Anticancer and Apoptotic Effects of Nor-wogonin on Human Cervical Cancer Cells

Abstract

Cancer, a growing health problem all over the world, is the second most important cause of death after cardiovascular diseases. The ineffectiveness of classical methods (surgery, radiotherapy, chemotherapy, etc.) used in cancer treatment has led researchers to discover and develop new therapeutics in cancer treatment. It is known that flavones, a subgroup of flavonoids, have anticancer, anti-inflammatory, antiprolerative, and antiangiogenic effects in addition to their biochemical and pharmacological effects. We aimed to determine the cytotoxic effect of Nor-wogonin (5, 7, 8-trihydroxyflavone), a polyhydroxyflavone derivative with various biological activities, on the human cervical cancer cell line and its effect on the expression level of some genes involved in apoptosis. In the study, different concentrations of Nor-wogonin (10, 20, 40, 80 µM) were applied to the cervical (HeLa) cancer cell line. The cytotoxic effect of the compound on cell lines was determined by the MTT (3-(4,5-dimethylthiazol-2-yl)-diphenyl tetrazolium bromide) method and the Nor-wogonin IC50 value for HeLa cells was calculated. In addition, quantitative gene analysis by Q-PCR was performed to observe the effect of Nor-wogonin on the levels of anti-apoptotic and pro-apoptotic gene expression (bax, p53, caspase-3 and bcl-2 genes) in HeLa cells. It was determined that Nor-wogonin significantly decreased cell viability in HeLa cells depending on the increasing dose (p<0.05). IC50 values for HeLa cells were calculated as 32.09 µM. In addition, it was determined that bax, caspase-3, p53 gene expressions increased and bcl-2 gene expression decreased in HeLa cells. In conclusion, Nor-wogonin, a flavone derivative, can be considered as an important therapeutic agent thanks to its in vitro cytotoxic activity and its effects on anti-apoptotic and pro-apoptotic gene expression.

Keywords

• Nor-wogonin
• Cervical cancer
• Antikanser effect
• Apoptotic effect
• Cell culture.

Nor-wogonin’in İnsan Rahim Ağzı Kanseri Hücrelerinde Antikanser ve Apoptotik Etkileri

Öz

Tüm dünyada büyüyen bir sağlık sorunu olan kanser, kardiyovasküler hastalıklardan sonra ikinci en önemli ölüm nedenidir. Kanser tedavisinde kullanılan klasik yöntemlerin (cerrahi, radyoterapi, kemoterapi vb.) tedavide istenilen düzeyde etkili olamaması araştırmacıları kanser tedavisinde yeni terapötiklerin keşfine ve geliştirilmesine yönlendirmiştir. Flavonoidlerin bir alt grubu olan flavonların biyokimyasal ve farmakolojik etkilerine ek olarak antikanser, antienflamatuvar, antiproleratif, antianjiyogenik etkileri olduğu bilinmektedir. Çeşitli biyolojik aktivitelere sahip bir polihidroksi flavon türevi olan Nor-wogonin'in (5, 7, 8-trihydroxyflavone) insan rahim ağzı kanseri hücre hattı üzerine sitotoksik etkisini ve apoptoza dahil olan bazı genlerin eksprosyon düzeyine etkisini belirlemeyi amaçladık. Çalışmada, rahim ağzı (HeLa) kanser hücre hattına farklı konsantrasyonlarda (10, 20, 40, 80 µM) Nor-wogonin uygulandı. Bileşiğin hücre hatları üzerindeki sitotoksik etkisi MTT (3-(4,5-dimetiltiyazol-2-il)-difenil tetrazolyum bromür) yöntemi ile belirlendi ve HeLa hücreleri için Nor-wogonin IC50 değeri hesaplandı. Ek olarak, Nor-wogonin'in HeLa hücrelerindeki anti-apoptotik ve pro-apoptotik gen ekspresyon (bax, p53, caspase-3 ve bcl-2 genleri) seviyeleri üzerindeki etkisini gözlemlemek için Q-PCR ile kantitatif gen analizi yapıldı. Nor-wogonin'in artan doza bağlı olarak HeLa hücrelerinde hücre canlılığını önemli ölçüde azalttığı belirlendi (p<0.05). HeLa hücreleri için IC50 değerleri 32.09 µM olarak hesaplandı. Ayrıca HeLa hücrelerinde bax, kaspaz-3, p53 gen ekspresyonlarının arttığı ve bcl-2 gen ekspresyonunun azaldığı belirlendi. Sonuç olarak, bir flavon türevi olan Nor-wogonin, in vitro sitotoksik aktivitesi ile anti-apoptotik ve pro-apoptotik gen ekspresyonu üzerindeki etkileri sayesinde önemli bir terapötik ajan olarak kabul edilebilir.

Anahtar Kelimeler

• Nor-wogonin
• Rahim ağzı kanseri
• Antikanser etki
• Apoptotik Etki
• Hücre kültürü.

Kaynakça

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