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Construction and characterization of gelonin and saporin plasmids for toxic gene-based cancer therapy

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Abstract

Toxic gene therapy (or suicidal gene therapy) is gaining enormous interest, specifically for the treatment of cancer. The success of this therapy lies in several crucial factors, including the potency of gene products to kill the transfected tumor cells and the transfection ability of the transfection vehicles. To address the potency problem, in the present study, we engineered two separate mammalian transfection plasmids (pSAP and pGEL) containing genes encoding ribosome inactivating proteins (RIPs), gelonin and saporin. After the successful preparation and amplification of the plasmids, they were tested on various cancer cell lines (HeLa, U87, 9L, and MDA-MB-435) and a noncancerous cell line (293 HEK) using polyethyleneimine (PEI) as the transfection agent. Transfection studies performed under varying gene concentration, incubation time, and gene-to-PEI ratios revealed that, compared to the treatment of pGFP (GFP expression plasmid)/PEI, both pGEL/PEI and pSAP/PEI complexes could induce significantly augmented cytotoxic effects at only 2 μg/mL gene concentration. Importantly, these cytotoxic effects were observed universally in all tested cancer cell lines. Overall, this study demonstrated the potential of pGEL and pSAP as effective gene candidates for the toxic gene-based cancer therapy.

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Acknowledgments

This research was supported by grants from Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF-2015R1A6A3A01020598 & NRF-2015R1C1A1A02036781). In addition, this work was partially supported by the NSFC 2013 A3 Foresight Program (81361140344), National Key Basic Research Program of China (2013CB932502), and by National Institutes of Health R01 Grants CA114612. This work was also supported in part by the National Natural Science Foundation of China (NSFC, 81402856).

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Authors and Affiliations

  1. College of Pharmacy and Research Institute of Pharmaceutical Sciences, Gyeongsang National University, 501 Jinju Daero, Jinju, Gyeongnam, 52828, Republic of Korea

    Kyoung Ah Min & Meong Cheol Shin

  2. College of Pharmacy, Inje University, 197 Injero, Gimhae, Gyeongnam, 50834, Republic of Korea

    Kyoung Ah Min

  3. Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnosis, School of Pharmacy, Tianjin Medical University, Tianjin, 300070, China

    Huining He & Victor C. Yang

  4. Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, 428 Church St, Ann Arbor, MI, 48109, USA

    Victor C. Yang

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  1. Kyoung Ah Min
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Correspondence to Meong Cheol Shin.

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Min, K.A., He, H., Yang, V.C. et al. Construction and characterization of gelonin and saporin plasmids for toxic gene-based cancer therapy. Arch. Pharm. Res. 39, 677–686 (2016). https://doi.org/10.1007/s12272-016-0739-3

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