Wang Zhi-Min, Song Nan, Ren Yan-Ling
The First Clinical Institute, Liaoning University of Traditional Chinese Medicine, Shenyang, Liaoning 110847, P.R. China.
Key Laboratory of Ministry of Education for Traditional Chinese Medicine Viscera-State Theory and Applications, Liaoning University of Traditional Chinese Medicine, Shenyang, Liaoning 110847, P.R. China.
Mol Med Rep. 2015 Nov;12(5):6815-20. doi: 10.3892/mmr.2015.4282. Epub 2015 Sep 2.
Several biological properties of icariin have been identified, including its anticancer effect. However, the potential mechanisms underlying the effect of icariin on HepG2 hepatocellular carcinoma cells remain to be elucidated. The aim of the present study was to examine the effects of icariin on the proliferation and cytoskeleton of HepG2 cells. A 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5 diphenyltetrazolium bromide assay was used to assess the antiproliferative effects of icariin and to determine the optimal concentration and treatment schedule of icariin on the HepG2 cells. Cell cycle analysis was performed using fluorescence activated cell sorting, the protein expression of B‑cell lymphoma (Bcl)‑2 was determined using immunohistochemical and western blot analyses, and F‑actin in the cells was examined using confocal microscopy. The chemotherapeutic drug, oxaliplatin, was used as a positive control. The results demonstrated that the optimal concentration of icarrin to produce an antiproliferative effect on HepG2 cells was 10‑5 mol/l, and the optimal treatment duration was 72 h. The icariin group had a significantly higher proportion of cells in the G0/G1 phase, compared with the control group, treated with high glucose Dulbecco's modified Eagles medium with 10% fetal bovine serum (P<0.05). The proportion of HepG2 cells in the S phase was significantly lower in the oxaliplatin (24.19%; P<0.05) and icariin (21.07%; P<0.01) groups, compared with the control group (28.62%). Icariin markedly decreased the expression of Bcl‑2, compared with the control (P<0.01), and disrupted the polymerization of F‑actin filaments in the HepG2 cells. Therefore, the present study demonstrated that, at an optimum concentration of 10‑5 mol/l, icariin inhibited the proliferation of the HepG2 cells, promoted apoptosis by decreasing the expression of Bcl‑2, and disrupted the actin cytoskeleton.
淫羊藿苷的多种生物学特性已被确定，包括其抗癌作用。然而，淫羊藿苷对HepG2肝癌细胞作用的潜在机制仍有待阐明。本研究的目的是检测淫羊藿苷对HepG2细胞增殖和细胞骨架的影响。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法评估淫羊藿苷的抗增殖作用，并确定淫羊藿苷对HepG2细胞的最佳浓度和处理时间。使用荧光激活细胞分选进行细胞周期分析，采用免疫组织化学和蛋白质印迹分析测定B细胞淋巴瘤(Bcl)-2的蛋白表达，并使用共聚焦显微镜检查细胞中的F-肌动蛋白。化疗药物奥沙利铂用作阳性对照。结果表明，对HepG2细胞产生抗增殖作用的淫羊藿苷最佳浓度为10-5mol/L，最佳处理时间为72小时。与用含10%胎牛血清的高糖杜尔贝科改良伊格尔培养基处理的对照组相比，淫羊藿苷组处于G0/G1期的细胞比例显著更高(P<0.05)。与对照组(28.62%)相比，奥沙利铂组(24.19%；P<0.05)和淫羊藿苷组(21.07%；P<0.01)中处于S期的HepG2细胞比例显著更低。与对照组相比，淫羊藿苷显著降低了Bcl-2的表达(P<0.01)，并破坏了HepG2细胞中F-肌动蛋白丝的聚合。因此，本研究表明，在最佳浓度10-5mol/L时，淫羊藿苷抑制HepG2细胞的增殖，通过降低Bcl-2的表达促进细胞凋亡，并破坏肌动蛋白细胞骨架。
