You Zhimei, Chen Dilong, Wei Qiang, Zhao Liang, Xia Jing, Li Danyang, Li Jing
Laboratory of Stem Cell and Tissue Engineering, Department of Histology and Embryology, Chongqing Medical University, Chongqing 400016, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2014 Jun;30(6):565-8.
To investigate the effect of ginsenoside Rh2 on leukemia KG1-α cells.
KG1-α cells were cultured in 96-well plate and harvested in the exponential phase. The cells were induced in the presence of different concentrations of Rg1, Rb1, Rh2 respectively for 24, 48, 72 hours. The conventional culture was performed in blank control group, and cytarabine hydrochloride as the positive control. The inhibitory effects of ginsenoside Rb1, Rg1, Rh2 on the growth of KG1-α cells were tested by CCK-8 assay. Then the most efficient ginsenoside was chosen for the further study by IC50. The effects of the ginsenoside Rh2 on cell cycle and apoptosis were detected by flow cytometry (FCM) combined with PI staining and annexin V-FITC/PI, respectively. The expressions of P53, P21, cyclin D1 and cleaved-caspase-3 were examined by Western blotting.
CCK-8 assay results showed that IC50 of ginsenoside Rh2, ginsenoside Rb1, ginsenoside Rg1 and cytarabine were 75, 207, 268, 1058 μmol/L, respectively. Compared with those in blank control group, the KG1-α cells after treated with 75 μmol/L ginsenoside Rh2 showed up-regulated apoptosis rates from (5.37±0.02)% to (8.37±0.015)% at 24 hours and to (33.22±1.67)% at 48 hours (P<0.05). In addition, the percentage of cells in G0/G1 phase increased significantly from (26.78±3.14)% to (29.26±2.31)% at 24 hours and to (44.77±2.26)% at 48 hours, and the percentage in S phase decreased significantly from (65.43±2.22)% to (51.46±0.57)% and (48.29±1.80)%, respectively. The expression levels of cleaved-caspase 3, P53 and P21 proteins went up significantly when the cells were treated with 75 μmol/L Rh2(P<0.05), meanwhile cyclin D1 protein dropped significantly (P<0.05).
Ginsenoside Rh2 could inhibit the proliferation of KG1-α cell and prompt its apoptosis.
研究人参皂苷Rh2对白血病KG1-α细胞的作用。
将KG1-α细胞接种于96孔板,在指数生长期收获细胞。分别用不同浓度的Rg1、Rb1、Rh2诱导细胞24、48、72小时。空白对照组进行常规培养,盐酸阿糖胞苷作为阳性对照。采用CCK-8法检测人参皂苷Rb1、Rg1、Rh2对KG1-α细胞生长的抑制作用。然后通过IC50选择最有效的人参皂苷进行进一步研究。分别采用流式细胞术(FCM)结合PI染色和膜联蛋白V-FITC/PI检测人参皂苷Rh2对细胞周期和凋亡的影响。通过蛋白质印迹法检测P53、P21、细胞周期蛋白D1和裂解的半胱天冬酶-3的表达。
CCK-8法检测结果显示,人参皂苷Rh2、人参皂苷Rb1、人参皂苷Rg1和阿糖胞苷的IC50分别为75、207、2六百六十八、1058μmol/L。与空白对照组相比,75μmol/L人参皂苷Rh2处理后的KG1-α细胞在24小时时凋亡率从(5.37±0.02)%上调至(8.37±0.015)%,48小时时上调至(33.22±1.67)%(P<0.05)。此外,G0/G1期细胞百分比在24小时时从(26.78±3.14)%显著增加至(29.26±2.31)%,48小时时增加至(44.77±2.26)%,S期细胞百分比分别从(65.43±2.22)%显著降低至(51.46±0.57)%和(48.29±1.80)%。当细胞用75μmol/L Rh2处理时,裂解的半胱天冬酶3、P53和P21蛋白的表达水平显著升高(P<0.05),同时细胞周期蛋白D1蛋白显著下降(P<0.05)。
人参皂苷Rh2可抑制KG1-α细胞增殖并促进其凋亡。