DNA BINDING ACTIVITIES OF THE VINCA ALKALOIDS AND PACLITAXEL AS ANTI-MICROTUBULE DRUGS USED IN CANCER THERAPY

Year 2022, , 51 - 57, 17.01.2022

Emine Öksüzoğlu

https://doi.org/10.18229/kocatepetip.883250

Abstract

OBJECTIVE: Microtubules are an essential part of the intracellular cytoskeletal structure and possess unique polymerization dynamics that are critical for many cellular functions, including cell division. Anti-microtubule drugs that interfere with microtubule formation are important chemotherapeutic agents for the treatment of various cancer. These drugs that block mitosis seem to work by a common mechanism, which suppresses the dynamics of microtubules, slows cells, induces apoptosis and subsequently kills tumor cells. Vinca alkaloids (vinblastine, vincristine and vinorelbine) and Taxanes (paclitaxel) are two different classes of anti-microtubule drugs that cause microtubule dysfunction and inhibit cancer cell proliferation. The main activity of vinca alkaloids and taxanes result from their binding interactions with tubulin proteins. However, studies on DNA interactions of these anti-microtubule drugs are not sufficient. In this study, it was aimed to investigate the DNA binding activities of the vinca alkaloids (vinblastine, vincristine, vinorelbine) and paclitaxel.
MATERIAL AND METHODS: The interactions of the drugs with DNA were analyzed by agarose gel electrophoresis assay. Three types of DNA were used in each experiment, including 100bp marker DNA, pUC19 plasmid DNA (2686 bp), and pBR322 plasmid DNA (4361 bp). After the DNAs were incubated with different concentrations of the drugs under certain conditions, agarose gel electrophoresis was performed. DNA band distributions were analyzed with a gel analysis system so that the drugs-DNA interactions could be interpreted.
RESULTS: According to our results, it was found that among the vinca alkaloids, especially vinorelbine binds to DNA with higher activity than vincristine and vinblastine. The Vinca alkaloids have structural properties required for DNA binding activity and there is a similarity in their DNA binding models. However, the results showed that paclitaxel, which is from the taxane group, did not have DNA binding activity. This may be because the chemical structure of paclitaxel is not suitable for binding to DNA.
CONCLUSIONS: The interaction of drugs with DNA play an important role in determining the pathways of drugs action and their ability to cause DNA damage. Consequently, the findings of our study will contribute to elucidating the effect mechanisms and the genotoxic potentials of these drugs, which are microtubule inhibitors.

Keywords

Anti-microtubule drugs, Vinca Alkaloids, Paclitaxel, DNA-binding, Cancer Therapy

KANSER TEDAVİSİNDE KULLANILAN ANTİ-MİKROTÜBÜL İLAÇLARDAN VİNCA ALKALOİDLER VE PAKLİTAKSEL’İN DNA BAĞLANMA AKTİVİTELERİ

Abstract

AMAÇ: Mikrotübüller, hücre içi hücre iskeleti yapısının önemli bir parçasıdır ve hücre bölünmesi dahil birçok hücresel işlev için kritik olan benzersiz polimerizasyon dinamiklerine sahiptirler. Mikrotübüllere müdahale eden anti-mikrotübül ilaçlar, çeşitli kanserlerin tedavisi için çok önemli kemoterapötik ajanlardır. Mitozu bloke eden bu ilaçlar, mikrotübüllerin dinamiğini baskılayan, hücreleri yavaşlatan, apoptozu indükleyen ve ardından tümör hücrelerini öldüren ortak bir mekanizma ile çalışmaktadır. Vinka alkaloidleri (vinblastin, vincristin ve vinorelbin) ve Taksanlar (paklitaksel), mikrotübüllerin işlev bozukluğuna neden olan ve kanser hücresi proliferasyonunu inhibe eden iki farklı anti-mikrotübül ilaç sınıfıdır. Vinka alkaloidlerinin ve taksanların ana aktivitesi, tübülin proteinleri ile bağlanma etkileşimlerinden kaynaklanmaktadır. Ancak, bu anti-mikrotübül ilaçların DNA etkileşimleri ile ilgili çalışmalar yeterli değildir. Bu çalışmada, vinka alkaloidleri (vinblastin, vincristin, vinorelbin) ve paklitakselin DNA bağlanma aktivitelerinin araştırılması amaçlanmıştır.
GEREÇ VE YÖNTEM: İlaçların DNA ile etkileşimleri agaroz jel elektroforez deneyi ile analiz edildi. Her deneyde, 100 bp marker DNA, pUC19 plazmid DNA (2686 bp) ve pBR322 plazmid DNA (4361 bp) dahil olmak üzere üç tip DNA kullanıldı. DNA'lar belirli koşullar altında farklı ilaç konsantrasyonları ile inkübe edildikten sonra agaroz jel elektroforezi yapıldı. İlaç-DNA etkileşimlerinin yorumlanabilmesi için DNA bant dağılımları jel analiz sistemi ile analiz edildi.
BULGULAR: Sonuçlarımıza göre vinka alkaloidlerinden özellikle vinorelbinin, vinkristin ve vinblastine göre daha yüksek aktiviteyle DNA'ya bağlandığı bulunmuştur. Vinca alkaloidleri, DNA bağlanma aktivitesi için gerekli yapısal özelliklere sahiptir ve DNA bağlanma motiflerinde benzerlik vardır. Ancak sonuçlar, taksan grubundan olan paklitakselin DNA bağlama aktivitesine sahip olmadığını gösterdi. Bunun nedeni, paklitakselin kimyasal yapısının DNA'ya bağlanmaya uygun olmaması olabilir.
SONUÇ: İlaçların DNA ile interaksiyonu, ilaçların etki yollarını ve DNA hasarına neden olma yeteneklerini belirlemede önemli bir rol oynar. Sonuç olarak, çalışmamızın bulguları mikrotübül inhibitörü olan bu ilaçların etki mekanizmalarını ve genotoksik potansiyellerini aydınlatmaya katkı sağlayacaktır.

Keywords

Anti-mikrotübül ilaçlar, Vinca alkaloidleri, Paklitaksel, DNA-bağlanma, Kanser terapi

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