医学临床研究
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医学临床研究  2021, Vol. 38 Issue (1): 68-71    DOI: 10.3969/j.issn.1671-7171.2021.01.020
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安罗替尼对人肝癌细胞裸鼠移植肿瘤生长及生物钟基因表达的影响*
陈卓颖a, 王燕b, 刘文胜a, 柳湘洁a **
华中科技大学同济医学院附属梨园医院:a.老年医学; b.肿瘤内科,湖北 武汉 430077
Effects of Anlotinib on Tumor Growth and Circadian Gene Expression of Hepatocellular Carcinoma Xenografts in Nude Mice
CHEN Zhuo-ying, WANG Yan, LIU Wen-sheng, et al
Department of Geriatrics, Liyuan Hospital of Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430077,China
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摘要 【目的】探讨安罗替尼对人肝癌细胞裸鼠移植后肿瘤生长情况及相应生物钟基因表达的影响。【方法】选取实验无胸腺裸鼠共36只,取人肝癌HepG2细胞,移植于小鼠左侧腋皮下,建立裸鼠皮下肝癌移植瘤模型。接种完成1周后,随机分为安罗替尼处理组和对照组,每组18只。安罗替尼组给予安罗替尼 50 mg/(kg·d)灌胃,空白对照组给予二甲基亚砜(DMSO)100 μL/20(g·d)灌胃,每日1次,连续2周。在灌胃开始前和灌胃结束后24 h测量肿瘤体积情况。待2周后处死小鼠并剥离肿瘤,采用实时PCR(Real-Time PCR)法检测生物钟基因 Per(Per1、Per2、Per3)、CLOCK、Cry(Cry1、Cry2)、Bmal1、Dec2和Timeless mRNA表达水平,同时运用western blot评估上述基因及蛋白质表达情况。【结果】与对照组相比,安罗替尼组裸鼠移植瘤较对照组生长缓慢,体积趋向于缩小。RT-PCR提示肝癌细胞中生物钟基因在安罗替尼作用下,Per1、Per2的mRNA表达量明显增加(P<0.05),同时下游蛋白质表达量增加,而Dec2的mRNA表达量及下游蛋白质表达则明显受抑制(P<0.05);Cry1的mRNA表达量不变,但相应的下游蛋白质表达受抑制。【结论】安罗替尼能显著调节肝癌细胞中多种生物钟基因表达,调控其相应表达产物生成,可抑制肝癌移植瘤的生长,其作用机制之一可能与调控生物钟基因表达密切相关。
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陈卓颖
王燕
刘文胜
柳湘洁
关键词 肝肿瘤实验性/病理学小鼠肿瘤细胞培养的肿瘤移植    
Abstract:【Objective】To investigate the potential effects of anlotinib on the growth of hepatocellular carcinoma (HCC) xenografts in nude mice,and its influences on the circadian (biological clock) gene expression in HCC tissues. 【Methods】We chose 36 athymic nude mice(half male and half female) for the study. The mice were divided randomly into two groups (the observation group and the control group), which were 18 mice per group. HepG2 cells were transplanted under the left axilla of the mice to establish the subcutaneous hepatocellular carcinoma xenograft model . After one week of HepG2 cell implatation, the observation group mice received anlotinib 50mg/(kg·d) for two weeks by intragastric administration, while the control group mice were given vehicle DMSO 100μL/20(g·d) in the same way gavage. Tumor volume was measured before and 24h after gavage. It was measured in every 2 days until experiment ended. In the end, the mice were sacrificed and tumors were isolated. Real-time PCR (rt-PCR) method was used to detect the mRNA level of circadian clock genes Per (Per1, Per2, Per3), CLOCK, Cry (Cry1, Cry2), Bmal1, and Dec2. At the same time we used western blot to evaluate the proteins of circadian genes expression. 【Results】 Compared to the control group, the xenograft tumors of the nude mice in the Anlotinib group grew slowly and tended to shrink in size, which were statistically significant (P<0.05). The results of rt-PCR indicated that the mRNA expressions of Per1 and Per2 in HCC cells were significantly increased under the action of anlotinib (P<0.05), while the downstream proteins increased by western blot. Meanwhile, the mRNA level of Dec2 were significantly inhibited (P<0.05) which accompanied with its downstream protein decreased. Though the mRNA level of Cry1 remained unchanged, however, the downstream protein expression of Cry1 was inhibited. 【Conclusion】Anlotinib can significantly regulate the expressions of various biological clock genes in HCC cells, which thereafter regulate their corresponding protein expression. Thus, anlotinib can significantly inhibit the tumor growth of HCC xenografts.
Key wordsLiver Neoplasms    Experimental/PA    Mice    Nude    Tumor Cells    Cultured    Neoplasm Transplantation
收稿日期: 2019-10-28     
中图分类号:  R735.7  
通讯作者: **E-mail:2019LY0907@hust.edu.cn   
引用本文:   
陈卓颖, 王燕, 刘文胜, 柳湘洁. 安罗替尼对人肝癌细胞裸鼠移植肿瘤生长及生物钟基因表达的影响*[J]. 医学临床研究, 2021, 38(1): 68-71.
CHEN Zhuo-ying, WANG Yan, LIU Wen-sheng, et al. Effects of Anlotinib on Tumor Growth and Circadian Gene Expression of Hepatocellular Carcinoma Xenografts in Nude Mice. JOURNAL OF CLINICAL RESEARCH, 2021, 38(1): 68-71.
链接本文:  
http://journal07.magtech.org.cn/yxlcyj/CN/10.3969/j.issn.1671-7171.2021.01.020     或     http://journal07.magtech.org.cn/yxlcyj/CN/Y2021/V38/I1/68
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