An Investigation of Anticancer Effects of Doxorubicin and Calcitriol Combination on MCF-7 Cells

Abstract

DOI: 10.26650/experimed.2018.18001

Objectives: This study aimed to identify a substance that both increases the efficiency and decreases the dose of doxorubicin. Doxorubicin has considerable cardiac side effects at certain doses when used treating breast cancer. Calcitriol, one of the vitamin D analogs considered to have antiproliferative effects, was selected, and its cytotoxic effects on the human breast cancer cell line MCF-7 were investigated in combination with doxorubicin.

Materials and Methods: MCF-7 cell line was treated with calcitriol in real time for 72 h in x-CELLigence. The antiproliferative optimal dose of calcitriol was determined by time-dependent cell index graph plotted using The xCELLigence Real-Time Cell Analysis (RTCA) software program. The combination of different doses of doxorubicin and this optimal dose of calcitriol was used to treat the MCF-7 cell line. Then, Sulforhodamine-B (SRB) assay was conducted, and spectrophotometric measurements were performed for cytotoxicity assay. The results of these spectrophotometric measurements were analyzed by Student’s -test.

Results: The optimal antiproliferative calcitriol dose detection of MCF-7 cells was performed using the time-dependent cell index graph RTCA software program. Spectrophotometric measurements obtained using the protein-staining sulforodamine B (SRB) assay for cytotoxicity determination were statistically evaluated by the Student’s t-test using the GraphPad Prism program. The optimal dose of calcitriol was determined to be 250 nM. Different doses of doxorubicin (1.84-0.92 µM), calcitriol (250 nM), and calcitriol without the MCF-7 cell line were then used for detecting the cytotoxic effect. The combination of 0.46 µM doxorubicin and the optimal dose of calcitriol was found to be cytotoxic compared with other doses (p=0.0087); however, it was not as effective as the dose reduction obtained when using doxorubicin.

Conclusion: The combined use of doxorubicin with calcitriol was found to have no significant effect in reducing the doses presently being used. Hence, it is too early to state that a combination of vitamin D and doxorubicin in breast cancer treatment will not have any beneficial effects. Other vitamin D analogs might be potential candidates for breast cancer treatment in further studies.

Keywords

• Breast cancer,doxorubicin,calcitriol,MCF-7,cell culture

Kalsitriol ve Doksorubisin Kombinasyonunun MCF-7 Üzerine Olası Anti Kanser Etkilerinin Araştırılması

Abstract

DOI: 10.26650/experimed.2018.18001

Amaç: Bu çalışmada amacımız, belirli bir dozun üzerinde kardiyak yan etkileri oldukça fazla olan ve meme kanseri tedavisinde kullanılan kemoterapötiklerden doksorubisinin, tedavide kullanım miktarlarını azaltmayı ve etkinliğini arttırmayı sağlayan bir maddeyi saptamaktı. Etken madde olarak anti proliferatif etkisi olduğu düşünülen vitamin D analoglarından biri olan kalsitriol seçilmiş ve doksorubisin ile kombine tedavisinin, insan meme kanseri hücre hattı MCF-7 üzerine sitotoksik etkisinin saptanması amaçlanmıştır.

Gereç ve Yöntem: MCF-7 insan meme kanseri hücre hattı kalsitriol ile muamele edilerek, gerçek zamanlı olarak, x-CELLigence cihazında 72 saat inkübasyona bırakıldı ve kalsitriolün anti-proliferatif optimum doz tespiti zamana bağlı hücre indeksi grafiği The xCELLigence Real-Time Cell Analysis (RTCA) software programı kullanılarak yapıldı. Kalsitriol optimum dozu ve doksorubisinin farklı dozlarının kombinasyonu MCF-7 hücre kültürü ile muamele edilerek sitotoksisite tayini için Sulforhodamine-B (SRB) uygulaması ve spektrofotometrik ölçüm uygulandı. Spektrofotometrik ölçüm sonuçları anlamlılık sonuçları için Student’s t-Testi ile değerlendirildi.

Bulgular: MCF-7 hücrelerinin antiproliferative optimum kalsitriol doz tespiti zamana bağlı hücre indeksi grafiği RTCA software programı kullanılarak yapılmıştır. Sitotoksisite tayini için uygulanan SRB yöntemi sonucu elde edilen spektrofotometrik ölçüm sonuçları GraphPad Prism programı kullanılarak Student’s t-testi ile istatistiksel olarak değerlendirilmiştir. Kalsitriol optimum dozu 250 nM tespit edilmiştir. Doksorubisinin farklı dozları (1,84 µM-0,92 µM), kalsitriol (250 nM) ve kalsitriol olmaksızın MCF-7 hücre hattı ile sitotoksik etki saptaması için muamele edilmiştir. 0,46 µM doksorubisin ve optimum kalsitriol kombinasyonunun sitotoksik açıdan diğer dozlara göre anlamlı olduğu (p=0,0087) fakat doksorubisin kullanımında doz azaltımını sağlayacak kadar etkin olmadığı saptanmıştır.

Sonuç: Doksorubisinin, kalsitriol ile kombine kullanımının günümüzde kullanılan dozlar üzerinde azaltıcı yönde anlamlı bir etkisinin olmadığını saptanmıştır. Vitamin D ve doksorubisin birlikte kullanımının meme kanserinde fayda sağlamayacağını söylemek için ise henüz erkendir. İlerleyen çalışmalarda vitamin D’nin farklı analogları ile çeşitli çalışmalar yapılabilir.

Keywords

• Meme Kanseri,doksorubisin,kalsitriol,MCF-7,hücre kültürü

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