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

Year 2018, Volume: 8 Issue: 2, 41 - 46, 12.11.2018

Özge Bildiren Buse Cevatemre Ebru Nur Ay Güneş Özen Ceylan Hepokur Merve Erkısa Özlem Küçükhüseyin Engin Ulukaya Serap Kuruca İlhan Yaylım

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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