Heterologous expression of biologically active Mambalgin-1 peptide as a new potential anticancer, using a PVX-based vira
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ORIGINAL ARTICLE
Heterologous expression of biologically active Mambalgin‑1 peptide as a new potential anticancer, using a PVX‑based viral vector in Nicotiana benthamiana Ghaffar Khezri1 · Bahram Baghban Kohneh Rouz1 · Hamideh Ofoghi2 · Seyed Javad Davarpanah3 Received: 19 August 2019 / Accepted: 30 April 2020 © Springer Nature B.V. 2020
Abstract Mambalgin-1 is a peptide that acts as a potent analgesic through inhibiting acid-sensing ion channels (ASIC) in nerve cells. Research has shown that ASIC channels are involved in the proliferation and growth of cancer cells; therefore, Mambalgin-1 can be a potential anti-cancer by inhibiting these channels. In the present study, the Nicotiana benthamiana codon optimized Mambalgin-1 gene was synthesized and cloned in PVX (potato virus X) viral vector. The two cultures of Agrobacterium containing Mambalgin-1 and P19 silencing suppressor genes were co-agroinfiltrated into N. benthamiana leaves. Five days post infiltration, the production of recombinant Mambalgin-1 was determined by western blotting. For biological activity, MTT (3(4, 5-dimethylthiazole-2-yl)-2, 5-diphenyltetrazolium bromide) was analyzed for the cytotoxicity recombinant Mambalgin-1 from the transformed plants on nervous (SH-SY5Y) and breast (MCF7) cancer cells. The results showed that the plants expressing open reading frame of Mambalgin-1 showed recombinant 7.4 kDa proteins in the entire plant extract. In the MTT test, it was found that Mambalgin-1 had cytotoxic effects on SH-SY5Y cancer cells, yet no effects on MCF7 cancer cells were observed. According to the results, the expression of the biologically active recombinant Mambalgin-1 in the transformed plant leaves was confirmed and Mambalgin-1 can also have anti-cancer (inhibition of ASIC channels) potential along with its already known analgesic effect. Key message The Mambalgin-1 transiently expressed in Nicotiana benthamiana plant and its anti-cancer effect on SH-SY5Y nerve cancer cells, since ASIC channels are involved in the proliferation of cancer cells, was confirmed.
Communicated by Degao Liu. Electronic supplementary material The online version of this article (doi:https://doi.org/10.1007/s11240-020-01838-x) contains supplementary material, which is available to authorized users. * Bahram Baghban Kohneh Rouz [email protected] * Seyed Javad Davarpanah [email protected] 1
Department of Plant Breeding and Biotechnology, University of Tabriz, Tabriz, Iran
2
Department of Biotechnology, Iranian Research Organization for Science and Technology, Tehran, Iran
3
Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran
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Vol.:(0123456789)
Plant Cell, Tissue and Organ Culture (PCTOC)
Keywords Mambalgin-1 · Transient expression · Nicotiana benthamiana · Agroinfiltration · Acid sensing ion channels (ASICs) · Cancer
Introduction In 2012, Diochot et al. discovered a pair of peptides, mambalgin-1 and mambalgin-2, from the venom of black mamba snakes (Dendroaspis polylepsis) which had
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