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烯酰辅酶A水合酶短链1、表皮生长因子及低糖环境促进肝癌细胞侵袭转移
ECHS1, epidermal growth factor, and low-glucose conditions promote the invasion and metastasis of hepatocellular carcinoma cells
文章发布日期:2018年06月07日  来源:  作者:卢嘉臻,徐文娟,蔡艺玲,等  点击次数:154次  下载次数:12次

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【摘要】:目的 研究肝细胞癌(HCC)组织中能量代谢相关酶的表达及对肝癌细胞侵袭能力的影响,探讨烯酰辅酶A水合酶短链1(ECHS1)、AMP蛋白激酶(AMPK)、脂肪合成酶(FAS)、乙酰CoA羧化酶(ACC)等相关酶及低糖环境在HCC发生发展过程中的作用。方法 选取解放军第一七五医院2015年6月-2016年6月经手术切除的32例HCC患者肝癌及癌旁组织,通过免疫组化技术,检测不同分化程度肝癌及癌旁组织中能量代谢过程中关键酶FAS、ACC及AMPK等表达情况。构建两组肝癌细胞株,对照组:转染空白对照质粒的肝癌细胞Huh7-plv、HepG2-PU6;实验组:转染小干扰RNA(siRNA)的肝癌细胞株Huh7-siECHS1、HepG2-siECHS1。应用Transwell小室实验观察ECHS1及低糖培养和加入表皮生长因子(EGF)等不同营养条件对肝癌细胞侵袭能力的影响。计量资料组间比较采用t检验;计数资料组间比较采用χ2检验。结果 免疫组化结果显示,肝癌细胞中ACC、AMPKβ表达高于癌旁组织,差异均有统计学意义(21/11 vs 7/25,χ2=12.44,P<0.05;31/1 vs 26/6,χ2=4.68,P<0.05)。Transwell小室实验结果显示,在正常营养条件(Huh7-plv-N、HepG2-PU6-N)及加入EGF的正常营养条件(Huh7-plv-N-EGF、HepG2-PU6-N-EGF)的肝癌细胞侵袭数均明显大于相应干扰ECHS1后(Huh7-siECHS1-N、HepG2-siECHS1-N、Huh7-siECHS1-N-EGF、HepG2-siECHS1-N-EGF)的侵袭细胞数(t值分别为6.93、6.51、7.55、4.93,P值均<0.05);低糖培养环境(Huh7-plv-LowGlu、HepG2-PU6-LowGlu)及加入EGF的低糖培养环境(Huh7-plv-LowGlu-EGF、HepG2-PU6-LowGlu-EGF)的肝癌细胞侵袭数亦大于相应干扰ECHS1后(Huh7-siECHS1-LowGlu、HepG2-siECHS1-LowGlu、Huh7-siECHS1-LowGlu-EGF、HepG2-siECHS1-LowGlu-EGF)的侵袭细胞数,差异均有统计学意义(t值分别为9.52、5.80、20.52、8.80,P值均<0.05)。表明不同营养条件下,干扰ECHS1后肝癌细胞Huh7、HepG2的侵袭能力均明显降低。此外,加入EGF后(Huh7-plv-N-EGF、HepG2-PU6-N-EGF)与正常营养条件(Huh7-plv-N、HepG2-PU6-N)相比,穿透小室的细胞数明显增多(t值分别为4.44、3.17,P值均<0.05);而低糖条件下(Huh7-plv-LowGlu、HepG2-PU6-LowGlu)与正常营养环境相比,发生侵袭的细胞数有所增加,但差异均无统计学意义(P值均>0.05)。结论 ECHS1、EGF可明显增强肝癌细胞侵袭转移能力,低糖饥饿条件亦可对肝癌细胞侵袭能力起到一定促进作用。
【Abstract】:Objective To investigate the expression of key enzymes associated with energy metabolism in hepatocellular carcinoma (HCC) tissue, as well as the roles of short-chain enoyl-CoA hydratase 1 (ECHS1), AMP-activated protein kinase (AMPK), fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), and low-glucose conditions in the development and progression of HCC. Methods A total of 32 HCC patients who underwent surgical resection in 175 Hospital of PLA from June 2015 to June 2016 were enrolled. HCC tissue and adjacent tissue samples were collected, and immunohistochemistry was used to measure the expression of key enzymes associated with energy metabolism in HCC tissue samples with different degrees of tumor differentiation and adjacent tissues, including FAS, ACC, and AMPK. Two groups of hepatoma cells were established. Hepatoma Huh7-plv and HepG2-PU6 cells transfected with blank control plasmids were established as control group, and hepatoma cells transfected with small interfering RNA (siRNA) (Huh7-siECHS1 and HepG2-siECHS1 cells) were established as experimental group. A Transwell chamber assay was used to observe the effect