肝星状细胞通过影响Ang-1/Tie2轴促进肝癌血管新生

doi:10.3877/cma.j.issn.2095-3232.2019.04.021

中华肝脏外科手术学电子杂志 ›› 2019, Vol. 08 ›› Issue (04) : 370 -374. doi: 10.3877/cma.j.issn.2095-3232.2019.04.021

所属专题：文献；

基础研究

肝星状细胞通过影响Ang-1/Tie2轴促进肝癌血管新生

林洋¹, 曹彦龙¹, 邱红¹, 林楠², 吴建华¹^,^†(

)

^1. 844000　新疆维吾尔自治区喀什地区第一医院肝胆外科
^2. 510630　广州，中山大学附属第三医院肝胆外科

收稿日期:2019-05-10 出版日期:2019-08-10
通信作者: 吴建华
基金资助:
新疆维吾尔自治区自然科学基金(2016D01C019)

Hepatic stellate cells promote angiogenesis of hepatocellular carcinoma by affecting Ang-1/Tie2 axis

Yang Lin¹, Yanlong Cao¹, Hong Qiu¹, Nan Lin², Jianhua Wu¹^,^†()

^1. Department of Hepatobiliary Surgery, the First People’s Hospital of Kashgar Area, Xinjiang Uygur Autonomous Region, Kashgar 844000, China
^2. Department of Hepatobiliary Surgery, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China

Received:2019-05-10 Published:2019-08-10
Corresponding author: Jianhua Wu
About author:
Corresponding author: Wu Jianhua, Email: wujianhuakashi007@126.com

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目的

探讨肝星状细胞（HSC）对肝癌细胞血管新生的影响及相关机制。

方法

胶原酶灌注法成功获取人原代肝癌细胞（HC）、人HSC和人肝窦血管内皮细胞（HVEC）。RT-PCR检测3组细胞内血管新生相关基因表达情况。RT-PCR法和ELISA检测HSC在不同浓度TGF-β激活条件下血管生成素（Ang-1）表达情况。RT-PCR及Western Blot检测HVEC与HSC共培养后Tie2基因和蛋白的表达情况。多组Ang-1和Tie2 mRNA比较采用单因素方差分析和LSD-t检验。

结果

HC、HSC和HVEC中Ang-1 mRNA的相对表达量分别为1.6±0.4、7.6±1.6、0.4±0.1，HSC表达量最高（F=94.6，P<0.05）；Tie2 mRNA的相对表达量分别为0.1、1.3、14.4±2.2，HVEC表达量最高（F=239.4，P<0.05）。随着TGF-β刺激浓度的增大，HSC内α-SMA表达量增高，细胞内的Ang-1 mRNA表达增加；细胞培养液上清具有同样的增加趋势。随着HSC活化程度增加，共培养体系内的HVEC表达Tie2 mRNA能力越强，且HSC诱导Tie2蛋白的表达亦增强。

结论

肝癌微环境中活化态HSC是Ang-1的主要来源，HSC可能通过Ang-1/Tie2轴有效促进肝癌血管新生。

关键词: 癌，肝细胞, 肝星状细胞, 血管生成素1, 新生血管化，病理性

Objective

To investigate the effect and its mechanism of hepatic stellate cells (HSC) on the angiogenesis of liver cancer cells.

Methods

Primary human hepatocellular carcinoma cells (HC), human hematopoietic stem cells (HSC) and human vascular endothelial cells (HVEC) were successfully obtained with collagenase perfusion. The expression levels of angiogenesis-related genes in 3 groups were measured by RT-PCR. The expression levels of angiopoietin-1 (Ang-1) in HSCs activated under different concentrations of TGF-β were measured by RT-PCR and ELISA. The expression levels of Tie2 gene and protein in coculture of HVECs and HSCs were detected by RT-PCR and Western blot. The expression levels of Ang-1 and Tie2 mRNA among different groups were statistically compared by univariate ANOVA and LSD-t test.

Results

The relative expression levels of Ang-1 mRNA in HCs, HSCs and HVECs were 1.6±0.4, 7.6±1.6 and 0.4±0.1, respectively, the expression level in HSCs was the highest (F=94.6, P<0.05). The relative expression levels of Tie2 mRNA were 0.1, 1.3 and 14.4±2.2, respectively, with the highest level in HVECs (F=239.4, P<0.05). With the increasing concentration of TGF-β, the expression of α-SMA and Ang-1 mRNA in HSCs was up-regulated. The same increasing trend was observed in the supernatant of cell culture solution. With the increase of HSC activation, the ability of expressing Tie2 mRNA of HVECs in the co-culture system enhanced, and the ability of inducing Tie2 protein expression of HSCs enhanced, too.

Conclusions

Activated HSC is the main source of Ang-1 in the microenvironment of liver cancer. HSC can effectively promote the angiogenesis of liver cancer probably through the Ang-1/Tie2 axis.

Key words: Carcinoma, hepatocellular, Hepatic stellate cells, Angiopoietin-1, Neovascularization, pathologic

图1 HC、HSC、HVEC细胞中Ang-1、Tie2 mRNA表达情况

图2 不同浓度TGF-β激活HSC后Ang-1 mRNA和蛋白分泌情况

图3 RT-PCR和Western blot检测HSC和HVEC共培养后HVEC中Tie2变化

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