Zhao Jinyan, Liu Liya, Wan Yun, Zhang Yuchen, Zhuang Qunchuan, Zhong Xiaoyong, Hong Zhenfeng, Peng Jun
Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian Province, China.
Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian Province, China
Integr Cancer Ther. 2015 Jul;14(4):394-401. doi: 10.1177/1534735415583553. Epub 2015 Apr 27.
We evaluated the effects of total alkaloids of Rubus alceifolius Poir (TARAP) on the migration and invasion of hepatocellular carcinoma (HCC) and furthermore investigated the possible molecular mechanisms mediating its anticancer activity.
We implanted nude mice with human HCC HepG2 cells and fed them with vehicle (physiological saline) or 3 g/kg/day dose of TARAP 5 days per week for 21 days. We determined the in vitro effect of TARAP on the migration and invasion of HepG2 cells by transwell assay. We evaluated SHH signaling components' (SHH, PTCH, SMO, and Gli1) expression levels by reverse transcriptase-polymerase chain reaction and immunohistochemistry. Activity of the matrix metalloproteinases (MMPs) in supernatants was analyzed by zymography. The expression of the MMPs and their specific tissue inhibitor (tissue inhibitor of matrix metalloproteinases, TIMP-1, 2) in HCC tissues was detected by immunohistochemistry.
We discovered that TARAP inhibited hepatocellular migration and invasion in a dose-dependent manner in vitro. In addition, TARAP decreased the expression of SHH, PTCH, SMO, and Gli1 in HCC mouse tumors at both transcriptional and translational levels. Moreover, TARAP inhibited the activity of MMP2 and MMP9. We found that TARAP reduced the expression of MMP2 and MMP9, as well as the tissue inhibitor of MMPs.
Our study showed that TARAP inhibits HCC migration and invasion likely through suppression of the hedgehog pathway. This may, in part, explain its anticancer properties. These results suggest that total alkaloids in Rubus alceifolius may have potential as a novel antimetastasis drug in the treatment of HCC.
我们评估了粗叶悬钩子总生物碱（TARAP）对肝细胞癌（HCC）迁移和侵袭的影响，并进一步研究了介导其抗癌活性的可能分子机制。
我们将人肝癌HepG2细胞接种到裸鼠体内，每周5天给它们喂食溶剂（生理盐水）或3 g/kg/天剂量的TARAP，持续21天。我们通过Transwell实验测定TARAP对HepG2细胞迁移和侵袭的体外作用。我们通过逆转录聚合酶链反应和免疫组织化学评估刺猬信号通路相关蛋白（SHH、PTCH、SMO和Gli1）的表达水平。通过酶谱分析法分析上清液中基质金属蛋白酶（MMPs）的活性。通过免疫组织化学检测HCC组织中MMPs及其特异性组织抑制剂（基质金属蛋白酶组织抑制剂，TIMP-1、2）的表达。
我们发现TARAP在体外以剂量依赖的方式抑制肝细胞的迁移和侵袭。此外，TARAP在转录和翻译水平上均降低了HCC小鼠肿瘤中SHH、PTCH‘ SMO和Gli1的表达。此外，TARAP抑制了MMP2和MMP9的活性。我们发现TARAP降低了MMP2和MMP9以及基质金属蛋白酶组织抑制剂的表达。
我们的研究表明，TARAP可能通过抑制刺猬信号通路来抑制HCC的迁移和侵袭。这可能部分解释了其抗癌特性。这些结果表明，粗叶悬钩子中的总生物碱可能具有作为治疗HCC的新型抗转移药物的潜力。
