大蒜素通过SHP-1介导的STAT3信号通路抑制胆管癌的体外和体内增殖及侵袭。

Allicin Inhibits Proliferation and Invasion in Vitro and in Vivo via SHP-1-Mediated STAT3 Signaling in Cholangiocarcinoma.

作者信息

Chen Huinan, Zhu Biqiang, Zhao Lei, Liu Yang, Zhao Fuya, Feng Jing, Jin Ye, Sun Jiayu, Geng Rui, Wei Yunwei

机构信息

Department of Oncological and Endoscopic Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China.

Translational Medicine Research and Cooperation Center of Northern China, Harbin, China.

出版信息

Cell Physiol Biochem. 2018;47(2):641-653. doi: 10.1159/000490019. Epub 2018 May 22.

DOI:10.1159/000490019

PMID:29794468

Abstract

BACKGROUND/AIMS: Cholangiocarcinoma (CCA) is a malignant tumor that is resistant to chemotherapy, so new therapeutic agents are needed. Allicin which is rapidly converted from allin by allinase, is one of the most biologically active compounds in freshly crushed garlic and has been shown to have strong anti-tumor effects. Our aim was to explore the molecular mechanism by which allicin affects the cell proliferation and invasion of CCA.

METHODS

Cell viability and apoptosis were measured using the CCK-8 assay, colony formation assay, and flow cytometry. Cell migration and invasion were evaluated by wound healing and Transwell assays, respectively. The expression of several proteins involved in cell apoptosis and invasion were assessed by Western blot. The activation of STAT3 signaling was detected by Western blot and immunofluorescence staining. The involvement of SHP-1 was determined using small interfering RNA (siRNA). Moreover, a nude mouse model of human CCA was established to assess the anti-tumor effects of allicin in vivo.

RESULTS

Allicin significantly suppressed CCA cell proliferation by activating the caspase cascade, inducing apoptosis, and reducing the expression of proteins downstream of STAT3, such as B-cell lymphoma 2 (Bcl-2), while upregulating Bcl-2-associated X (Bax) protein. In addition, allicin inhibited the migration, invasion, and epithelial-mesenchymal transition (EMT) of CCA cells. Moreover, the protein expression of MMP-2 and MMP-9 was significantly downregulated in CCA cells treated with allicin compared with CCA cells treated with control. Mechanistic investigations indicated that allicin upregulated SHP-1 expression in CCA, and pervanadate treatment reversed the allicin-induced downregulation of STAT3. Moreover, suppression of SHP-1 by siRNA overturned the effect of allicin on the induction of SHP-1 and inhibition of STAT3 activation. Additionally, treatment with allicin attenuated tumor growth in the nude mouse model of CCA.

CONCLUSIONS

Our findings suggest that allicin suppresses cell proliferation and invasion via STAT3 signaling and may be a potential therapeutic agent for CCA.

摘要

背景/目的：胆管癌（CCA）是一种对化疗耐药的恶性肿瘤，因此需要新的治疗药物。蒜素由蒜氨酸通过蒜氨酸酶迅速转化而来，是新鲜压碎大蒜中生物活性最强的化合物之一，已被证明具有强大的抗肿瘤作用。我们的目的是探讨蒜素影响CCA细胞增殖和侵袭的分子机制。

方法

使用CCK-8法、集落形成试验和流式细胞术检测细胞活力和凋亡。分别通过伤口愈合试验和Transwell试验评估细胞迁移和侵袭。通过蛋白质印迹法评估参与细胞凋亡和侵袭的几种蛋白质的表达。通过蛋白质印迹法和免疫荧光染色检测STAT3信号的激活。使用小干扰RNA（siRNA）确定SHP-1的参与情况。此外，建立人CCA裸鼠模型以评估蒜素在体内的抗肿瘤作用。

结果

蒜素通过激活半胱天冬酶级联反应、诱导凋亡和降低STAT3下游蛋白（如B细胞淋巴瘤2（Bcl-2））的表达，同时上调Bcl-2相关X蛋白（Bax），显著抑制CCA细胞增殖。此外，蒜素抑制CCA细胞的迁移、侵袭和上皮-间质转化（EMT）。此外，与对照组处理的CCA细胞相比，用蒜素处理的CCA细胞中MMP-2和MMP-9的蛋白表达显著下调。机制研究表明，蒜素上调CCA中SHP-1的表达，过钒酸盐处理逆转了蒜素诱导的STAT3下调。此外，siRNA抑制SHP-1推翻了蒜素对SHP-1诱导和STAT3激活抑制的作用。此外，蒜素处理减弱了CCA裸鼠模型中的肿瘤生长。