Kolşisin’in MCF-7 insan meme adenokarsinoma hücrelerinde hücre döngüsü tutulumu ve MMP-2 mRNA ifade seviyesi üzerine etkisi

Öz

Amaç: Kolşisin, trisiklik alkaloid bir ilaç olup klinikte anti-enflamatuvar etkisinden dolayı yaygın olarak kullanılmaktadır. Kolşisin uygun konsantrasyonlarda oral yolla kullanılmakta iken yüksek konsantrasyonlarda toksik etki gösterdiği tespit edilmiştir. Bu nedenle, kanser araştırmalarında düşük doz kolşisinin etki mekanizmaları üzerine çalışmalar yapılmaktadır. Matriks metalloproteinaz-2 MMP-2 , matriks metalloproteinaz enzim ailesinin bir üyesi olup kanserde ifade seviyesinin arttığı ve ekstrasellüler matriksi yıkıma uğratarak kanser hücrelerinin metastazına neden olduğu bildirilmiştir. MMP’lerin bu önemli etkisi nedeni ile kanser hastalığında metastazın önlenmesi amacıyla çeşitli bileşiklerin MMP üzerine inhibitör etkisinin değerlendirilmesi kanser araştırmalarında önemli yer tutmaktadır. Bu çalışmada, kolşisinin, MCF-7 insan meme adenokarsinoma hücresinde hücre döngüsü tutulumu ve metastazda önemli rol oynadığı bilinen MMP-2 protein ifadesi üzerine etkisinin araştırılması amaçlanmıştır. Yöntem: Çalışmada, MCF-7 insan meme adenokarsinoma hücreleri kullanılmıştır ATCC, HTB22 . Hücrelere 0.1, 10 ve 100 μg/ml konsantrasyonlarda kolşisin uygulanmış ve hücre canlılığı analizi MTT testi ile yapılmıştır. Kolşisin’in hücre döngüsü tutulumu üzerine etkisi analiz edilmiş ve G0/G1, S ve G2/M döngüsündeki hücre populasyonları yüzde % olarak ifade edilmiştir. Kolşisin’in MMP-2 mRNA ifade seviyesi üzerine etkisi ise gerçek zamanlı PCR qRT-PCR yöntemi ile tayin edilmiştir. Bulgular: 10 ve 100 μg/ml konsantrasyonlarda Kolşisin MCF-7 meme kanser hücresinin canlılığını istatistiksel olarak anlamlı derecede inhibe etmiştir. Kolşisin’in tüm konsantrasyonlarda hücre döngüsünü G2/M fazında inhibe ettiği ve MMP-2 geninin ifade seviyesini anlamlı olarak azalttığı saptanmıştır p

Anahtar Kelimeler

• kolşisin, hücre döngüsü, MMP-2 ekspresyonu, antikanser, MCF-7

Effects of colchicine on cell cycle arrest and MMP-2 mRNA expression in MCF-7 breast adenocarcinoma cells

Abstract

Objective: Colchicine is a tricyclic alkaloid drug and it’s been clinically used for a long time because of its anti-inflammatory effects. While colchicine has been safely used in oral applications, it’s been demonstrated that colchicine is toxic at higher concentrations. So, the mechanism of effects of low dose colchicine has been studied in cancer related researches. Matrix metalloproteinase-2 MMP-2 is a member of matrix metalloproteinase enzyme family and it’s been demonstrated that MMP-2 expression is increased in cancer and it causes a metastasis of cancer cells through degradation of extracellular matrix. Because of this important effect of MMPs, the studies related to develop MMP inhibitor compounds have great importance in cancer investigations. At present study, it’s aimed to investigate the effects of colchicine on cell cycle arrest and MMP-2 mRNA expression in MCF-7 human breast cancer cells. Methods: In the study, MCF-7 human breast adenocarcinoma cells were purchased from ATCC. Cells were treated with 0.1, 10 ve 100 μg/ml colchicine and cell viability was determined via MTT assay. The effect of colchicine on cell cycle arrest was determined by Muse Cell Analyzer and the percent of cell populations at G0/G1, S and G2/M cycles were identified. The effect of colchicine on MMP-2 mRNA expression has been performed by real-time PCR qRTPCR analysis. Results: Colchicine has significantly inhibited cell viability at 10 and 100 μg/ml concentrations. It’s been also demonstrated that colchicine has induced a cell cycle arrest at G2/M phase and downregulated MMP-2 mRNA expression of MCF-7 cells in all treated concentration p

Keywords

• colchicine, cell cycle, MMP-2 expression, anticancer, MCF-7

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