Scorpion venoms in gastric cancer.

Venom secretions from snakes, scorpions, spiders and bees, have been widely applied in traditional medicine and current biopharmaceutical research. Possession of anticancer potential is another novel discovery for animal venoms and toxins. An increasing number of studies have shown the anticancer effects of venoms and toxins of snakes, and scorpions in vitro and in vivo, which were achieved mainly through the inhibition of cancer growth, arrest of cell cycle, induction of apoptosis and suppression of cancer metastasis. However, more evidence is needed to support this concept and the mechanisms of anticancer actions are not clearly understood. The present review is focused on the recant updates on anticancer venom research.

Introduction

Venom is the secretion of venom glands found in certain snakes, scorpions, spiders, bees, lizards, cone snails and sea anemones (1). The venoms could be secreted in teeth, stingers, claws or even the skins of these animals, to paralyze and kill the prey or to protect themselves from predation and other dangers. The signs and symptoms after exposure to venoms vary from mild allergic reactions including itch, pain, swelling to respiratory arrest, paralysis, necrosis or even death (2). The utilization of animal venoms in folk medicine has been documented for a long time in some countries, such as China, India and the Middle East. For example, Chan Su, the dried toad venom from skin glands, first recorded in traditional Chinese medicine more than 1,000 years ago, has been long used as a diuretic, cardiotonic and anesthetic agent (3). Animal venoms are complex cocktails with various bioactive proteins/peptides and are variable between different species, making animal venoms a rich source for drug discovery. For the last 30 years, animal venoms and toxins have been widely investigated in the treatment of human disorders, such as diabetes, hypertension, chronic pain, HIV and cancer (4).

Scorpion venoms and toxins

Scorpion venoms are a complex mixture of water, salts, mucoproteins, lipids, nucleotides, glycoaminoglycans, histamine, serotonin, biogenic amines, low molecular weight (MW) peptides (e.g., neurotoxins), and high MW proteins (e.g., enzymes) (Fig. 1) (5). Each scorpion species has its own component profile in the venom and the number varies from dozens to hundreds. Small peptides (<10 kDa) are the most important components in scorpion venoms, which are believed to be responsible for intoxication and are widely investigated for biomedical and scientific applications. They are often considered as neurotoxins because the majority of peptides target and modify the ion channels of the excitable cells (e.g., neurons), which makes them valuable tools for ion channel research in neuroscience (5). Based on the types of targeted ion channels, scorpion venom peptides can be classified into four groups: Sodium (Na+), potassium (K+), calcium (Ca2+) and chloride (Cl−) channel toxins.

Figure 1.

Diagram of scorpion venom glands.

NaScTxs from scorpion venoms

Na+−channel scorpion toxins (NaScTxs) are polypeptides of 60–76 amino acid residues in length (6.5–8.5 kDa), tightly bound by four disulfide bridges. Current databases such as ArachnoServer (https://omictools.com/arachnoserver-tool) cover approximately 200 sequences of putative NaScTxs (6). The phyletic preference has been reported among NaScTxs, which principally classifies NaScTxs into two groups: ‘Classical’, highly active on mammalian Na+ channels; and ‘anti-insect’, highly active on insect Na+ channels. The latter toxins are subdivided into excitatory and depressant insect toxins (7).

KTxs from scorpion venoms

Scorpion venoms are rich sources of K+ channel toxins (KTxs), which block several types of K+ channels, e.g., voltage-gated K+ channels (Kv1.x), and Ca2+ activated K+ channels of small, intermediate and high conductance. KTxs are structurally categorized into four families: α-, β-, γ- and κ-KTxs, most of which share a conserved cysteine-stabilized α-helix and β-sheet structural motif (CSαβ) (8). The α-KTx family is considered as the largest in number, with approximately 140 peptides falling in 26 subfamilies, termed α-KT×1–26 and new peptides are described continuously (9). These peptides are composed of 23–42 amino acid residues with 3 or 4 disulfide bridges. These families of toxins are important blockers of Kv1.x and attract much attention in the studies of Kv channel structure-function and Kv-related channelopathies (10). The β-KTxs are long-chain peptides with 50–75 amino acid residues. The γ-KTxs are new interesting short chain peptides mainly targeting hERG channels, which are associated with the cell cycle and proliferation of several cancers (11).

