Sann-Joong-Kuey-Jian-Tang induces autophagy in HepG2 cells via regulation of the phosphoinositide-3 kinase/Akt/mammalian target of rapamycin and p38 mitogen-activated protein kinase pathways.
Sann-Joong-Kuey-Jian-Tang induces autophagy in HepG2 cells via regulation of the phosphoinositide-3 kinase/Akt/mammalian target of rapamycin and p38 mitogen-activated protein kinase pathways.
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DOI：
10.3892/mmr.2015.3573
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发表时间：
2015-08
影响因子：
3.4
通讯作者：
Chen YL
中科院分区：
文献类型：
--
作者：
Chuang WL;Su CC;Lin PY;Lin CC;Chen YL
三重归健汤（Sann-Joong-Kuey-Jian-Tang，SJKJT）是一种传统中药，它能通过外源性途径诱导人肝癌细胞系HepG 2的自噬并抑制其增殖。在本研究中，在HepG 2细胞中研究了SJKJT诱导的自噬作用和介导这些作用的细胞毒性机制。使用3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四唑鎓试验评价SJKJT在HepG 2细胞中的细胞毒性。结果表明：SJKJT的半数抑制浓度在24 h为2.91 mg/ml，48 h为1.64 mg/ml，72 h为1.26 mg/ml。共聚焦荧光显微镜观察结果表明，参肾康煎剂可导致绿色荧光蛋白-LC 3的积累和细胞质空泡化。流式细胞仪分析显示酸性囊泡细胞器的积累。此外，用于确定自噬相关蛋白表达水平的蛋白质印迹分析表明，用SJKJT处理的HepG 2细胞表现出LC 3B-I/LC 3B-II转化，Beclin表达水平增加，Atg-3和Atg-5，p62表达水平降低，磷酸肌醇-3激酶/Akt/雷帕霉素哺乳动物靶点和p38丝裂原信号传导降低，激活蛋白激酶途径。总之，这些发现可能有助于开发用于治疗恶性类型肝癌的新型化疗药物。
Sann-Joong-Kuey-Jian-Tang (SJKJT), a traditional Chinese medicine, was previously reported to induce autophagy and inhibit the proliferation of the human HepG2 hepatocellular carcinoma cell line via an extrinsic pathway. In the present study, the effects of SJKJT-induced autophagy and the cytotoxic mechanisms mediating these effects were investigated in HepG2 cells. The cytotoxicity of SJKJT in the HepG2 cells was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The results demonstrated that the half-maximal inhibitory concentration of SJKJT was 2.91 mg/ml at 24 h, 1.64 mg/ml at 48 h and 1.26 mg/ml at 72 h. The results of confocal fluorescence microscopy indicated that SJKJT resulted in the accumulation of green fluorescent protein-LC3 and vacuolation of the cytoplasm. Flow cytometric analysis revealed the accumulation of acidic vesicular organelles. Furthermore, western blot analysis, used to determine the expression levels of autophagy-associated proteins, demonstrated that the HepG2 cells treated with SJKJT exhibited LC3B-I/LC3B-II conversion, increased expression levels of Beclin, Atg-3 and Atg-5 and reduced expression levels of p62 and decreased signaling of the phosphoinositide-3 kinase/Akt/mammalian target of rapamycin and the p38 mitogen-activated protein kinase pathways. Taken together, these findings may assist in the development of novel chemotherapeutic agents for the treatment of malignant types of liver cancer.
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影响因子：
5.6
作者：
Miao Q;Bi LL;Li X;Miao S;Zhang J;Zhang S;Yang Q;Xie YH;Zhang J;Wang SW
通讯作者：
Wang SW
影响因子：
5.4
作者：
Chien, Su-Yu;Kuo, Shou-Jen;Su, Chin-Cheng
通讯作者：
Su, Chin-Cheng
影响因子：
3.6
作者：
Aoki, Hiroshi;Takada, Yasunari;Kondo, Yasuko
通讯作者：
Kondo, Yasuko
影响因子：
9.7
作者：
Hu, Wei;Chen, Sang-Sang;Zhou, Hui-Jun
通讯作者：
Zhou, Hui-Jun
影响因子：
3.5
作者：
Cui, Qlao;Tashiro, Shin-Ichl;Ikejima, Takashi
通讯作者：
Ikejima, Takashi