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阿尔特罗诺诱导黑色素瘤b16-f0细胞分化。

Alteronol induces differentiation of melanoma b16-f0 cells.

作者信息

Wang Caixia, Zhang Bo, Chen Na, Liu Liangliang, Liu Jinglei, Wang Qi, Wang Zhenhua, Sun Xiling, Zheng Qiusheng

机构信息

Binzhou Medical University, Yantai 264003, P.R. China.

出版信息

Recent Pat Anticancer Drug Discov. 2015;10(1):116-27. doi: 10.2174/1574892809666140923125521.

Abstract

Alteronol, isolated from microbial mutation strains, has been applied for Chinese and International patents for tumor treatment. The aim of this project study is to investigate characteristics of proliferation and redifferentiation induced by alteronol in B16-F0 mouse melanoma cells. Cell proliferation is determined by tetrazolium salt colorimetric method (MTT assay). Morphological changes were analyzed by using Giemsa staining. The levels of melanin and tyrosinase were measured by spectrophotometry. The mRNA expressions of tyrosinase-related protein Trp1 and Trp2 were evaluated by reverse transcription-polymerase chain reaction (RT-PCR). The anchorage-independent proliferation of B16-F0 was monitored by the colony formation assay. Tumorigenicity was characterized by an animal model in vivo. The results showed that the proliferation of B16-F0 cells was inhibited by alteronol in a concentration and time dependent manner. All well-known evaluation indexes of melanoma cell differentiation, including morphological changes and tyrosinase activity alteration, were greatly enhanced with the increase of alteronol concentrations. Taken together, the expression of tyrosinase related gene, decreased cell colony formation rate and the tumorigenicity in vivo; all of these revealed that alteronol plays a key role in inducing differentiation and suppressing the proliferation of B16-F0 tumor cells in vitro and in vivo.

摘要

从微生物突变菌株中分离得到的阿替罗醇已在中国和国际上申请了肿瘤治疗专利。本项目研究的目的是探讨阿替罗醇诱导B16-F0小鼠黑色素瘤细胞增殖和再分化的特性。采用四氮唑盐比色法(MTT法)测定细胞增殖。用吉姆萨染色分析形态学变化。用分光光度法测定黑色素和酪氨酸酶水平。通过逆转录聚合酶链反应(RT-PCR)评估酪氨酸酶相关蛋白Trp1和Trp2的mRNA表达。通过集落形成试验监测B16-F0的非贴壁依赖性增殖。通过体内动物模型表征致瘤性。结果表明,阿替罗醇以浓度和时间依赖性方式抑制B16-F0细胞的增殖。随着阿替罗醇浓度的增加,黑色素瘤细胞分化的所有知名评估指标,包括形态学变化和酪氨酸酶活性改变,均显著增强。综上所述,酪氨酸酶相关基因的表达、细胞集落形成率降低以及体内致瘤性降低;所有这些都表明阿替罗醇在体外和体内诱导B16-F0肿瘤细胞分化和抑制增殖中起关键作用。

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