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Gene Therapy To Kill Tumor Cells Effectively

928 words - 4 pages

Recent studies are focusing on targeted gene therapies to kill tumor cells effectively without adverse effects on other normal cells (27). Melanoma differentiation-associated gene-7 (mda-7) gene is a novel tumor suppressor (2), that induces apoptosis in transformed cells without negatively affecting normal cells (28). Several studies have been performed to investigate its potential therapeutic effects and also to improve its functional properties (13).
Here, we constructed three plasmids, including standard or fusion forms of mda-7 beside RGD peptides then their expression examined in some human cell lines. To investigate the effect of aforementioned fusion peptides, pIRES/mda-7, ...view middle of the document...

Xiao et al fused RGD peptide beside mda-7 protein for the first time. Their group added RGD-4C to the N-terminus of IL-24 then demonstrated that both simple and fusion protein have similar behavior in apoptosis induction and adhesion experiments for MCF-7 cancer cell (19). Here, in order to enhance mda-7/IL-24 cell adhesion rather than elevating its inherent apoptotic inducing activity by improving its bystander effect, as a first effort two kinds of RGD motifs introduced at the C-terminal of mda-7/IL-24 then their expressions were evaluated in both normal and tumor cell line. This kind of design would be helpful for gene based therapy and will get rid of protein processing in bacterial expression. Of note, this strategy will exploit both intracellular and extracellular route of apoptosis through endogenous expression and subsequent secretion into surrounding cells respectively.
Of 25 known integrins, eight type bind to RGD (Arg-Gly-Asp) peptide which serves as the main integrin recognition site in the extracellular matrix proteins (29). Hence fusing RGD peptides supposed to augment tumor cell targeting and show higher apoptotic rate in transfected cells through positive feedback loop. Integrin α5β3 has been shown to be expressed on a variety of hepatocellular lines like Hep-G2 but present only in negligible level in Huh-7 line (30). So here to explore intracellular expression and avoid extracellular effect of mda-7 low α5β3 integrin cell, Huh-7 used in our project.
Our selected RGD motifs had fine differences in sequence as original cyclic RGD-4C (ACDCRGDCFCG) harbour at least 2 more terminal amino-acid compared to smaller version. Theoretically, original size of mda-7 may exhibits immune induction which will be avoided in our smaller RGD2 version (CDCDGRCFC). This kind of design will avoid immune induction albeit keeping the targeting potency of...

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