窖蛋白-1 敲低通过抑制血管内皮生长因子诱导的血管生成降低 SMMC7721 人肝癌细胞的侵袭性。
Caveolin-1 Knockdown Decreases SMMC7721 Human Hepatocellular Carcinoma Cell Invasiveness by Inhibiting Vascular Endothelial Growth Factor-Induced Angiogenesis.
机构信息
Department of Hepatopancreatobiliary Surgery, The First Affiliated Hospital, Fujian Medical University, Fuzhou 350004, China.
出版信息
Can J Gastroenterol Hepatol. 2020 Jun 27;2020:8880888. doi: 10.1155/2020/8880888. eCollection 2020.
BACKGROUND
Recently, several studies have demonstrated that caveolin-1 overexpression is involved in apoptosis resistance, angiogenesis, and invasiveness in hepatocellular carcinoma (HCC). However, the mechanisms underlying caveolin-1-mediated tumor progression remain unclear. . Lentiviral vectors were used to construct caveolin-1 small interfering RNA- (siRNA-) expressing cells. Secreted VEGF levels in SMMC7721 cells were evaluated by enzyme-linked immunosorbent assay (ELISA). SMMC7721 cell proliferation, cycle, apoptosis, and invasiveness were detected by MTT, flow cytometry, Annexin V-FITC/PI, and invasion assay, respectively. Phospho-eNOS levels in human umbilical vein endothelial cells (HUVECs) cocultured with SMMC7721 cell supernatants were analyzed by Western blot. Capillary-like tubule formation assay was performed to analyze endothelial tubular structure formation in HUVECs treated with supernatants from caveolin-1 siRNA-expressing SMMC7721 cells. SMMC7721 implantation and growth in nude mice were observed. Angiogenesis was analyzed by immunohistochemical angiogenesis assay.
RESULTS
Caveolin-1 siRNA-expressing SMMC7721 cells secreted reduced levels of VEGF. Caveolin-1 RNAi also caused an inhibition of SMMC7721 cell proliferation and cell cycle progression that was accompanied by increased apoptosis. Supernatants from caveolin-1 siRNA-expressing SMMC7721 cells inhibited cell cycle progression and decreased phospho-eNOS levels in HUVECs. Endothelial tubular structure formation in HUVECs treated with supernatants from caveolin-1 siRNA-expressing SMMC7721 cells was considerably reduced. Caveolin-1 siRNA-expressing SMMC7721 cells also showed reduced tumorigenicity and angiogenesis induction .
CONCLUSION
Our results reveal a novel mechanism, whereby caveolin-1 positively regulates human HCC cell invasiveness by coordinating VEGF-induced angiogenesis.
背景
最近，有几项研究表明，窖蛋白-1 过表达与肝癌（HCC）中的凋亡抵抗、血管生成和侵袭性有关。然而，窖蛋白-1 介导的肿瘤进展的机制尚不清楚。 . 慢病毒载体用于构建窖蛋白-1 小干扰 RNA-（siRNA-）表达细胞。通过酶联免疫吸附试验（ELISA）评估 SMMC7721 细胞中分泌的 VEGF 水平。通过 MTT、流式细胞术、Annexin V-FITC/PI 和侵袭试验分别检测 SMMC7721 细胞的增殖、周期、凋亡和侵袭。用人脐静脉内皮细胞（HUVEC）与 SMMC7721 细胞上清液共培养，分析其中磷酸化内皮型一氧化氮合酶（eNOS）水平。通过毛细血管样管形成试验分析经窖蛋白-1 siRNA 表达 SMMC7721 细胞上清液处理的 HUVEC 中内皮管状结构的形成。观察裸鼠中 SMMC7721 的植入和生长情况。通过免疫组化血管生成分析评估血管生成。
结果
窖蛋白-1 siRNA 表达的 SMMC7721 细胞分泌的 VEGF 水平降低。窖蛋白-1 RNAi 还导致 SMMC7721 细胞增殖和细胞周期进展受到抑制，同时细胞凋亡增加。来自窖蛋白-1 siRNA 表达 SMMC7721 细胞的上清液抑制了 HUVEC 中的细胞周期进展，并降低了磷酸化 eNOS 水平。用窖蛋白-1 siRNA 表达 SMMC7721 细胞上清液处理的 HUVEC 中内皮管状结构的形成明显减少。窖蛋白-1 siRNA 表达的 SMMC7721 细胞的肿瘤发生和血管生成诱导能力也降低。
结论
我们的研究结果揭示了一种新的机制，即窖蛋白-1 通过协调 VEGF 诱导的血管生成，正向调节人 HCC 细胞的侵袭性。
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