Ethyl acetate extract from Jiedu Xiaozheng Yin inhibits the proliferation of human hepatocellular carcinoma cells by suppressing polycomb gene product Bmi1 and Wnt/β-catenin signaling
Ethyl acetate extract from Jiedu Xiaozheng Yin inhibits the proliferation of human hepatocellular carcinoma cells by suppressing polycomb gene product Bmi1 and Wnt/β-catenin signaling
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DOI：
10.3892/or.2014.3541
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发表时间：
2014-12-01
期刊：
影响因子：
4.2
通讯作者：
Du, Jian
中科院分区：
文献类型：
--
作者：
Chen, Xu-Zheng;Cao, Zhi-Yun;Du, Jian
解毒消症饮是一种用于治疗肝癌的中药汤剂。既往研究表明，健心消可通过诱导肝癌细胞G 0/G1期阻滞而抑制肝癌细胞增殖。在这项研究中，我们研究了JXY对HCC细胞的抑制作用是否与Wnt/β-catenin途径和polycomb基因产物Bmi 1的抑制有关。从JXY中制备乙酸乙酯提取物（EE-JXY）。采用MTT法和集落形成实验检测细胞增殖。免疫荧光法检测β-catenin和Bmi 1的表达和定位。免疫组化法检测增殖细胞核抗原（PCNA）、c-myc和cyclin D1的表达。RT-PCR检测β-catenin、Bmi 1、c-myc、cyclin D1和p16（INK 4A）mRNA水平。结果表明，EE-JXY可抑制PCNA、c-myc、cyclin D1和Bmi 1的表达，上调p16（INK 4A）的表达。我们还发现EE-JXY可以促进β-catenin从细胞质和细胞核转位到细胞膜。最后，EE-JXY在HCC的小鼠异种移植模型中证实了细胞增殖和Bmi 1和Wnt/β-连环蛋白表达的抑制。因此，EE-JXY可部分通过抑制Bmi 1和Wnt/beta-catenin信号通路来抑制HCC的增殖。
Jiedu Xiaozheng Yin (JXY) is a Chinese herbal decoction used to treat hepatocellular carcinoma (HCC). Previous studies have demonstrated that JXY can inhibit HCC cell proliferation via induction of G0/G1 phase arrest. In this study, we investigated whether the inhibitory effect of JXY on HCC cells is associated with the inhibition of the Wnt/beta-catenin pathway and the polycomb gene product Bmi1. Ethyl acetate extract from JXY (EE-JXY) was prepared. Methyl thiazolyl tetrazolium (MTT) and colony formation assays were used to measure cell proliferation. Immunofluorescence was used to analyze the expression and location of beta-catenin and Bmi1. Immunohistochemistry was used to examine the expression of proliferating cell nuclear antigen (PCNA), c-myc and cyclin D1. beta-catenin, Bmi1, c-myc, cyclin D1 and p16(INK4A) mRNA levels were detected by RT-PCR. The results demonstrated that EE-JXY inhibited the expression of PCNA, c-myc, cyclin D1 and Bmi1, and upregulated the expression of p16(INK4A). We also found that EE-JXY could facilitate beta-catenin translocation from the cytoplasm and nuclei to the cytomembrane. Finally, suppression of cell proliferation and expression of Bmi1 and Wnt/beta-catenin by EE-JXY was confirmed in a mouse xenograft model of HCC. Thus, EE-JXY can inhibit the proliferation of HCC partially via suppression of the Bmi1 and Wnt/beta-catenin signaling pathways.