Anemone blanda’nın sağlıklı (Beas-2B) ve kanser (A549) akciğer epitel hücre hatları üzerindeki in vitro antiproliferatif ve antioksidan etkilerinin araştırılması
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Date
2022
Authors
İrkören, Sinem Fındık
Journal Title
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Volume Title
Publisher
Bursa Uludağ Üniversitesi
Abstract
Kanser, tüm dünyada morbidite ve mortalitenin önde gelen nedenlerinden biri olmaya devam etmektedir. Kanser, bulaşıcı olmayan hastalıklar dışında, kardiyovasküler hastalıklardan sonra ikinci önde gelen ölüm nedenidir. Son yıllarda, çeşitli bitkisel ürünlerin farklı kanserlere karşı umut verici anti-kanser etkinliği gösterilmiştir. Bu bağlamda tez çalışmasında, Anemone blanda bitkisinden elde edilen metanolik ekstraktlarının anti-kanser potansiyellerinin araştırılması amaçlanmıştır. Bu amaca yönelik olarak, insan sağlıklı bronşiyal epitel hücre hattı olan Beas-2B ve insan akciğer karsinoma hücre hattı olan A549 hücreleri kullanılmıştır. Sitotoksisitenin belirlenebilmesi için XTT testi ve Klononjenik testi, DNA hasarlarının belirlenebilmesi için Komet yöntemi ve hücre içi reaktif oksijen seviyesine etkilerini belirlemek için ROS testi kullanılmıştır. XTT testi sonucunda Beas-2B ve A549 hücre hatlarındaki IC50 değeri sırasıyla 21,90 μg/mL ve 21,25 μg/mL olarak, klonojenik testte ise bu değerler Beas-2B ve A549 hücre hatları için sırasıyla 24,57 μg/mL ve 27,62 μg/mL olarak belirlenmiştir. Komet testinde ise XTT ve Klonojenik testinde kullanılarak dozlara ek olarak H202’li kombine dozlarda eklenmiştir. Beas-2B ve A549 hücre hatlarındaki kuyruk uzunluğu, kuyruk % DNA ve olive kuyruk momenti hesaplanmış ve elde edilen sonuçlarda DNA iplik kırıklarında kontrol grubuna göre istatistiksel olarak anlamlı artışlar olduğu belirlenirken H202’li dozlarda istatistiksel olarak azaldığı belirlenmiştir. ROS testinde komet testindeki aynı dozlar kullanılırken sonuçlarında ise Beas-2B için hücre içi ROS seviyesinde istatistikî olarak anlamlı bir değişiklik görülmemiştir. Kombine dozlarda ise Anemone ekstraktının ortalama hücre içi ROS seviyesini 40 μg/mL dozdan itibaren istatistikî olarak anlamlı şekilde azalttığı görülmüştür. A549 için ROS seviyesini istatistikî olarak anlamlı şekilde düşürürken H2O2 kombine dozlarda hücre içi ROS seviyesini istatistiksel anlamlı olarak düşürdü fakat 100μg/ml+1000 μM H2O2’lik kombine dozda ekstrakt H2O2'in etkisini arttırarak hücre içi ROS seviyesini istatistikî olarak anlamlı şekilde arttırdığı görüldü.
Cancer continues to be one of the leading causes of morbidity and mortality worldwide. Cancer is the second leading cause of death after cardiovascular diseases, excluding non-communicable diseases. Today, research on various herbal products is increasingly continuing to develop new drugs against diseases such as cancer. In recent years, promising anti-cancer efficacy of various herbal products against different cancers has been demonstrated. In this context, in the thesis study, it was aimed to investigate the anti-cancer potentials of methanolic extracts obtained from Anemone blanda plant. For this purpose, the human healthy bronchial epithelial cell line Beas-2B and the human carcinoma cell line A549 cells were used. XTT test and Clonogenic test were used to determine cytotoxicity, Komet method to determine DNA damages and ROS test to determine their effects on intracellular reactive oxygen level. As a result of the XTT test, the IC50 values in Beas-2B and A549 cell lines were determined as 21.90 μg/mL and 21.25 μg/mL, respectively. In the clonogenic test, these values were determined as 24.57 μg/mL and 27.62 μg/mL for Beas-2B and A549 cell lines, respectively. In the Comet test, it was used in XTT and Clonogenic tests, in addition to the doses, it was added in combined doses with H2O2. Tail length, tail % DNA and olive tail moment in Beas-2B and A549 cell lines were calculated, and it was determined that there were statistically significant increases in DNA strand breaks compared to the control group, while it was statistically decreased at doses with H2O2. While the same doses as in the comet test were used in the ROS test, there was no statistically significant change in the intra-ROS level for Beas-2B in the results. In combined doses, it was observed that the mean intracellular ROS level of Anemone extract was statistically significantly reduced from 40 μg/mL dose, while it statistically decreased the ROS level for A549, while it decreased the intracellular ROS level statistically at combined doses, but 100μg/ml+1000 μM H2O2' It was observed that the extract at a combined dose of 1,0 increased the effect of H2O2 and increased the intracellular ROS level statistically significantly.
Cancer continues to be one of the leading causes of morbidity and mortality worldwide. Cancer is the second leading cause of death after cardiovascular diseases, excluding non-communicable diseases. Today, research on various herbal products is increasingly continuing to develop new drugs against diseases such as cancer. In recent years, promising anti-cancer efficacy of various herbal products against different cancers has been demonstrated. In this context, in the thesis study, it was aimed to investigate the anti-cancer potentials of methanolic extracts obtained from Anemone blanda plant. For this purpose, the human healthy bronchial epithelial cell line Beas-2B and the human carcinoma cell line A549 cells were used. XTT test and Clonogenic test were used to determine cytotoxicity, Komet method to determine DNA damages and ROS test to determine their effects on intracellular reactive oxygen level. As a result of the XTT test, the IC50 values in Beas-2B and A549 cell lines were determined as 21.90 μg/mL and 21.25 μg/mL, respectively. In the clonogenic test, these values were determined as 24.57 μg/mL and 27.62 μg/mL for Beas-2B and A549 cell lines, respectively. In the Comet test, it was used in XTT and Clonogenic tests, in addition to the doses, it was added in combined doses with H2O2. Tail length, tail % DNA and olive tail moment in Beas-2B and A549 cell lines were calculated, and it was determined that there were statistically significant increases in DNA strand breaks compared to the control group, while it was statistically decreased at doses with H2O2. While the same doses as in the comet test were used in the ROS test, there was no statistically significant change in the intra-ROS level for Beas-2B in the results. In combined doses, it was observed that the mean intracellular ROS level of Anemone extract was statistically significantly reduced from 40 μg/mL dose, while it statistically decreased the ROS level for A549, while it decreased the intracellular ROS level statistically at combined doses, but 100μg/ml+1000 μM H2O2' It was observed that the extract at a combined dose of 1,0 increased the effect of H2O2 and increased the intracellular ROS level statistically significantly.
Description
Keywords
Anemone blanda, Akciğer kanseri, Anti-kanser, A549, Beas-2B, Klononjenik testi, ROS testi, XTT testi, Lung cancer, Anti-cancer, Clonongenic test, Comet test, ROS test, XTT test
Citation
İrkören, S. F. (2022). Anemone blanda’nın sağlıklı (Beas-2B) ve kanser (A549) akciğer epitel hücre hatları üzerindeki in vitro antiproliferatif ve antioksidan etkilerinin araştırılması. Yayınlanmamış yüksek lisans tezi. Bursa Uludağ Üniversitesi Fen Bilimleri Enstitüsü.