紫草素对人胃癌细胞增殖和细胞周期的影响

Effect of shikonin on proliferation and cell cycle of human gastric cancer cells

目的探讨紫草素对人胃癌细胞增殖、细胞周期及第10号染色体缺失的磷酸酶和张力蛋白同源物基因(PTEN)、细胞周期蛋白D1(CyclinD1)表达的影响。方法体外培养人胃癌BGC-823细胞,低、中、高剂量实验组与对照组分别以4.0,8.0,16.0μg·mL^-1紫草素与等量生理盐水干预48 h。用细胞增殖/毒性检测(CCK-8)法测定人胃癌BGC-823细胞的增殖抑制率,用流式细胞术碘化丙啶(PI)染色法检测人胃癌BGC-823细胞周期,用蛋白免疫印迹法检测人胃癌BGC-823细胞PTEN、CyclinD1蛋白表达水平。结果低、中、高剂量实验组人胃癌BGC-823细胞的增殖抑制率分别为(34.51±2.97)%,(49.62±4.06)%和(63.84±4.73)%;对照组与低、中、高剂量实验组人胃癌BGC-823细胞G0/G1期的比例分别为(39.42±5.83)%,(50.65±5.72)%,(65.51±7.80)%,(76.11±11.02)%;S期比例分别为(40.46±7.12)%,(29.46±5.18)%,(14.57±3.96)%,(3.73±1.03)%;PTEN蛋白相对表达量分别为0.06±0.04,0.18±0.09,0.42±0.11,0.91±0.09;CyclinD1蛋白相对表达量分别为0.98±0.02,0.83±0.12,0.37±0.13,0.11±0.08。上述指标,低、中、高剂量实验组分别与对照组比较(增殖抑制率除外),差异均有统计学意义(均P<0.05)。结论紫草素可显著抑制人胃癌BGC-823细胞增殖,使细胞周期阻滞于G0/G1期,同时促进其凋亡,其机制可能与上调人胃癌BGC-823细胞PTEN蛋白表达和下调CyclinD1蛋白表达相关。

Objective To investigate the effect of shikonin on the proliferation,cell cycle and phosphatase and tensin homolog deleted on chromosome 10(PTEN)and CyclinD1 in human gastric cancer cells.Methods Human gastric cancer BGC-823 cells were cultured in vitro,the exp-L/-M/-H groups and the control group were intervened with 4.0,8.0,16.0μg·mL^-1 shikonin and the same amount of normal saline respectively,and intervened for 48 h.The proliferation inhibition rate of human gastric cancer BGC-823 cells was determined by cell counting kit-8(CCK-8);Flow cytometry propidium iodide(PI)staining method was used to detect the cell cycle of human gastric cancer BGC-823;Western blot method was used to detect the expression levels of PTEN and CyclinD1 protein in human gastric cancer BGC-823 cells.Results The proliferation inhibition rates of human gastric cancer BGC-823 cells in the exp-L/-M/-H groups were(34.51±2.97)%,(49.62±4.06)% and(63.84±4.73)%;The ratios of G0/G1 phase of human gastric cancer BGC-823 cells in the control group and the exp-L/-M/-H groups were(39.42±5.83)%,(50.65±5.72)%,(65.51±7.80)%,(76.11±11.02)%;The ratios of S phase were(40.46±7.12)%,(29.46±5.18)%,(14.57±3.96)%,(3.73±1.03)%;The relative expression levels of PTEN protein were 0.06±0.04,0.18±0.09,0.42±0.11,0.91±0.09;The relative expression levels of CyclinD1 protein i were 0.98±0.02,0.83±0.12,0.37±0.13,0.11±0.08.Comparing between exp-L/-M/-H groups and control group,the differences in the above factors(except proliferation inhibition rates)were significant(all P<0.05).Conclusion Shikonin can significantly inhibit the proliferation of human gastric cancer BGC-823 cells,block the cell cycle in G0/G1 phase,and promote its apoptosis,and the mechanism may be related to the up-regulation of PTEN protein expression and down-regulation of CyclinD1 protein expression in human gastric cancer BGC-823 cells.