Ca2+ and Cl− channel toxins from scorpion venoms

Different from NaScTxs and KTxs, scorpion venom peptides that target Ca2+ and Cl− channels, are scarcely known and have variable amino acid lengths (12). Imperatoxin A (IpTxa) proteins were purified from Pandinus imperator scorpion venom, and were also identified from other scorpion venoms, including maurocalcin, hemicalcin and hadrucalcin (7). Chlorotoxin (CTX/ClTx), a 36 amino acid small peptide purified from the Leiurus quinquestriatus scorpion venom, was initially described as a Cl− channel blocker that acts as a paralytic agent for small insects (13). A noteworthy finding and application of this toxin is that CTX can specifically bind to the Cl− channel on glioma cells and inhibit glioma progression (14). A phase II clinical trial is in progress with 131I-TM-601, a synthetic CTX coupled with radioactive iodine isotope (15).

Scorpion venom peptides with NDBPs

In addition to the ion channel-targeted peptides with disulfide bridges, there are a number of non-disulfide-bridged peptides (NDBPs) in scorpion venoms (16). These peptides show high diversity in structure and bioactivities. Currently, more than 40 scorpion venom NDBPs have been isolated and functionally described. The majority of NDBPs are antimicrobial peptides with a board spectrum of activity against bacteria, yeast, fungi and viruses (17). Hadruin, a 41 amino acid peptide isolated from Hadrurus aztecus, was the first to show antimicrobial activities at low micromolar concentrations (10–50 µM) (18). Mucroporin-M1, a modified antimicrobial peptide from scorpion Lychas mucronatus, was demonstrated to have antibacterial activity against antibiotic-resistant pathogens (19).

High MW enzymes

The high MW proteins in scorpion venoms are mainly diverse enzymes, which are believed to contribute to the venom cytotoxicity or potentiate the envenomation process. Therefore, a good understanding of these enzymes' structures and functions is useful for anti-venom strategy. In contrast to spider and snake venoms, scorpion venoms exhibit low levels of enzymatic activities (20). The enzymes present in scorpion venoms include hyaluronidase, phospholipase A2 (PLA2), L-amino acid oxidases (LAAOs) and proteases (21).

Hyaluronidase

Hyaluronidase can be found in several venomous species including snake, bee, spider and scorpion. This enzyme can degrade the hyaluronan, an extracellular matrix protein present in the soft connective tissues around blood vessels and increase the diffusion of toxins (22). Hyaluronidase has been purified from a few scorpion venoms, e.g., Heterometrus fulvipes, Tityus serrulatus and Palamneus gravimanus. BmHYA1, a hyaluronidase isolated from Mesobuthus martensi Karsch (BmK), was shown to remove hyaluronan and modulate the expression of CD44 variant in MDA-MB-231 breast cancer cells (23).

PLA2

PLA2 is a group of enzymes that hydrolyze the ester bonds of phospholipids into lysophospholipid, and fatty acids (24). The PLA2s described in scorpion venoms belong to sPLA2, which have a low MW (13–15 kDa) and are involved in tissue destruction and inflammation during the action of scorpionism (25). These enzymes have diverse biological and pharmacological potentials such as anti-coagulant and antibacterial activities (26).

Proteases

Proteases are important proteins in venoms that are involved in the post-translational processing of toxins and promote the spreading of toxins via degradation of matrix proteins. Two main types of proteases are identified in scorpion venoms: Serine proteases and metalloproteases (27). The first metalloprotease purified from scorpion venom was named antarease from the Brazilian scorpion Tityus serrulatus, which cleaves the vesicle-associated membrane proteins 2 and 8 (VAMP2 and VAMP8) (28). A serine protease-like protein (BMK-CBP) was also isolated from the Chinese red scorpion BmK (29).

