乳香挥发油抑制人肝癌SMMC-7721细胞株增殖及诱导凋亡的作用

Effects of Boswellia carterii Volatile Oils on Proliferation and Apoptosis of Liver Cancer Cell SMMC-7721

目的探讨乳香挥发油体外抑制人肝癌SMMC-7721细胞株增殖及诱导凋亡的作用。方法使用四甲基偶氮唑蓝(MTT)法观察乳香挥发油对SMMC-7721的增殖抑制作用;通过吖啶橙染色,观察SMMC-7721的形态学变化;使用琼脂糖凝胶电泳法检测乳香挥发油对细胞DNA降解的影响;利用流式细胞术分析乳香挥发油诱导SMMC-7721凋亡的凋亡率及对细胞周期的影响;应用间接免疫荧光法观察凋亡调控基因bax和bcl-2蛋白的表达变化。结果乳香挥发油浓度为0.7、0.07、0.007、0.0007、0.00007mg.mL-1作用24h时,能抑制SMMC-7721的生长,并呈浓度依赖性。在浓度为0.07mg.mL-1时,作用48h或72h,电泳结果显示了DNA梯状条带,流式细胞仪检测到凋亡峰,且细胞被阻滞于G1期。间接免疫荧光法的结果表明,促凋亡蛋白bax的表达增强,抗凋亡蛋白bcl-2的表达无明显变化。结论乳香挥发油能抑制肝癌细胞株SMMC-7721的增殖,并可能通过上调线粒体内bax/bcl-2的表达比例诱导SMMC-7721细胞的凋亡,而且其诱导的凋亡具有细胞周期依赖性。

AIM：To study the effect of volatile oils from Boswellia carterii Birdwood on the proliferation and apoptosis in human liver cancer SMMC-7721 cells; METHODS： Effects of proliferation were detected on SMMC-7721 by volatile oils from B. carterii with MTT assay;and the morphologic changes of cell were observed using acirdine staining; DNA fragmentation was detected by agarose gel electrophoresis; inhibitory rate and the change of cell-cycle were assessed by Flow Cytometry（FCM） ; the change of expression of protein bax and bcl-2 were observed by volatile oils from B. carterii with indirect immunefluorescence microscope. RESULTS： Volatile oils from B. carterii could inhibit the growth of the concentration of 0.7, 0.07, 0.007, 0.0007, 0.00007 mg·mL^-1 for 24 h, which depended on the concentration. At the concentration of 0.07 mg·mL^-1 for 48 h or 72h, the ＂DNA ladder＂ was clearly observed by agarose gel electrophoresis, apoptosis peak was detected by FCM, and cause the GI phase block of SMMC-7721 cells. Importantly, up-regulating of apopotosis gene expression of bax was detected, while the protein level of bcl-2 have no obvious change. CONCLUSION： Volatile oils from B. carterii could inhibit the reproduction of liver cancer cells SMMC-7721, and may induce apoptosis of SMMC-7721 cells through up-regulating the expression of bax/bcl-2, while its induced apoptosis cells is cell-cycle dependent.