青蒿琥酯对肝癌细胞生长的抑制作用及其机制

Inhibitory effect and its mechanism of artesunate on the growth of hepatoma cells

目的探讨青蒿琥酯(A RT)对肝癌细胞生长的抑制作用及其机制。方法以不同浓度青蒿琥酯作用肝癌SMMC-7721细胞24h,采用MTT法检测细胞生长抑制率(IC5 0)值。根据I C5 0值选取3个青蒿琥酯浓度(30、60、120μg/ml)进行后续实验。以不同浓度青蒿琥酯(0、30、60、120μg/ml)作用SM MC-7721细胞24h,0μg/ml青蒿琥酯以生理盐水代替青蒿琥酯作为对照组,分别命名为对照组、30μg/ml ART组、60μg/ml ART组、120μg/ml A RT组。倒置显微镜观察细胞形态;流式细胞术检测各组细胞周期、细胞凋亡、细胞线粒体膜电位及B c l-2和B a x蛋白表达水平。结果 MTT结果显示,青蒿琥酯对肝癌SMMC-7721细胞有生长抑制作用,且呈剂量依赖性,IC_(50)=60.27±1.51μg/ml。倒置显微镜下观察,青蒿琥酯作用24h后,SMMC-7721细胞中呈现不同程度的死亡细胞,且随着青蒿琥酯浓度增加,死亡破碎细胞也增加。30、60、120μg/ml ART组细胞凋亡率(分别为4.44%±0.32%、6.54%±0.84%、12.92%±1.22%)与对照组(2.15%±0.10%)相比明显增高(P<0.01)。30、60、120μg/ml ART组细胞周期G0/1期细胞数(分别为74.91%±0.69%、77.14%±1.39%、78.62%±0.55%)与对照组(70.60%±0.60%)相比明显增高(P<0.01),而30、60、120μg/ml ART组细胞增殖指数(PI)(分别为25.09%±0.69%、22.86%±1.39%、21.38%±0.55%)与对照组(29.40%±0.60%)相比明显降低(P<0.01)。30、60、120μg/ml ART组细胞线粒体膜电位(分别为244.36±3.27、229.51±4.74、173.17±6.93)与对照组(277.82±5.68)相比明显降低(P<0.01)。30、60、120μg/ml ART组细胞中Bcl-2蛋白表达(分别为220.86±7.76、195.64±9.23、148.32±8.19)与对照组(256.86±7.78)相比明显降低(P<0.01),而Bax蛋白表达(分别为214.22±2.01、231.54±6.16、260.96±11.38)与对照组(186.21±8.82)相比明显增高(P<0.01)。结论青蒿琥酯可抑制肝癌SMMC-7721细胞生长,其作用机制与诱导细胞凋亡及细胞周期阻滞有关。

Objective To investigate the inhibitor y effect of artesunate（ART） on hepatoma cells and its mechanism. Methods The growth inhibitory rate of SMMC-7721 cells was detected by MTT after the treatment of different concentrations of artesunate. IC50 value of artesunate on SMMC-7721 cell for 24 h was calculated. Three concentrations（30, 60, 120μg/ml） of artesunate were selected depending on the IC50 value. SMMC-7721 cells were divided into four groups： control group and 30, 60, 120μg/ml ART groups. Cell morphology was observed by inverted microscope and the cell cycle, cell apoptosis, mitochondrial membrane potential and the Bcl-2 and Bax protein expression levels in SMMC-7721 cells were determined by flow cytometr y after the treatment of different concentrations of ART（30, 60, 120μg/ml） for 24 h, while the same volume of saline was used in the control. Results The results of MTT showed that artesunate inhibited the growth of SMMC-7721 cells in a dose-dependent manner and IC50=（60.27±1.51）μg/ml. Different levels of cell death in SMMC-7721 cells were observed after the treatment of artesunate for 24 h, and with the artesunate concentration increasing, the death crushing cells also increased. The cell apoptosis rates of 30, 60, 120μg/ml ART groups（4.44%±0.32%, 6.54%±0.84%, 12.92%±1.22%） were significantly higher than that of control group（2.15%±0.10%, P〈0.01）. The G0/1 phase of 30, 60, 120μg/ml ART groups（74.91%±0.69%, 77.14%±1.39%, 78.62%±0.55%） were significantly higher than that of control group（70.60%±0.60%, P〈0.01）. The cell proliferation indices of 30, 60, 120μg/ml ART groups（25.09%±0.69%, 22.86%±1.39%, 21.38%±0.55%） were significantly lower than that of control group（29.40%±0.60%, P〈0.01）. The mitochondrial membrane potentials of 30, 60 and 120μg/ml ART groups（244.36±3.27, 229.51±4.74, 173.17±6.93） were significantly lower than that of control group（277.82±5.68, P〈0.01）. The Bcl-2 protein expression levels of 30, 60 and 120μg/ml ART groups（220.86±7.76, 195.64±9.23, 148.32±8.19） were significantly lower than that of control group（256.86±7.78, P〈0.01）. The Bax protein expression levels of 30, 60 and 120μg/ml ART groups（214.22±2.01, 231.54±6.16, 260.96±11.38） were significantly higher than that of control group（186.21±8.82, P〈0.01）. Conclusion Artesunate can inhibit the growth of hepatoma SMMC-7721 cells, and its mechanism is related to the induction of apoptosis and cell cycle arrest.