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Iranian Journal of Biotechnology، جلد ۱۴، شماره ۳، صفحات ۱۲۵-۱۳۱
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عنوان انگلیسی Synergistic Effect of Expressed miR-128 and Puma protein on Targeted Induction of Tumor Cell Apoptosis
چکیده انگلیسی مقاله Background: Puma is a highly robust pro-apoptotic protein. The protein becomes activated by p53 ensuing beyond-repair DNA damage. Downregulation of SIRT 1, by miR-128, elevates activated p53 that foment Puma indirectly. Objectives: In the present study, we used two-expression Adeno-Associated Virus (AAV) system for co-expression of miR-128 and Puma in order to evaluate apoptotic response; both in the tumor and normal cells, respectively. Materials and Methods: Three recombinant AAVs constructs were generated. The First rAAV bearing Puma under the control of hTERT (p-AAV), the second construct designed such that to carry miR-128 downstream of CMV (mi-AAV), and the last construct comprises of the both CMV-miR-128 and hTERT- Puma. Real-Time PCR and western blotting were used to evaluate expression levels of the transduced genes. Results: MTT assay and DAPI staining shown suicidal effect of each recombinant AAV vectors. p-AAV cytotoxicity was recorded for 62% of the tumor cells, while for normal cells it was only 20% cytotoxic. The second construct, mi-AAV, was not as potent and selective as p-AAV. This construct was shown to be 27% and 16% cytotoxic for BT-474 and HEK-293 cells, respectively. Co-expression of Puma and miR-128 (p-mi-AAV) was accomplished with a selective cytotoxicity toward BT-474. This construct was 85% toxic for tumor cells, although it was only 25% toxic for the normal cell line (HEK-293). Conclusions: In this study, we have shown that not only Puma is able to instigate apoptotic response but also its co-expression along with miR-128 could significantly enhance apoptosis in a synergistic manner.
کلیدواژه‌های انگلیسی مقاله AAV,Adeno-Associated Virus,Gene therapy,Puma,Suicide gene
نویسندگان مقاله شهریار خوش طینت nikkhoi | khoshtinat nikkhoi
department of medical biotechnology, faculty of medical sciences, tarbiat modares university, tehran, iran
سازمان اصلی تایید شده: دانشگاه تربیت مدرس (Tarbiat modares university)

روح الله درستکار |
applied virology research center, baqiyatallah university of medical sciences, tehran, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی بقیه الله (Baqiyatallah university of medical sciences)

سعید رنجبر |
department of medical biotechnology, faculty of medical sciences, tarbiat modares university, tehran, iran
سازمان اصلی تایید شده: دانشگاه تربیت مدرس (Tarbiat modares university)

هدیه حیدرزاده |
department of microbiology, azad university of shahreh-qods, tehran, iran

مهدی تات |
applied virology research center, baqiyatallah university of medical sciences, tehran, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی بقیه الله (Baqiyatallah university of medical sciences)

مجدالدین قلاوند |
applied virology research center, baqiyatallah university of medical sciences, tehran, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی بقیه الله (Baqiyatallah university of medical sciences)

علیرضا فراست |
department of medical biotechnology, faculty of medical sciences, tarbiat modares university, tehran, iran
سازمان اصلی تایید شده: دانشگاه تربیت مدرس (Tarbiat modares university)

محمد صادق هاشم زاده | mohammad sadegh
applied virology research center, baqiyatallah university of medical sciences, tehran, iran
سازمان اصلی تایید شده: دانشگاه علوم پزشکی بقیه الله (Baqiyatallah university of medical sciences)

نشانی اینترنتی http://www.ijbiotech.com/article_15461_a48f3a8540f80a631f5b25293bad0b9d.pdf
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