بررسی اثر Avastin بر آپوپتوز سلول های همکشتی داده شده HepG2 بر هیدروژل حاوی سلول HUVEC از طریق عملکرد مسیر مولکولی P53 و تغییرات سطح گونه های اکسیژنی اکسایشگر
محورهای موضوعی :
الهام نقی موسوی قزلجه
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هانیه جعفری
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زهرا کیان مهر
3
1 - گروه زیست شناسی، دانشکده علوم و فناوری های همگرا، دانشگاه آزاد اسلامی، واحد علوم و تحقیقات، تهران، ایران
2 - گروه زیست شناسی، واحد علوم و تحقیقات تهران، دانشگاه آزاد اسلامی، تهران، ایران
3 - ﮔﺮوه بیوشیمی، دانشکده علوم زیستی، دانشگاه آزاد اسلامي، واحد تهران شمال، تهران، ايران
کلید واژه: آنتی اکسیدان, داربست هیدروژلی, سرطان, کبد, آپوپتوز,
چکیده مقاله :
کارسینوم هپاتوسلولار پنجمین سرطان شایع در سراسر جهان می باشد که با عوارض شدید همراه است. در این پژوهش، بررسی اثر داروی آواستین بر تکثیر سلولهای HepG2 از طریق فعال شدن مسیر P53 و تغییر محتوای آنتی‌اکسیدانی سلولهای HepG2 کشت داده شده بر داربست هیدروژلی حاوی سلولهای HUVEC انجام شد. سلولهای HepG2 بر روی داربست هیدروژلی کلاژن نوع I حاوی HUVEC کشت داده شدند و پس از تیمار با غلظت‌های مختلف آواستین، زنده مانی سلول‌ها، با استفاده از تست MTT و میزان آپوپتوز با استفاده از تکنیک فلوسایتومتری انجام شد. میزان پراکسیداسیون لیپیدی، فعالیت آنزیمی سوپراکسید دیسموتاز (SOD)، وضعیت اکسیدانی کل (TOS) و ظرفیت آنتی‌اکسیدانی کل (TAS) با استفاده از روش کالرومتریک و میزان پروتئین p53 با استفاده از تکنیک ایمونوسیتوشیمی انجام شد. بررسی نتایج نشان می دهد که میزان آپوپتوز در سلولهای HepG2 همکشتی شده با HUVEC-Scaffold و داروی آواستین نسبت به سایر گروه‌ها بیشتر است. نتایج، نشان دهنده افزایش معنی دار میزان اکسیدان کل و مالون دی آلدئید و کاهش معنی دار میزان فعالیت آنزیم سوپراکسید دیسموتاز و ظرفیت آنتی اکسیدان کل در دو گروه HepG2+Scaffold-HUVEC و HepG2+Scaffold تیمار شده با داروی آواستین می باشد. داروی آواستین باعث افزایش درصد میزان p53 در سلولهای HepG2 تحت تیمار همکشتی شده با Scaffold و Scaffold-HUVEC شود. داربست هیدروژلی به عنوان یک گزینه برای شبیه سازی بهینه زندگی سلولهای سرطانی استفاده و نشان داده شد که آواستین توانایی تأثیر ضد سرطانی شایانی در این ساختارها دارد.
Hepatocellular carcinoma is the fifth most common cancer worldwide, which is associated with severe complications. In this study, the effect of Avastin drug on the proliferation of HepG2 cells was investigated through the activation of the P53 pathway and the change of the antioxidant content of HepG2 cells cultured on a hydrogel scaffold containing HUVEC cells. In this study, HepG2 cells were cultured on type I collagen hydrogel scaffolds containing HUVEC and after treatment with different concentrations of Avastin, cell viability was measured using the MTT test and the rate of apoptosis using flow cytometry. The level of lipid peroxidation, superoxide dismutase (SOD) enzyme activity, total oxidant status (TOS) and total antioxidant capacity (TAS) were measured using calorimetric method and p53 protein level was measured using immunocytochemistry technique. Examining the results shows that the rate of apoptosis in HepG2 cells co-cultured with HUVEC-Scaffold and Avastin drug is significantly higher than other groups. The results show a significant increase in the amount of total oxidant and malondialdehyde and a significant decrease in the activity of the superoxide dismutase enzyme and total antioxidant capacity in the two groups of HepG2+Scaffold-HUVEC and HepG2+Scaffold treated with Avastin. Avastin can increase the percentage of p53 in HepG2 cells co-cultured with Scaffold and Scaffold-HUVEC. In this study, hydrogel scaffold was used as an option for optimal simulation of the life of cancer cells and it was shown that the drug Avastin has the ability to have a good anti-cancer effect in these structures.
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