significantly increase the expression of tumor suppressor genes p53, p21 The mRNA expression of Bcl-2 and anti-apoptotic gene Bcl-2 mRNA had no significant effect on the mRNA expression of Bax, but significantly increased the activity of Caspase-3. CONCLUSION: The anticancer effect of alcohol extract from abnormal savda may be closely related to its function of inhibiting the growth of cancer cells, arresting the cell cycle, inducing the apoptosis of cancer cells, regulating the expression of apoptosis related genes and activating the activity of Caspase-3 .