Neferine induces mitochondrial dysfunction to exert anti-proliferative and anti-invasive activities on retinoblastoma

Abstract

Retinoblastoma is common primary intraocular malignancy of infants and childhood. Neferine is a major bisbenzylisoquinoline alkaloid derived from the lotus plumule in Nelumbo nucifera. This study evaluated the mitigation role of Neferine on retinoblastoma in vitro and in vivo. Xenotransplantation model was established by injecting WERI-Rb-1 cells subcutaneously. Upon induction of retinoblastoma , mice were intraperitoneally injected with Neferine (0, 0.5, 1, 2 mg/kg) or ethanol every 3 days for 30 days. Tumor weight and tumor volume were measured every three days and compared between four groups. Then, mice were sacrificed and immunohistochemical examination was performed to compare Ki67, VEGF content between groups. WERI-Rb-1 cells were used for in vitro experiments and the anti-angiogenic role of Neferine was assessed by analyzing nodes/HPF number. In WERI-Rb-1 xenotransplantation model, compared with control group, 1 mg/kg Neferine treatment significantly inhibited tumor weight (0.39 ± 0.04 g vs. 0.25 ± 0.03 g, P< 0.05) and tumor volume (2163 ± 165 mm3 vs. 1276 ± 108 mm3, P< 0.05) after 30 days. Compared with ethanol-injected mice, 2 μM Neferine treatment significantly enhanced apoptosis rate (2.1 ± 0.6% vs. 14.6 ± 2.6%, P< 0.05), accompany downregulation of Ki67 (0.09 ± 0.02% vs. 0.01 ± 0.004%, P< 0.05) and VEGF (0.28 ± 0.04% vs. 0.05 ± 0.03%, P< 0.05) expression. Additionally, 2 μM Neferine treatment significantly decreased JC-1 red/green percentage. High-dose Neferine could decrease retinoblastoma angiogenesis in association with a significant inhibition on tumor growth and invasion. These findings suggested that Neferine could be a new treatment or adjuvant against retinoblastoma.

Lotus (Nelumbo nucifera Gaertn.) and Its Bioactive Phytocompounds: A Tribute to Cancer Prevention and Intervention

“…There is at least one report of the N. nucifera alkaloid neferine on retinoblastoma, a common primary intraocular childhood and infant malignancy. Wang et al [100] observed that neferine inhibited the proliferation and viability of the WERI-Rb-1 human retinoblastoma cell line associated with reduced protein expression of Ki-67 and Survivin. Neferine treatment reduced microtubule-like structure formation, the number of nodes/high power field, and VEGF protein levels, demonstrating its anti-invasive capability.…”

“…In addition to the in vitro study, Wang et al [100] investigated the anticancer effects of neferine using an in vivo retinoblastoma model. WERI-Rb-1 cells were subcutaneously injected in female athymic nude mice, and the tumor-bearing animals were treated with 0.5-2 mg/kg of neferine every 3 days for 30 days.…”

Recent Developments in Search of Effective Herbal Medicine for The Treatment of Leukemia

“…Lotus plumule contains neferine, a bisbenzylisoquinoline alkaloid shown to have anti-cancer properties in past years [64,65]. Neferine suppressed the development of CML cell lines of K562 [66].…”

An updated review on pharmacological properties of neferine—A bisbenzylisoquinoline alkaloid from Nelumbo nucifera

“…Neferine (2 µM) effectively reduces angiogenesis, while inhibiting tumor development and invasion in WERI‐Rb‐1 retinoblastoma cells. Furthermore, neferine treatment inWERI‐Rb‐1 cells showed downregulation of nuclear protein Ki67 and vascular endothelial growth factor expression and facilitating apoptosis (Wang et al., 2020). Neferine (5 µM) prevents proliferative vitreoretinopathy in in vitro model of retinal pigment epithelial cells by inhibiting epidermal growth factor‐mediated cell division through cell cycle arrest at gap 1 phase (G1 phase) and epithelial mesenchymal transition (EMT) by increased expression of p21 and p27.…”

“…Neferine (10% & 20%) reduces the blood glucose and increased serum insulin levels with decreased lipid peroxidation and increased activities of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and GST in streptozotocin‐induced diabetic rats. The underlying molecular mechanism of wound healing activity of neferine was attributed through downregulation of inflammatory genes such as NF‐κβ, TNF‐α, IL‐6, and IL‐1β and activation of Nrf‐2 pathway (Wang et al., 2020). In myoblast L6 cells, neferine (50–150 μM) induced the expression of glucose transporter type 4 (GLUT4) thereby preventing insulin resistance.…”