摘要
目的 研究槐耳多糖通过调节索尼克刺猬-Gli家族锌指蛋白1(Shh-Gli1)信号通路对肺癌细胞A549增殖、凋亡和血管生成的影响.方法 将肺癌细胞A549分为对照组(正常培养)及低、中、高剂量实验组(2、5、10 μg·mL-1槐耳多糖).给药24 h后,以噻唑蓝法检测细胞增殖水平,以流式细胞术检测细胞凋亡水平,以比色法检测细胞中胱天蛋白酶3(caspase-3)酶活性,以蛋白质印迹实验检测细胞中血管内皮细胞生长因子(VEGF)、血管内皮细胞生长因子受体2(VEGFR2)、Shh及Gli1蛋白的表达水平.结果 对照组和低、中、高剂量实验组的细胞存活率分别为 100.00％、(73.13±3.86)％、(47.97±6.12)％和(30.73±6.67)％,细胞凋亡率分别为(0.72±0.04)％、(19.21±1.27)％、(29.48±0.92)％和(46.29±2.81)％,细胞中 caspase-3 酶活性分别为 1.00、1.33±0.08、1.71±0.09和2.28±0.14,VEGF蛋白相对表达水平分别为1.17±0.04、0.95±0.08、0.39±0.04 和 0.11±0.03,VEGFR2 蛋白相对表达水平分别为0.99±0.04、0.80±0.03、0.29±0.03 和 0.24±0.03,Gli1 蛋白相对表达水平分别为 1.22±0.04、1.07±0.04、0.60±0.04 和 0.41±0.05.对照组的上述指标与低、中、高剂量实验组相比,在统计学上差异均有统计学意义(均P＜0.05).结论 槐耳多糖可通过调节Shh-Gli1信号通路抑制肺癌细胞的增殖及血管生成,并促进其凋亡.
Abstract
Objective To investigate the effects of Huaier polysaccharides on proliferation,apoptosis,and angiogenesis in A549 cells by regulating the Sonic hedgehog-Gli family zinc finger 1(Shh-Gli1)signaling pathway.Methods A549 cells were divided into control group(normal culture)and low,medium,high dose experimental groups(2,5,and 10 µg·mL-1 Huaier polysaccharides).After 24 h of treatment,cell proliferation level was detected by methyl thiazolyl tetrazolium assay,cell apoptosis level was detected by flow cytometry,cysteine aspartic acid specific protease-3(caspase-3)enzyme activity in cells was detected by colorimetric assay,and the expression levels of vascular endothelial growth factor(VEGF),vascular endothelial growth factor receptor 2(VEGFR2),Shh and Gli1 proteins in cells were detected by Western blot.Results The cell survival rates of control group and low,medium,high dose experimental groups were 100.00％,(73.13±3.86)％,(47.97±6.12)％and(30.73±6.67)％,respectively;cell apoptosis rates were(0.72±0.04)％,(19.21±1.27)％,(29.48±0.92)％and(46.29±2.81)％,respectively;caspase-3 enzyme activity in cells were 1.00,1.33±0.08,1.71±0.09 and 2.28±0.14,respectively;the relative expression levels of VEGF protein in cells were 1.17±0.04,0.95±0.08,0.39±0.04 and 0.11±0.03,respectively;the relative expression levels of VEGFR2 protein in cells were 0.99±0.04,0.80±0.03,0.29±0.03 and 0.24±0.03,respectively;the relative expression levels of Gli1 protein in cells were 1.22±0.04,1.07±0.04,0.60±0.04 and 0.41±0.05,respectively.There were statistical significance of above indicators between control group and low,medium,high dose experimental groups(all P＜0.05).Conclusion Huaier polysaccharides can inhibit proliferation and angiogenesis of A549 cells and promote apoptosis by regulating the Shh-Gli1 signaling pathway.
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