Combinational Effect of Methylene Blue-Mediated Photodynamic Therapy and Doxorubicin on Breast Cancer Cells

Öz

Objective: Breast cancer is the most common cancer in women and its incidence is increasing. Today, there are many methods in the treatment of breast cancer, such as surgery, radiation therapy and chemotherapy. Doxorubicin (DOX), one of the chemotherapeutics commonly used in chemotherapy treatment, causes dangerous side effects and drug resistance above a certain dose. Thus, it reduces the effect of the treatment. Compared to existing treatments, combination therapies have recently gained importance to overcome these problems. Therefore, this study aimed to reduce the dose of DOX used in breast cancer treatment and increase the effectiveness of the treatment by combining it with methylene blue (MB) activated by photodynamic therapy (PDT). Methods: Different concentrations of DOX, MB, MB-PDT, MB-PDT, DOX-PDT, MB-PDT+DOX and DOX+MB-PDT were tested separately and in combination on MCF-7 breast cancer cell line. MTT assay was performed to determine the cell viability and IC50 values were calculated where cells showed 50% viability. In addition, the combination index (CI) value was found to determine the degree of interaction between MB and DOX. In addition, mRNA expression levels of apoptotic and antiapoptotic markers (Bax, Bcl-2) were analysed by RT-PCR to determine the apoptotic potential of DOX and photodynamic therapy combination against breast cancer cells. Results: The combination of MB-PDT followed by low concentrations of DOX (428.4 nM) had a better effect in inducing cancer cell death compared to DOX alone and DOX followed by MB-PDT. At the same time, this combined treatment showed its apoptotic potential by decreasing Bcl-2 gene expression and increasing Bax gene expression. Conclusion: The combined treatment of doxorubicin and methylene blue-induced photodynamic therapy has shown synergistic, cytotoxic and apoptotic effects on breast cancer cells, offering a promising approach.

Anahtar Kelimeler

• Breast cancer
• doxorubicin
• photodynamic therapy
• methylene blue
• combination therapy

Meme Kanseri Hücresinde Metilen Mavisi Aracili Fotodinamik Terapi ve Doksorubisinin Kombinasyonel Etkisi

Öz

Öz Amaç: Meme kanseri kadınlarda görülen en sık kanser olup görülme sıklığı giderek artmaktadır. Günümüzde meme kanserinin tedavisinde cerrahi tedavi, radyasyon tedavisi ve kemoterapi gibi birçok yöntem vardır. Kemoterapi tedavisinde yaygın olarak kullanılan kemoterapötiklerden birisi olan doksorubisin (DOX) belirli bir dozun üzerinde tehlikeli yan etkilere ve ilaç direncine neden olur. Mevcut tedavilere kıyasla kombinasyon tedaviler son zamanlarda bu sorunların üstesinden gelmek için önem kazanmıştır. Bir fotoduyarlaştırıcı ile uygulanan fotodinamik terapi klinikte birçok kanser türünde uygulanmaktadır. Bu çalışma ile meme kanseri tedavisinde kullanılan DOX’u fotodinamik terapi (PDT) ile aktive edilen metilen mavisi (MB) ile kombinasyon uygulayarak DOX’un uygulanmasında düşük toksisite ile yüksek etkinlik alınması amaçlanmıştır. Yöntem: MCF-7 meme kanseri hücre hattı üzerinde farklı konsantrasyonlarda DOX, MB, MB-PDT, DOX-PDT, MB-PDT+DOX ve DOX+MB-PDT olacak şekilde ayrı ayrı ve kombinasyon tedaviler denenmiştir. Hücre canlılığını belirlemek için MTT testi gerçekleştirilmiş ve hücrelerin %50 canlılık gösterdiği IC50 değerleri hesaplanmıştır. Ayrıca, MB ve DOX arasındaki etkileşimin derecesini belirlemek için kombinasyon indeksi (CI) değeri bulunmuştur. Ayrıca DOX ve fotodinamik terapi kombinasyonunun meme kanseri hücrelerine karşı apoptotik potansiyelini belirlemek için apoptotik ve antiapoptotik belirteçlerin (Bax, Bcl-2) mRNA ekspresyon seviyelerine RT-PCR ile bakılmıştır. Bulgular: MB-PDT ve ardından düşük konsnatrasyonda DOX (428.4 nM ) kombinasyonunun, tek başına DOX'a ve önce DOX ardından MB-PDT’ye kıyasla kanser hücresi ölümünü indüklemede daha iyi bir etkiye sahip olduğunu göstermiştir. Aynı zamanda bu kombine uygulama Bcl-2 gen ekspresyonunu düşürürken, Bax gen ekspresyonunu yükselterek apoptotik potansiyelini göstermiştir. Sonuç: Doksorubisin ve metilen mavisi ile indüklenmiş fotodinamik terapi kombine tedavisi meme kanseri hücreleri üzerinde sinerjik, sitotoksik ve apoptotik etkiler göstererek , umut verici bir yaklaşım sunmuştur.

Anahtar Kelimeler

• Meme kanseri
• doksorubisin
• fotodinamik terapi
• metilen mavisi
• kombinasyon tedavi.

Kaynakça

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