of ECHS1, low-glucose condition, and EGF on the invasion ability of hepatoma cells. The t-test was used for comparison of continuous data between groups, and the chi-square test was used for comparison of categorical data between groups. Results Immunohistochemistry showed that compared with the adjacent tissue samples, the HCC tissue samples had a significant higher proportion of ACC- or AMPKβ-positive cells (ACC-positive cells: 21/11 vs 7/25, χ2=12.44, P<0.05; AMPKβ-positive cells: 31/1 vs 26/6, χ2=4.68, P<0.05). The Transwell chamber assay showed that compared with the corresponding hepatoma cells treated by ECHS1 interference (Huh7-siECHS1-N, HepG2-siECHS1-N, Huh7-siECHS1-N-EGF, and HepG2-siECHS1-N-EGF), the hepatoma cells cultured under normal conditions (Huh7-plv-N and HepG2-PU6-N) and with the addition of EGF (Huh7-plv-N-EGF and HepG2-PU6-N-EGF) had a significantly higher number of invasive cells (t=6.93, 6.51, 7.55, and 4.93, all P<0.05); compared with the corresponding hepatoma cells treated by ECHS1 interference (Huh7-siECHS1-LowGlu, HepG2-siECHS1-LowGlu, Huh7-siECHS1-LowGlu-EGF, and HepG2-siECHS1-LowGlu-EGF), the hepatoma cells cultured under low-glucose conditions (Huh7-plv-LowGlu and HepG2-PU6-LowGlu) and with the addition of EGF (Huh7-plv-LowGlu-EGF and HepG2-PU6-LowGlu-EGF) had a significantly higher number of invasive cells (t=9.52, 5.80, 20.52, and 8.80, all P<0.05). Under different conditions, hepatoma Huh7 and HepG2 cells had a significant reduction in invasion ability after ECHS1 interference. Compared with the hepatoma cells cultured under normal conditions (Huh7-plv-N and HepG2-PU6-N), the hepatoma cells cultured with the addition of EGF (Huh7-plv-N-EGF and HepG2-PU6-N-EGF) had a significant increase in the number of cells passing through the Transwell chamber (t=4.44 and 3.17, both P<0.05). Compared with the hepatoma cells cultured under normal conditions, the hepatoma cells cultured under low-glucose conditions (Huh7-plv-LowGlu and HepG2-PU6-LowGlu) had an increase in the number of invasive cells (both P>0.05). Conclusion ECHS1 and EGF can significantly enhance the invasion and metastasis of hepatoma cells, and low-glucose conditions can also promote the invasion of hepatoma cells.
【关键字】:癌,肝细胞;烯酰CoA水合酶;表皮生长因子;能量代谢;肿瘤侵润;肿瘤转移
【Key words】:carcinoma, hepatocellular; enoyl-CoA hydratase; epidermal growth factor; energy metabolism; neoplasm invasiveness; neoplasm metastasis
【引证本文】:LU JZ, XU WJ, CAI YL, et al. ECHS1, epidermal growth factor, and low-glucose conditions promote the invasion and metastasis of hepatocellular carcinoma cells[J]. J Clin Hepatol, 2018, 34(7): 1481-1486. (in Chinese)
卢嘉臻, 徐文娟, 蔡艺玲, 等. 烯酰辅酶A水合酶短链1、表皮生长因子及低糖环境促进肝癌细胞侵袭转移[J]. 临床肝胆病杂志, 2018, 34(7): 1481-1486.

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