HDACs modulation and anticancer potential of Euphorbia virgata in colon adenocarcinoma: experimental and bioinformatics insights

Abstract

Aim: Euphorbia virgata (E. virgata), a species belonging to the Euphorbiaceae family, is distributed across diverse regions of the world. Owing to its rich phytochemical profile, particularly phenolics and alkaloids, it exhibits a wide range of biological activities. This study investigated the effects of ethanol extract from the aerial parts of E. virgata on HT-29 COAD and CCD-18Co normal fibroblast cell lines, as well as the expression profiles of HDACs. Material and Methods: The plant material utilised in the present study was collected from the village of İmaret, located in Sivas province, during the months of June and July 2023. Prof. Dr. Yavuz Bülent Köse subsequently identified the specimen, which was then deposited at the Anadolu University Herbarium under specimen number 16195. Ethanol extracts from the aerial parts of E. virgata were applied to HT-29 and CCD-18Co cell lines at concentrations ranging from 0.5 to 250 µg/mL. Anticancer activity was assessed using the MTT assay. The expression levels of HDACs were analysed by RT-PCR using the ∆∆Ct method. Results: Based on the cell culture experiments, anticancer activity was evaluated using the MTT assay. The IC₅₀ values for CCD-18Co cells were 149.9 µg/mL at 24 h, 36.37 µg/mL at 48 h, and 15.98 µg/mL at 72 h. In contrast, the IC₅₀ values for HT-29 cells were 3.9 µg/mL at 24 h, 1.295 µg/mL at 48 h, and 0.4653 µg/mL at 72 h. The expression levels of HDACs were determined in HT-29 COAD cells in comparison to CCD-18Co healthy control cells. The results demonstrated that the isolated E. virgata extract exhibited increased HDAC1 and HDAC3 gene expression. Furthermore, HDAC2 expression was decreased, demonstrating a significant inhibitory effect on E. virgata. Conclusions: In conclusion, E. virgata may play an important role in HDAC regulation in COAD cells and may offer a potential therapeutic option for preventing COAD progression. However, further studies are required to confirm these findings and assess their transferability to clinical practice.

Keywords

• E. virgata
• COAD
• HDAC1
• HDAC2
• HDAC3

Kolon adenokarsinomunda Euphorbia virgata'nın HDAC'ların modülasyonu ve antikanser potansiyeli: deneysel ve biyoenformatik bakış açıları

Abstract

Amaç: Euphorbiaceae familyasına ait bir tür olan Euphorbia virgata (E. virgata), dünyanın çeşitli bölgelerine yayılmıştır. Özellikle fenolikler ve alkaloidler olmak üzere zengin fitokimyasal profili sayesinde geniş bir biyolojik aktivite yelpazesi sergiler. Bu çalışmada, E. virgata'nın toprak üstü kısımlarından elde edilen etanol özütünün HT-29 COAD ve CCD-18Co normal fibroblast hücre hatları üzerindeki etkileri ve HDAC'lerin ekspresyon profilleri araştırılmıştır. Gereç ve Yöntemler: Çalışmada kullanılan bitki materyali, Haziran ve Temmuz 2023 aylarında Sivas ilinin İmaret köyünden toplanmıştır. Prof. Dr. Yavuz Bülent Köse tarafından teşhis edilen örnek, Anadolu Üniversitesi Herbaryumu'na 16195 numaralı örnekle teslim edilmiştir. E. virgata'nın toprak üstü kısımlarından elde edilen etanol ekstraktları, HT-29 ve CCD-18Co hücre hatlarına 0,5 ila 250 µg/mL arasında değişen konsantrasyonlarda uygulanmıştır. Antikanser aktivitesi MTT yöntemi kullanılarak değerlendirilmiştir. HDAC'lerin ekspresyon düzeyleri, ∆∆Ct yöntemi kullanılarak RT-PCR ile analiz edilmiştir. Sonuç: Hücre kültürü deneylerine dayanarak, antikanser aktivitesi MTT testi kullanılarak değerlendirildi. CCD-18Co hücreleri için IC₅₀ değerleri 24 saatte 149,9 µg/mL, 48 saatte 36,37 µg/mL ve 72 saatte 15,98 µg/mL olarak bulundu. Buna karşılık, HT-29 hücreleri için IC₅₀ değerleri 24 saatte 3,9 µg/mL, 48 saatte 1,295 µg/mL ve 72 saatte 0,4653 µg/mL olarak bulundu. HDAC'lerin ifade seviyeleri, HT-29 COAD hücrelerinde CCD-18Co sağlıklı kontrol hücreleriyle karşılaştırıldığında belirlendi. Sonuçlar, izole edilmiş E. virgata özütünün HDAC1 ve HDAC3 gen ifadesinde artış gösterdiğini gösterdi. Ayrıca, HDAC2 ekspresyonunun azaldığı ve E. virgata üzerinde önemli bir inhibitör etki gösterdiği görüldü.

Keywords

• E. virgata
• COAD
• HDAC1
• HDAC2
• HDAC3

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