بررسی اثر فرکشن های حلال چای کامبوچا بر تکثیر و القای آپوپتوز در سلول های سرطان کلورکتال رده ی HT-29

محورهای موضوعی : فصلنامه زیست شناسی جانوری

الهام قدوسی دهنوی ¹ (گروه زیست شناسی، دانشکده علوم، دانشگاه پیام نور، تهران، ایران)
محمد ارجمند ² (بخش بیوشیمی، انستیتو پاستور ایران، تهران، ایران)
رضا حاجی حسینی ³ (گروه زیست شناسی، دانشگاه پیام نور، مرکز تهران شرق، تهران، ایران)
زهرا زمانی ⁴ (بخش بیوشیمی، انستیتو پاستور ایران، تهران، ایران)
سیما نصری ⁵ (گروه زیست شناسی، دانشگاه پیام نور، مرکز تهران شرق، تهران، ایران)

تاریخ دریافت : 1399/03/05 تاریخ پذیرش : 1399/06/20 تاریخ انتشار : 1399/09/01

کلید واژه: آپوپتوز, چای کامبوچا, قطعه قطعه شدن DNA, کلوژنیک, ضد تکثیر,

چکیده مقاله :

نوشیدنی تخمیری که به طور سنتی حاصل تخمیر چای شیرین توسط قارچ کامبوچا است، دارای اثرات سودمندی در درمان بسیاری از بیماری ها به ویژه سرطان است. هدف از مطالعه ی حاضر بررسی اثرات فرکشن های مختلف چای بر میزان تکثیر و آپوپیتوز بر سلول های سرطان کلون، HT-29 است. در مطالعه تجربی حاضر، میزان بقاء و تکثیر سلولی فرکشن های حلال چای کامبوچا شامل فرکشن های کلروفرم، اتیل استات، بوتانول، هگزان و فاز آبی نهایی در غلظت های ( 0-900 میکروگرم بر میلی لیتر) بوسیله روش MTT و تست کلوژنیک مورد بررسی قرار گرفت. میزان آلقاء آپوپتوز توسط تست قطعه قطعه شدن DNA و روش فلوسایتومتری بررسی گردید. فرکشن های آبی و بوتانولی فاقد اثرات سمیت سلولی بودند. IC50 تعیین شده برای فرکشن های اتیل استات، کلروفرم و هگزان بعد از 24 ساعت به ترتیب 63/49 ± 1/213، 11/70 ± 2/296 و 29/83 ± 2/563 میکروگرم بر میلی لیتر تعیین شد. نتایج نشان داد که مهار رشد سلولی وابسته به دوز است. غلظت IC50 از این فرکشن ها سبب قطعه قطعه شدن DNA شد. با بررسی آپوپتوز به روش فلوسیتومتری با این غلظت ها نشان داد که جزء اتیل استات باعث بروز آپوپتوز سلولی شد، در حالی که مرگی که غلظت های IC50 از فرکشن های کلروفرم و هگزان ایجاد کرده بودند بیشتر از نوع نکروز بود. فرکشن اتیل استات چای کامبوچا به صورت وابسته به دوز باعث القاء مرگ سلولی از طریق مسیر آپوپتوز شد و ممکن است مسئول خاصیت ضد آپوپتوز و ضدتکثیر مشاهده شده باشند و به نظر می رسد می تواند کاندیدای مناسبی برای جلوگیری از تکثیر سلول های سرطانی کلورکتال باشد.

چکیده انگلیسی:

Fermented beverage, traditionally the result of the fermentation of sweet tea by Kombucha fungus, has beneficial effects in the treatment of many diseases, especially cancer. The present study is aimed at assessing the effect of different fractions of Kombucha tea on proliferation and apoptosis on colon cancer cell line HT-29. In the present experimental study, the survival rate and cell proliferation of Kumbucha tea solvent fractions including chloroform, ethyl acetate, butanol, hexane, and the final aqueous phase at concentrations of 0-900 μg/ ml were investigated by MTT and clonogenic assay. The rate of apoptosis induction was assessed by DNA fragmentation assay and flowcytometry. The aqueous and butanol fractions had no cytotoxic effects. After 24 hours, the IC50 for ethyl acetate, chloroform, and hexane was determined 213/49±1/63, 296/70±2/11 and 563/2±83/29 μg/ml, respectively. The results revealed that inhibition of cell growth was dose-dependent. The concentration of IC50 in these fractions caused the DNA to fragment. Apoptosis analysis by flowcytometry with these concentrations showed that the ethyl acetate component caused apoptosis while the death caused by IC50 concentrations of chloroform and hexane fractions was more than necrosis. The dose-dependent ethyl acetate fraction of Kambucha tea induced cell death through the apoptotic pathway and it was observed to be responsible for the anti-apoptotic and anti-proliferative properties. It seems to be a good candidate to prevent the proliferation of colorectal cancer cells.  

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