Ginsenoside Rh2 and Rh3 induce differentiation of HL-60 cells into granulocytes:: modulation of protein kinase C isoforms during differentiation by ginsenoside Rh2
Ginsenoside Rh2 and Rh3 induce differentiation of HL-60 cells into granulocytes:: modulation of protein kinase C isoforms during differentiation by ginsenoside Rh2
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DOI：
10.1016/s1357-2725(97)00141-6
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发表时间：
1998-03-01
影响因子：
4
通讯作者：
Kim, SI
中科院分区：
文献类型：
--
作者：
Kim, YS;Kim, DS;Kim, SI
人参皂苷Rh-1或Rh-2使B16黑色素瘤或F9畸胎瘤分化为表型正常的黑素细胞样细胞或顶叶内胚层样细胞。人参皂苷Rh-3和Rh-4是最近从人参中分离得到的，但其生化和药理作用尚不清楚。本研究探讨了具有相似结构的人参皂苷Rh基团(G-Rh-1、-Rh-2、-Rh-3和-Rh-4)是否诱导HL-60细胞分化，以及蛋白激酶C(PKC)是否参与人参皂苷的诱导分化作用。Wright-Giemsa染色和硝基蓝四氮唑还原分析细胞分化程度。G-Rh-2和G-Rh-3诱导HL-60细胞分化为形态和功能粒细胞，而G-Rh-1和G-Rh-4不能。G-Rh-2和G-Rh-3使细胞周期停滞在G1/S期，这与维甲酸>；G-Rh-2>；G-Rh-3诱导分化的能力一致。在G-Rh-2诱导分化过程中，胞浆和细胞总抽提液中钙/磷脂依赖的蛋白激酶C活性增加，38和200 kDa内源蛋白的钙/磷脂依赖性磷酸化增加，而60、、66和97 kDa蛋白的磷酸化不依赖于钙/磷脂。免疫印迹分析胞浆PKC亚型时，α水平没有明显变化，但G-Rh-2处理后出现了与PKC催化片段质量相似的60 kDa免疫反应条带。随着治疗时间的延长，β亚型逐渐增加。未处理的细胞胞浆中未检测到伽马异构体，处理5天后检测到少量。结论：G-Rh-2和G-Rh-3可诱导HL-60细胞向粒细胞分化，PKC亚型水平的调节可能是G-Rh-2诱导HL-60细胞分化的机制之一。(C)1998爱思唯尔科学有限公司。保留所有权利。
Ginsenoside Rh-1 or Rh-2 differentiated B16 melanoma or F9 teratocarnoma to phenotypic normal melanocyte-like cells or parietal endoderm-like cells. Ginsenoside Rh-3 and Rh-4 were recently isolated from Panax ginseng, but their biochemical and pharmacological effects remain unidentified. The present study investigated whether the ginsenoside Rh group (G-Rh-1, -Rh-2, -Rh-3 and -Rh-4) having similar structures induce differentiation of HL-60 cells and whether protein kinase C (PKC) is involved in differentiation by ginsenoside. Differentiation was assessed by Wright-Giemsa stain and nitroblue tetrazolium reduction. G-Rh-2 and G-Rh-3 induced differentiation of HL-60 cells into morphologically and functionally granulocytes but G-Rh-1 and G-Rh-4 did not. G-Rh-2 and G-Rh-3 arrested the cell cycle at the G1/S phase, consistent with the ability to induce differentiation in a decreasing order of retinoic acid > G-Rh-2 > G-Rh-3. During differentiation by G-Rh-2, Ca2+/phospholipid-dependent PKC activity was increased in both the cytosol and total cell extract and Ca2+/phospholipid-dependent phosphorylation of 38 and 200 kDa endogenous proteins increased, while phosphorylation of 60, 64, 66 and 97 kDa proteins was Ca2+/phospholipid-independent. When cytosolic PKC isoforms were analyzed by immunoblotting, no significant change was observed in the alpha level, however, the immunoreactive 60 kDa band of a similar mass to the PKC catalytic fragment appeared following treatment with G-Rh-2. The beta isoform was gradually increased with prolonged treatment. The gamma isoform was not detected in the cytosol of untreated cells, whereas a small amount was detected 5 days after treatment. It is concluded that G-Rh-2 and G-Rh-3 can induce differentiation of HL-60 cells into granulocytes and modulation of PKC isoform levels may contribute to differentiation of HL-60 cells by G-Rh-2. (C) 1998 Elsevier Science Ltd. All rights reserved.