Norcantharidin Inhibits SK-N-SH Neuroblastoma Cell Growth by Induction of Autophagy and Apoptosis

Abstract

Norcantharidin, a low-toxic analog of the active anticancer compound cantharidin in Mylabris, can inhibit proliferation and induce apoptosis of multiple types of cancer cells. However, the anticancer activities of norcantharidin with respect to neuroblastoma, and its underlying mechanisms, have not been investigated. Therefore, our study was designed to determine the efficacy of norcantharidin on SK-N-SH neuroblastoma cell death and to elucidate detailed mechanisms of activity. In the present study, norcantharidin suppressed the proliferation and cloning ability of SK-N-SH cells in a dose-dependent manner, apparently by reducing the mitochondrial membrane potential and arresting SK-N-SH cells at the G2/M stage, accompanied by elevated expressions of p21 and decreased expressions of cyclin B1 and cell division control 2. Treatment by norcantharidin induced significant mitophagy and autophagy, as demonstrated by a decrease in Translocase Of Outer Mitochondrial Membrane 20 (TOM20), increased beclin1 and LC3-II protein expression, reduced protein SQSTM1/p62 expression, and accumulation of punctate LC3 in the cytoplasm of SK-N-SH cells. In addition, norcantharidin induced apoptosis through regulating the expression of B-cell lymphoma 2-associated X protein/B-cell lymphoma 2 and B-cell lymphoma 2-associated X protein/myeloid cell leukemia 1 and activating caspase-3 and caspase-9-dependent endogenous mitochondrial pathways. We also observed an increase in phosphor-AMP-activated protein kinase accompanied with a decrease in phosphor-protein kinase B and mammalian target of rapamycin expression after treatment with norcantharidin. Subsequent studies indicated that norcantharidin participates in cellular autophagy and apoptosis via activation of the c-Jun NH2-terminal kinases/c-Jun pathway. In conclusion, our results demonstrate that norcantharidin can reduce the mitochondrial membrane potential, induce mitophagy, and subsequently arouse cellular autophagy and apoptosis; the AMP-activated protein kinase, protein kinase B/mammalian target of rapamycin, and c-Jun NH2-terminal kinases/c-Jun signaling pathways are widely involved in these processes. Thus, the traditional Chinese medicine norcantharidin could be a novel therapeutic strategy for treating neuroblastoma.

Downregulation of c‑FLIP and upregulation of DR‑5 by cantharidin sensitizes TRAIL‑mediated apoptosis in prostate cancer cells via autophagy flux

“…certain studies have reported that cantharidin attenuates tumor cell propagation and induction of autophagy (31)(32)(33)(34). Autophagy has been widely investigated in response to various stimuli, including particular anticancer agents (35).…”

Norcantharidin inhibits viability and induces cell cycle arrest and apoptosis in osteosarcoma

“…NCTD, the demethyl-. NCTD, the demethyl-the demethylated form of cantharidin, has exhibited anticancer potential in multiple cancer cell types (8,12,13). However, to the best of our knowledge, the ability of NCTD to suppress the growth of human osteosarcoma cells and the possible underlying mecha-osteosarcoma cells and the possible underlying mecha-and the possible underlying mechanisms have never been investigated.…”

The Inhibitory Effect of Carboplatin Injection on Human Neuroblastoma SK-N-SH

“…SK-N-SH cells were purchased from the Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (Shanghai, China). Dulbecco's modified eagle medium (DMEM, 12100-46; Thermo Fisher Scientific, MA, USA) containing 10% fetal bovine serum was cultured in an incubator at 37°C with 5% CO 2 20 – 22 . The specification of carboplatin injection is 10 mL:100 mg (Qilu Pharmaceutical Co., Ltd, Jinan City, Shandong Province; batch no.…”