斑蝥酸钠对人肺癌LAC细胞周期的影响及分子机制的研究

Effect of Sodium Cantharidinate on Arresting Cell Cycle of Human Pulmonary Carcinoma Cell LAC and Its Mechanism

【目的】探讨斑蝥酸钠对人肺癌LAC细胞周期的影响及其分子机制。【方法】取指数生长期的LAC细胞,分别加入2.5、12.5、25μg/mL的斑蝥酸钠和20μg/mL羟基喜树碱(HCPT),并以生理盐水作阴性对照,每组均设3个复孔;采用四甲基偶氮唑盐(MTT)法检测斑蝥酸钠对人肺癌LAC细胞增殖的抑制作用;采用流式细胞仪定量检测药物作用后肿瘤细胞凋亡及p53、bcl-2基因的蛋白表达。【结果】斑蝥酸钠具有显著性抑制人肺癌LAC细胞增殖的作用,其抑瘤效应优于HCPT。斑蝥酸钠作用于人肺癌LAC细胞后引起S期细胞明显减少,细胞被阻止于G0/G1期,部分细胞发生凋亡,效应优于HCPT(P<0.01)。经斑蝥酸钠作用后,LAC细胞的p53基因表达上调,而bcl-2基因表达下调。【结论】斑蝥酸钠抑制人肺癌LAC细胞增殖的作用可能与其能上调p53基因、下调blc-2基因的表达和诱导LAC细胞凋亡有关。

[ Objective ] To observe the effect of sodium cantharidinate （ SC ） on cell cycle of human pulmonary carcinoma cell LAC and to explore its molecular mechanism. [ Methods] LAC at logarithmic phase was cultured with 2.5, 12.5 and 25 μg/mL of SA and 20μg/mL of hydroxyl camptothecin （HCPT） respectively. Meanwhile, LAC cultured with saline served as the negative control. There were 3 double holes for each group. Methyl thiazolyl tetrazolium （MTT） assay was used for the detection of LAC proliferation, and flow cytometer was used for the detection of LAC apoptosis as well as p53 and bcl-2 gene expression. [ Results ] The inhibitory effect of SC on LAC proliferation was obvious and superior to HCPT. After culturing with SC, the cell count of LAC at S phase was reduced, indicating that LAC division was hold at G0/G1 phase and there existed apoptosis in some LAC （P 〈 0. 01 compared with HCPT） ; the p53 gene expression was increased and bcl-2 gene expression decreased in SC group. [ Conclusion] The mechanism of SC inhibiting LAC proliferation may be related with the up-regulation of p53 gene expression and down-regulation of bcl-2 gene expression, which contributes to LAC apoptosis.