Neferine, an alkaloid from lotus seed embryo targets HeLa and SiHa cervical cancer cells via pro‐oxidant anticancer mechanism

Abstract

Apoptosis and autophagy are important processes that control cellular homeostasis and have been highlighted as promising targets for novel anticancer drugs. This study aims to investigate the inhibitory effects and mechanisms of Neferine (Nef), an alkaloid from the lotus seed embryos of Nelumbo nucifera (N. nucifera), as a dual inducer of apoptosis and autophagy through the reactive oxygen species (ROS) activation in cervical cancer cells. Nef and N. nucifera extract suppressed the cell viability of HeLa and SiHa cells in a dose‐dependent manner. Importantly, Nef showed minimal toxicity to normal cells. Furthermore, Nef inhibited anchorage‐independent growth, colony formation and migration ability of cervical cancer cells. Nef induces mitochondrial apoptosis by increasing pro‐apoptotic protein bax, cytochrome‐c, cleaved caspase‐3 and caspase‐9, poly‐ADP ribose polymerase (PARP) cleavage, DNA damage (pH2AX) while downregulating Bcl‐2, procaspase‐3 and procaspase‐9, and TCTP. Of note, apoptotic effect by Nef was significantly attenuated in the presence of N‐acetylcysteine (NAC), suggesting pro‐oxidant activity of this compound. Nef also promoted autophagy induction through increasing beclin‐1, atg‐4, atg‐5 and atg‐12, LC‐3 activation, and P62/SQSTM1 as determined by western blot analysis. Collectively, these results demonstrate that Nef is a potent anticancer compound against cervical cancer cells through inducing apoptosis and autophagic pathway involving ROS.

Lotus ( Nelumbo nucifera ) seedpod extract inhibits cell proliferation and induces apoptosis in non‐small cell lung cancer cells via downregulation of Axl

“…As the molecular mechanisms associated with the anticancer ac- Bcl-2, respectively (Dasari et al, 2020;Zhao et al, 2017). In NSCLC cells, neferine was evaluated as an apoptosis inducer through reactive oxygen species (ROS) generation (Kalai Selvi et al, 2017;Poornima et al, 2013) and MAPKs' activation (An et al, 2020).…”

“…For example, the leaf extract of lotus inhibited proliferation and metastasis of breast cancer cells (Yang, Hung, et al., 2019) and the stamen extract was found to be cytotoxic to colon cancer cells (Zhao et al., 2017). Neferine, an alkaloid derived from green seed embryos of lotus has been shown to have antiproliferative potential in hepatocellular carcinoma (Yoon et al., 2013), ovarian cancer (Xu et al., 2016), renal cancer (Kim et al., 2019), cervical cancer (Dasari et al., 2020), esophageal squamous cell carcinoma (An et al., 2020), osteosarcoma cells and NSCLC cells (Poornima et al., 2013). Flavonoids from leaf extract of lotus was also reported to inhibit growth of breast cancer cells as well as xenografts in mouse model (Chang et al., 2016).…”

“…As the molecular mechanisms associated with the anticancer activities of extracts or constituents prepared from lotus, the induction of apoptosis via targeting different intracellular molecules involved in various signaling pathways has been demonstrated in many cancers. For example, stamen extract of lotus induced apoptosis of colon carcinoma cell by increase or decrease of expression of apoptosis‐associated genes such as Fas and Fas ligand or anti‐apoptotic gene such as Bcl‐2, respectively (Dasari et al., 2020; Zhao et al., 2017). In NSCLC cells, neferine was evaluated as an apoptosis inducer through reactive oxygen species (ROS) generation (Kalai Selvi et al., 2017; Poornima et al., 2013) and MAPKs’ activation (An et al., 2020).…”

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

“…Neferine (5–50 μM) treatment in HeLa and SiHa cervical cancer cell lines induces mitochondrial‐mediated apoptosis through overexpression of bax, cytochrome‐c, cleaved caspase‐3 and caspase‐9, cleaved PARP, and phosphorylated‐H2A histone family member X (pH2AX). Increased autophagic flux was observed after neferine treatment by the increased expression levels of beclin‐1, autophagy‐related gene (atg) ‐5, ‐4, and ‐12, LC‐3 II (microtubule‐associated protein 1A/1B‐light chain‐3 II), and P62 (Dasari et al., 2020). Strong anticancer and anti‐metastatic efficacy were reported with neferine (4 μM) against GIST‐T1 (human gastrointestinal stromal tumor) and SGC7901 cell line (human gastric cancer) by activating microRNA‐449 (miR‐449a) and downregulating p‐PI3K, phsophso‐serine/threonine kinase 1 (p‐AKT), and Notch 1, 2 & 3 protein expressions (Xue et al., 2019).…”

Targeting Autophagy with Natural Products as a Potential Therapeutic Approach for Cancer

“…Neferine induces ROS-dependent mitochondrial mediated apoptosis by increasing the expression of proapoptotic proteins Bax, cytochrome c, cleaved caspase-3 and caspase-9. It also activates autophagy via increasing Beclin1, ATG4, ATG5 and ATG12, LC3-II expression levels in cervical cancer cells [ 269 ]. Additionally, neferine enhances cisplatin-induced autophagic cell death in human lung adenocarcinoma via downregulation of the PI3K/AKT/mTOR signaling pathway [ 270 ].…”