LAAOs from scorpion and snake venoms

LAAOs are a group of flavoenzymes that catalyze oxidative deamination of L-amino acid substrates and form the corresponding α-keto acids, hydrogen peroxide and ammonia (30). LAAOs could be widely found in nature, including bacteria, fungi, seaweeds and snake venoms.

The presence of LAAOs in scorpion venoms is not widely reported but its activity is observed in the Chinese red scorpion venom BmK (31). Snake venoms are the richest source of LAAOs, which are responsible for the yellowish colour for the venoms. Recently, LAAOs have become a research interest in biomedicine because they have multi-effects including anti-microbial, anti-HIV, anti-coagulant, apoptosis-inducing, edema-inducing and hemorrhagic activities (32). Notably, some snake venom LAAOs can induce platelet aggregation, such as LAAOs from B. moojeni, Bothrops atrox and Trimeresurus jerdonii. While LAAOs from snake venoms of Vipera berus, Naja oxiana and others, were reported to inhibit platelet aggregation. The application of LAAOs in cancer research is another recent scientific attempt, by applying the cytotoxic effects of H2O2 generated from LAAO enzymatic reaction.

The anticancer potential of scorpion venoms and toxins

Scorpion envenomation is a risk for public health in tropical and subtropical regions and there is a clear need for the improvement in specific (anti-venom) and systematic treatments. Scorpions and scorpion venoms have been applied in traditional medicine for long periods in China, India and Africa (33). Additionally, scorpion venoms have antimicrobial functions against bacteria, fungi, yeasts and viruses. Studies showed that scorpion venom-derived protein mucroporin-M1 inhibited the amplification of hepatitis virus B and another peptide Kn2–7 possesses anti-HIV-1 activity (34).

The anticancer potential is another recently observed biological property of scorpion venoms and toxins. A number of experimental and preclinical studies have shown that scorpion venoms and toxins could impair cancer growth, induce apoptosis and inhibit cancer metastasis in vitro and in vivo. Several active molecules with confirmed anticancerous activities such as proliferation inhibition, cell cycle arrest, induction of apoptosis and decreasing cell migration have been purified from scorpion venoms. The investigated cancer types included glioma, neuroblastoma, leukemia, lymphoma, breast, lung and prostate cancers (35).

Among all the scorpions tested in cancer research, the BmK scorpion venom is probably the first to be reported to possess antitumor properties. BmK scorpion venom is the most extensively studied in China and several active molecules have been isolated and characterized.

Polypeptide extract from the scorpion venom (PESV), a group of partially purified polypeptides with 50–60 amino acids from the crude venom of BmK, was reported to inhibit cell proliferation and induce cell apoptosis of DU 145 human prostate cancer cells (36). An analgesic-antitumor peptide (AGAP) isolated from a fusion protein SUMO-AGAP, which connected a small ubiquitin-related modifier to AGAP, inhibited cell proliferation and migration of SHG-44 human malignant glioma cells via interfering with the p-AKT, NF-κB, BCL-2, and MAPK signaling pathways (37). The most notable evidence regarding the anticancer effects of scorpion venoms comes from CTX utilized in the treatment of glioma. Based on electrophysiological evidence, the Cl− channel was initially considered to be responsible for the affinity and specificity of CTX to glioma. However, further studies by protein interaction approaches with a recombinant His-CTX, revealed that the principle receptor of CTX is matrix metalloproteinase-2 (MMP-2), a protease that is over-expressed on the surface of glioma cells (38).

Conclusions

It can be concluded that scorpion venoms hold great potential in their actions against cancer cells via targeting the ion channels to inhibit cell proliferation and metastasis, secondly via induction of apoptosis by cell cycle arrest, caspase activation, and mitochondria depolarization or oxidative stress. Nevertheless, as a novel research field, more efforts should be made to extensively evaluate the anticancer effects of scorpion venoms and toxins to understand the mechanisms of action.