Department of Laboratory Medicine and Research Center, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, Shaanxi Province, China.
World J Gastroenterol. 2011 Jun 21;17(23):2848-54. doi: 10.3748/wjg.v17.i23.2848.
To observe the effects of sargentgloryvine stem extracts (SSE) on the hepatoma cell line HepG-2 in vitro and in vivo and determine its mechanisms of action.
Cultured HepG-2 cells treated with SSE were analysed by 3-(4,5-Dimethyl-thiazol-2-yl)-2,5-Diphenyltetrazolium bromide and clone formation assay. The cell cycle and apoptosis analysis were conducted by flow cytometric, TdT-Mediated dUTP Nick End Labeling and acridine orange/ethidium bromide staining methods, and protein expression was examined by both reverse transcriptase-polymerase chain reaction and Western blotting. The pathological changes of the tumor cells were observed by haematoxylin and eosin staining. Tumor growth inhibition and side effects were determined in a xenograft mouse model.
SSE treatment could not only inhibit HepG-2 cell proliferation in a dose- and time-dependent manner but also induce apoptosis and cell cycle arrest at the S phase. The number of colonies formed by SSE-treated tumor cells was fewer than that of the controls (P < 0.05). SSE induced caspase-dependent apoptosis accompanied by a significant decrease in Bcl-xl and Mcl-1 and elevation of Bak expression (P < 0.05). Tumor necrosis factor α in the xenograft tumor tissue and the liver functions of SSE-treated mice showed no significant changes at week 8 compared with the control group (P > 0.05). Systemic administration of SSE could inhibit the HepG-2 xenograft tumor growth with no obvious toxic side effects on normal tissues.
SSE can induce apoptosis of HepG-2 cells in vitro and in vivo through decreasing expression of Bcl-xl and Mcl-1 and increasing expression of Bax.
观察杠柳茎提取物（SSE）在体外和体内对肝癌细胞系 HepG-2 的作用，并探讨其作用机制。
用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐和克隆形成试验分析用 SSE 处理的培养的 HepG-2 细胞。通过流式细胞术、末端转移酶介导的 dUTP 缺口末端标记和吖啶橙/溴化乙锭染色法进行细胞周期和凋亡分析，并通过逆转录-聚合酶链反应和 Western blot 检测蛋白表达。用苏木精和伊红染色观察肿瘤细胞的病理变化。在异种移植小鼠模型中测定肿瘤生长抑制和副作用。
SSE 处理不仅可以剂量和时间依赖性地抑制 HepG-2 细胞增殖，还可以诱导凋亡和细胞周期停滞在 S 期。与对照组相比，SSE 处理的肿瘤细胞形成的菌落数量较少（P < 0.05）。SSE 诱导 caspase 依赖性凋亡，同时 Bcl-xl 和 Mcl-1 显著减少，Bak 表达增加（P < 0.05）。与对照组相比，第 8 周时，异种移植肿瘤组织中的肿瘤坏死因子-α和 SSE 处理小鼠的肝功能无明显变化（P > 0.05）。SSE 全身给药可抑制 HepG-2 异种移植肿瘤生长，对正常组织无明显毒性副作用。
SSE 可通过降低 Bcl-xl 和 Mcl-1 的表达，增加 Bax 的表达，诱导 HepG-2 细胞在体外和体内凋亡。
