CONSTRUCTION OF EXPRESSING VECTORS INCLUDING MELANOMA DIFFERENTIATION‐ASSOCIATED GENE‐7 (MDA‐7) FUSED WITH THE RGD SEQUENCES FOR BETTER TUMOR TARGETING

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KHODADAD MAHBOOBEH; HOSSEINI SEYED YOUNES; SHENAVAR FATEMEH; ERFANI NASROLLAH; BINA SAMANEH; AHMADIAN SHAHIN; FATTAHI MOHAMMAD REZA; HAJHOSSEINI REZA

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Journal: Iranian Journal of Basic Medical Sciences Year:2015 | Volume:18 | Issue:8 Page(s): 780-787

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Title

CONSTRUCTION OF EXPRESSING VECTORS INCLUDING MELANOMA DIFFERENTIATION‐ASSOCIATED GENE‐7 (MDA‐7) FUSED WITH THE RGD SEQUENCES FOR BETTER TUMOR TARGETING

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Keywords

MDA‐7

RGD PEPTIDE

TUMOR TARGETING

Abstract

Objective (s): Up to now, many researches have been performed to improve the antitumoral effect of melanoma differentiation‐associated gene‐7 (MDA‐7) protein. The purpose of our research was to construct 3 expression vectors producing MDA‐7 in fusion with RGD (Arginine‐Glycine‐Aspartic acid) peptide and evaluate their expression.Materials and Methods: MDA‐7 gene with two different RGD sequences was amplified by PCR then was cloned by TA-loning system. The colonies including these genes were selected by blue–hite screening, colony PCR, and sequencing, respectively. Afterward, the genes were sub‐cloned into the expression vector following confirmation by colony PCR and sequencing. In addition, these constructs were transfected into 293 and Huh‐7 cells for further expression analysis. The MDA‐7 gene expression was evaluated by RT‐PCR and IF (immunofluorescence assay). DNA laddering test and trypan blue exclusion assays were performed to screen cytotoxicity of prepared plasmids.Results: Three different MDA‐7 genes with terminal RGD PEPTIDE were cloned correctly into the expression vectors and their expression was confirmed to be suitable by RT‐PCR and IF assay. It was shown that expressions were limited to those transfected, GFP shining cells. No significant cytotoxicity was observed by simple assays in all plasmid treated cells. In expressing cells, all forms of MDA‐7 protein were localized mainly around ER prenuclear compartment while GFP protein was distributed evenly among them.Conclusion: Theoretically RGD tagged MDA‐7 would be able to induce apoptosis with more specificity and stronger than the standard one, therefore, these new constructs may have the potential for further researches.

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APA: Copy

KHODADAD, MAHBOOBEH, HOSSEINI, SEYED YOUNES, SHENAVAR, FATEMEH, ERFANI, NASROLLAH, BINA, SAMANEH, AHMADIAN, SHAHIN, FATTAHI, MOHAMMAD REZA, & HAJHOSSEINI, REZA. (2015). CONSTRUCTION OF EXPRESSING VECTORS INCLUDING MELANOMA DIFFERENTIATION?ASSOCIATED GENE?7 (MDA?7) FUSED WITH THE RGD SEQUENCES FOR BETTER TUMOR TARGETING. IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES, 18(8), 780-787. SID. https://sid.ir/paper/669833/en

Vancouver: Copy

KHODADAD MAHBOOBEH, HOSSEINI SEYED YOUNES, SHENAVAR FATEMEH, ERFANI NASROLLAH, BINA SAMANEH, AHMADIAN SHAHIN, FATTAHI MOHAMMAD REZA, HAJHOSSEINI REZA. CONSTRUCTION OF EXPRESSING VECTORS INCLUDING MELANOMA DIFFERENTIATION?ASSOCIATED GENE?7 (MDA?7) FUSED WITH THE RGD SEQUENCES FOR BETTER TUMOR TARGETING. IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES[Internet]. 2015;18(8):780-787. Available from: https://sid.ir/paper/669833/en

IEEE: Copy

MAHBOOBEH KHODADAD, SEYED YOUNES HOSSEINI, FATEMEH SHENAVAR, NASROLLAH ERFANI, SAMANEH BINA, SHAHIN AHMADIAN, MOHAMMAD REZA FATTAHI, and REZA HAJHOSSEINI, “CONSTRUCTION OF EXPRESSING VECTORS INCLUDING MELANOMA DIFFERENTIATION?ASSOCIATED GENE?7 (MDA?7) FUSED WITH THE RGD SEQUENCES FOR BETTER TUMOR TARGETING,” IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES, vol. 18, no. 8, pp. 780–787, 2015, [Online]. Available: https://sid.ir/paper/669833/en

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