title: Construction and characterization of gelonin and saporin plasmids for toxic gene-based cancer therapy.
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.
authors.affiliation: College of Pharmacy and Research Institute of Pharmaceutical Sciences, Gyeongsang National University, 501 Jinju Daero, Jinju, Gyeongnam, 52828, Republic of Korea.|College of Pharmacy, Inje University, 197 Injero, Gimhae, Gyeongnam, 50834, Republic of Korea.
authors.forename: Kyoung Ah
authors.initials: KA
authors.lastname: Min
authors.affiliation: Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnosis, School of Pharmacy, Tianjin Medical University, Tianjin, 300070, China.
authors.forename: Huining
authors.initials: H
authors.lastname: He
authors.affiliation: Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnosis, School of Pharmacy, Tianjin Medical University, Tianjin, 300070, China.|Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, 428 Church St, Ann Arbor, MI, 48109, USA.
authors.forename: Victor C
authors.initials: VC
authors.lastname: Yang
authors.affiliation: College of Pharmacy and Research Institute of Pharmaceutical Sciences, Gyeongsang National University, 501 Jinju Daero, Jinju, Gyeongnam, 52828, Republic of Korea. shinmc@gnu.ac.kr.
authors.forename: Meong Cheol
authors.initials: MC
authors.lastname: Shin
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids / Q000235:genetics*; D011094:Polyethyleneimine / Q000737:chemistry; D051381:Rats; D054789:Ribosome Inactivating Proteins, Type 1 / Q000235:genetics*; D000076985:Saporins; D014162:Transfection
chemical_list: D054789:Ribosome Inactivating Proteins, Type 1; D049452:Green Fluorescent Proteins; C025674:GEL protein, Gelonium multiflorum; D011094:Polyethyleneimine; D000076985:Saporins
keywords: Gelonin; Gene therapy; Ribosome inactivating protein; Saporin; Tumor