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目的:探讨氧化苦参碱(oxymatrine,OM)对人结肠癌LOVO细胞增殖和凋亡的影响及其可能的机制。方法:采用MTT法检测OM对LOVO细胞的增殖抑制率;流式细胞仪检测LOVO细胞凋亡率以及细胞周期分布;吉姆萨染色法观察用药前后细胞形态变化;荧光定量逆转录-聚合酶链式反应(qRT-PCR)法检测药物作用后p53 mRNA表达变化情况。结果:MTT法显示,OM在一定浓度范围(200μg/mL～5mg/mL)对人结肠癌LOVO细胞的生长有明显的抑制作用,对LPS诱导的LOVO细胞的过度增殖也具有明显的抑制作用。LOVO细胞经OM作用后光镜下可见细胞数量明显减少,细胞内空泡明显增多。流式检测表明OM可明显诱导LOVO细胞凋亡,使LOVO细胞周期停滞于S期。荧光定量PCR法显示,OM作用后LOVO细胞p53 mRNA表达明显下降。结论:OM能显著抑制人结肠癌LOVO细胞增殖,并诱导LOVO细胞凋亡,其机制可能与OM使LOVO细胞周期停滞于S期,下调LOVO细胞p53 mRNA表达有关。
Objective: To investigate the effects of oxymatrine (OM) on proliferation and apoptosis of human colon carcinoma LOVO cells and its possible mechanism. Methods: The inhibitory rate of OM on LOVO cells was detected by MTT assay. The apoptosis rate and cell cycle distribution of LOVO cells were detected by flow cytometry. The changes of cell morphology were observed by Giemsa staining. Fluorescence quantitative reverse transcriptase-polymerase chain reaction Response (qRT-PCR) method was used to detect the change of p53 mRNA expression. Results: MTT assay showed that OM could significantly inhibit the growth of human colon cancer LOVO cells in a certain concentration range (200μg / mL ~ 5mg / mL) and inhibit the proliferation of LOVO cells induced by LPS. After treated with OM, the number of cells in LOVO cells was significantly decreased and the number of vacuoles in cells was significantly increased. Flow cytometry showed that OM could obviously induce apoptosis of LOVO cells and arrest the cell cycle of LOVO in S phase. Fluorescent quantitative PCR showed that the expression of p53 mRNA in LOVO cells was significantly decreased after OM treatment. CONCLUSION: OM can significantly inhibit the proliferation of human colon cancer LOVO cells and induce the apoptosis of LOVO cells, which may be related to the fact that OM blocks the LOVO cell cycle in S phase and down-regulates p53 mRNA expression in LOVO cells.