ABSTRACT
Hepatocellular carcinoma progression is thought to be driven by cancer stem cells (CSCs). No clinical trial has, as yet, shown convincing long-term disease free survival results for the majority of patients in HCC. So it is important to discover new anti-cancer agents. In our study, we chose sophocarpine, which is derived from the foxtail-like sophora herb, for its efficacy to inhibit HCC including CSCs and potential mechanism study. Our results show that sophocarpine could not only reduce HCC cell viability, eliminate HCC and reverse hepatoma cells malignant phenotype, but also reduce the ratio of CSCs and inhibit the sphere formation of CSCs in vitro. In vivo, sophocarpine significantly displayed antitumor effects in subcutaneous xenograft HCC models and orthotopic transplantation tumor models. Further studies showed that sophocarpine could exert anti-tumor effects partly via downregulating the activity of the cancer stem cell related pathways and inhibiting EMT induced by TGF-ß.
Subject(s)
Alkaloids/pharmacology , Antineoplastic Agents, Phytogenic/pharmacology , Carcinoma, Hepatocellular/drug therapy , Catenins/metabolism , Cell Differentiation/drug effects , Epithelial-Mesenchymal Transition/drug effects , Glycogen Synthase Kinase 3 beta/metabolism , Liver Neoplasms/drug therapy , Neoplastic Stem Cells/drug effects , Proto-Oncogene Proteins c-akt/metabolism , Transforming Growth Factor beta/metabolism , Animals , Carcinoma, Hepatocellular/enzymology , Carcinoma, Hepatocellular/pathology , Cell Line, Tumor , Cell Survival/drug effects , Dose-Response Relationship, Drug , Humans , Liver Neoplasms/enzymology , Liver Neoplasms/pathology , Male , Mice, Inbred BALB C , Mice, Nude , Neoplastic Stem Cells/enzymology , Neoplastic Stem Cells/pathology , Phenotype , Signal Transduction/drug effects , Spheroids, Cellular , Time Factors , Tumor Burden/drug effects , Xenograft Model Antitumor AssaysABSTRACT
OBJECTIVE: To explore the effects of total flavonoids of Hippophae rhamnoides L. (TFH), quercetin (Que) and isorhamnetin (Isor) on the intracellular free calcium ([Ca(2+)](i)) in vascular smooth muscle cells (VSMC) of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY). METHODS: Fluo 3-acetoxymethylester (Fluo-3/AM) was used to observe the effects of TFH (100 mg/L) and its essential monomers, namely Que (10(-4) mol/L) and Isor (10(-4) mol/L) on changes of [Ca(2+)](i) in cultured SHR and WKY VSMC (abbr. to Ca-SHR & Ca-WKY) following exposure to high K(+), norepinephrine (NE) and angiotensin II (Ang II), and to compare with the effects of verapamil (Ver). RESULTS: (1) TFH, Que and Isor had inhibitory effects on resting Ca-SHR (P < 0.05), but had no significant effects on Ca-WKY (P > 0.05). (2) High K(+) could increase Ca-SHR more significantly than Ca-WKY (P < 0.05); TFH, Que and Isor could inhibit the elevation of [Ca(2+)](i) induced by high K(+)-depolarization, with the effects similar to that of Ver, and the effect on Ca-SHR was more significant than that on Ca-WKY (P < 0.05). (3) NE and Ang II could increase Ca-SHR more significantly than Ca-WKY (P < 0.05), TFH, Que and Isor had remarkably inhibitory effect on the elevation of Ca-SHR and Ca-WKY induced by NE or Ang II. (4) In the absence of extracellular Ca(2+), TFH, Que and Isor also had certain inhibitory effect on Ca-SHR and Ca-WKY induced by NE, and the effect on the former was more significant than that on the latter (P < 0.05). CONCLUSION: TFH, Que and Isor might decrease the levels of [Ca(2+)](i) in VSMCs by blocking both voltage-dependent calcium channels (VDC) and receptor-operated calcium channels (ROC) in physiological or pathological state, which may be one of the important mechanisms of their hypotensive and protective effects on target organs in patients with hypertension.
