| 郭慧,刘鹏飞.人参皂苷 Rg5通过抑制蛋白激酶 B信号通路调控肝癌 HepG2细胞生物学行为[J].安徽医药,2021,25(6):1095-1099. |
| 人参皂苷 Rg5通过抑制蛋白激酶 B信号通路调控肝癌 HepG2细胞生物学行为 |
| Ginsenoside Rg5 regulates the biological behavior of hepatocellular carcinoma HepG2 cells by inhibiting AKT signaling pathway |
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| DOI：10.3969/j.issn.1009-6469.2021.06.008 |
| 中文关键词: 肝肿瘤 HepG2细胞 人参皂苷 Rg5 细胞增殖 侵袭 迁移 细胞周期 凋亡 AKT信号通路 |
| 英文关键词: Liver neoplasms HepG2 cells Ginsenoside Rg5 Cell proliferation Invasion Migration Cell cycle Apoptosis AKT signaling pathway |
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| 中文摘要: |
| 目的探讨人参皂苷 Rg5对肝癌 HepG2细胞生物学行为和蛋白激酶 B(AKT)信号通路的影响。方法采用四甲基偶氮唑盐微量酶反应比色法( MTT法)检测不同浓度人参皂苷 Rg5对 HepG2细胞 24、48和 72 h下的细胞存活率，并计算出半数抑制浓度( IC50)以筛选出合适的作用浓度。 Transwell小室实验检测人参皂苷 Rg5对细胞侵袭和迁移的影响，流式细胞仪检测人参皂苷 Rg5对细胞周期和凋亡的影响，蛋白质印迹法( Western blotting)检测人参皂苷 Rg5对细胞中 AKT、磷酸化蛋白激酶 B(pAKT)、细胞周期蛋白 D1(cyclin D1)、基质金属蛋白酶 9(MMP-9)和 B细胞淋巴瘤 -2(Bcl-2)表达的影响。结果与对照( 0 μmol/L)组相比， 40、80和 160 μmol/L人参皂苷 Rg5 24 h下细胞存活率降低( P<0.05)，20、40、80和 160 μmol/L人参皂苷 Rg5 48 h下细胞存活率明显降低( P<0.05)不同浓度的人参皂苷 72 h下均可使细胞存活率降低( P<0.05)。人参皂苷 Rg5作用 HepG2细胞 24、48和 72 h后的 IC50值分别为1，38.60、91.12和 46.92 μmol/L。与对照( 0 μmol/L)组相比， 40、80和 160 μmol/L处理组中侵袭细胞数［(57.38±3.65)个，(35.26±2.15)个，(32.48±2.23)个比( 82.55±6.02)个］、迁移细胞数［(94.25±6.12)个，(51.57±3.15)个，(48.73±3.26)个比( 137.62±8.83)个］、细胞在 S期的百分比［( 30.06±1.75)%，(23.32±1.51)%，(21.95±1.13)%比( 36.85±2.26)%］和 p-AKT［( 0.61±0.04)，(0.32±0.03)，(0.28±0.03)比( 0.79±0.06)］、 cyclin D1、MMP-9、Bcl-2蛋白的表达水平均明显降低( P<0.05)而细胞凋亡率［( 12.58±2.12)%，(28.35±2.26)%，(25.27±3.23)%比( 5.06±1.25)%］、细胞在 G0/G1期的百分比［( 52.85±3.4264.54±4.03)%，(67.02±4.15)%比( 44.16±3.08)%］均明显升高( P<0.05)。结论人参皂苷 Rg5可抑制肝癌 HepG2细胞增殖、侵袭和迁移并诱导细胞周期阻滞和凋亡，其作用机制可能与抑制 AKT信号通路活化有关。 |
| 英文摘要: |
| Objective To investigate the effects of ginsenoside Rg5 on the biological behavior of HepG2 cells and protein kinase B (AKT) signaling pathway.Methods Methyl thiazolyl tetrazolium (MTT) assay was used to detect the cell viability of HepG2 cells at different concentrations of ginsenoside Rg5 at 24, 48 and 72 hours, and half inhibitory concentration (IC50) was calculated to screen the appropriate concentration. Transwell cell assay was used to detect the effects of ginsenoside Rg5 on cell invasion and migration. Flow cytometry was used to detect the effects of ginsenoside Rg5 on cell cycle and apoptosis. Western blotting was used to detect the effects ofginsenoside Rg5 on the expression of AKT, p-AKT, cyclin D1, matrix metalloproteinase (MMP)-9 and B cell lymphoma (Bcl)-2 proteins. Results Compared with the control group (0 μmol/L), the cell survival rate of 40, 80 and 160 μmol/L ginsenoside Rg5 decreased at 24 h (P<0.05), and that of 20, 40, 80 and 160 μmol/L ginsenoside Rg5 decreased at 48 h (P<0.05), and that of different concentrations of ginsenoside Rg5 decreased at 72 h (P<0.05). IC50 values of HepG2 cells treated with ginsenoside Rg5 for 24, 48 and 72 h were138.60, 91.12 and 46.92 μmol/L, respectively. Compared with the control group (0 μmol/L), the number of invasive cells [(57.38±3.65),(35.26±2.15), (32.48±2.23) vs. (82.55±6.02)],the number of migrating cells [(94.25±6.12), (51.57±3.15), (48.73±3.26) vs. (137.62± 8.83)], the percentage of cells in S phase [(30.06±1.75)%, (23.32±1.51)%, (21.95±1.13)% vs. (36.85±2.26)%] and the expression levels of p-AKT [(0.61±0.04), (0.32±0.03), (0.28±0.03) vs. (0.79±0.06)], cyclin D1, MMP-9 and Bcl-2 proteins in 40, 80 and 160 μmol/L treatment groups were significantly decreased (P<0.05), while the apoptotic rate [(12.58±2.12)%, (28.35±2.26)%, (25.27±3.23)% vs. (5.06± 1.25)%] and the percentage of cells in G0/G1 phase [(52.85±3.42)%, (64.54±4.03)%, (67.02±4.15)% vs. (44.16±3.08)%] were significantly increased (P<0.05).Conclusion Ginsenoside Rg5 can inhibit the proliferation, invasion and migration of HepG2 cells and induce cell cycle arrest and apoptosis. Its mechanism may be related to the inhibition of AKT signaling pathway activation. |
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