Department of Pharmacy, Nanfang Hospital, Southern Medical University, No. 1838, Guangzhou Boulevard (North), Guangzhou 510515, China; Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China.
Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, China; Laboratory of Anti-inflammatory and Immunomodulatory Pharmacology, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, 510515, China.
Phytomedicine. 2021 Jul;87:153580. doi: 10.1016/j.phymed.2021.153580. Epub 2021 Apr 27.
Hepatocellular carcinoma (HCC) is one of the most prevalent and lethal cancer with high metastasis and recurrence rates. Hypoxia-induced miRNAs and HIF-1α are demonstrated to play essential roles in tumor metastasis. Matrine (CHNO), an alkaloid extracted from Sophora flavescens Aiton, has been used as adjuvant therapy for liver cancer in China. The anti-metastasis effects of matrine on HCC and the underlying mechanisms remain poorly understood.
We aimed to investigate the effects of matrine on metastasis of HCC both in vitro and in vivo, and explored whether miR-199a-5p and HIF-1α are involved in the action of matrine.
MTT method, colony formation, wound healing and matrigel transwell assays were performed to evaluate the effects of matrine on cell proliferation, migration and invasion. Nude mice xenograft model and immunohistochemistry (IHC) assay were employed to investigate the anti-metastatic action of matrine in vivo. Quantitative real-time PCR, western blot and dual luciferase reporter assay were conducted to determine the underlying mechanisms of matrine.
Matrine exerted stronger anti-proliferative action on Bel7402 and SMMC-7721 cells under hypoxia than that in normoxia. Both matrine and miR-199a-5p exhibited significant inhibitory effects on migration, invasion and EMT in Bel7402 and SMMC-7721 cells under hypoxia. Further study showed that miR-199a-5p was downregulated in HCC cell lines, and this microRNA was identified to directly target HIF-1α, resulting in decreased HIF-1α expression. Matrine induced miR-199a-5p expression, decreased HIF-1α expression and inhibited metastasis of Bel7402 and SMMC-7721 cells, while miR-199a-5p knockdown reversed the inhibitory effects of matrine on cell migration, invasion, EMT and HIF-1α expression. In vivo, matrine showed significant anti-metastatic activity in the nude mouse xenograft model. H&E and IHC analysis indicated that lung and liver metastasis nodules were reduced, and the protein expression of HIF-1α and Vimentin were significantly decreased by i.p injection of matrine.
Matrine exhibits significant anti-metastatic effect on HCC, which is attributed to enhanced miR-199a-5p expression and subsequently impaired HIF-1α signaling and EMT. These findings suggest that miR-199a-5p is a potential therapeutic target of HCC, and matrine may represent a promising anti-metastatic medication for HCC therapy.
肝细胞癌（HCC）是最常见和最致命的癌症之一，具有高转移和复发率。缺氧诱导的 miRNAs 和 HIF-1α 已被证明在肿瘤转移中发挥重要作用。苦参碱（CHNO），一种从苦参中提取的生物碱，已在中国被用作肝癌的辅助治疗药物。苦参碱对 HCC 的抗转移作用及其潜在机制仍知之甚少。
我们旨在研究苦参碱在体外和体内对 HCC 转移的影响，并探讨 miR-199a-5p 和 HIF-1α 是否参与了苦参碱的作用。
采用 MTT 法、集落形成、划痕愈合和基质胶 Transwell 检测，评估苦参碱对细胞增殖、迁移和侵袭的影响。裸鼠异种移植模型和免疫组织化学（IHC）检测用于研究苦参碱在体内的抗转移作用。实时定量 PCR、Western blot 和双荧光素酶报告基因检测用于确定苦参碱的潜在机制。
苦参碱在缺氧条件下对 Bel7402 和 SMMC-7721 细胞的增殖抑制作用强于常氧条件。苦参碱和 miR-199a-5p 均显著抑制 Bel7402 和 SMMC-7721 细胞在缺氧条件下的迁移、侵袭和 EMT。进一步研究表明，miR-199a-5p 在 HCC 细胞系中下调，该 miRNA 可直接靶向 HIF-1α，导致 HIF-1α 表达降低。苦参碱诱导 miR-199a-5p 表达，降低 HIF-1α 表达，抑制 Bel7402 和 SMMC-7721 细胞的转移，而 miR-199a-5p 敲低则逆转了苦参碱对细胞迁移、侵袭、EMT 和 HIF-1α 表达的抑制作用。在体内，苦参碱在裸鼠异种移植模型中表现出显著的抗转移活性。H&E 和 IHC 分析表明，苦参碱腹腔注射可减少肺和肝转移结节，降低 HIF-1α 和波形蛋白的蛋白表达。
苦参碱对 HCC 具有显著的抗转移作用，这归因于增强的 miR-199a-5p 表达，随后破坏了 HIF-1α 信号和 EMT。这些发现表明，miR-199a-5p 是 HCC 的一个潜在治疗靶点，苦参碱可能代表 HCC 治疗中一种有前途的抗转移药物。
