Comparison of Apoptotic Effects of Rapamycin and Vemurafenib in Colorectal Cancer

Abstract

The third most prevalent cancer in the world is colorectal cancer (CRC). Chemotherapy and surgery are the primary CRC treatments. Patients with CRC develop resistance to the chemotherapy treatments, which leads to the chemotherapy's failure. Due to this, various molecular processes that contribute to the development and progression of CRC have come into attention recently. mTOR and MAPK signaling pathways are crucial for the growth of CRC and mTOR pathway inhibitor RAPA and the MAPK pathway inhibitor VMF affected the apoptosis of CRC cells. Human CRC cells HT29 were cultured. The WST-1 test was used to identify the optimum dose of RAPA and VMF on HT29 CRC cells. To determine the apoptotic effects of RAPA and VMF on HT29 cells, flow cytometry was performed. Statistical significance level was accepted as p≤0.05. According to the data we obtained, the dose of RAPA and VMF to be applied to HT29 cells was found to be 46.97 μM and 35.84 μM, respectively, at the 24th hour. RAPA was found to have a greater impact on apoptosis in HT29 cells than VMF (p<0.0001 for RAPA, p<0.01 for VMF). In summary, our study found that RAPA and VMF decreased the viability of HT29 colorectal cancer cells and caused apoptosis via the caspase 3/7 pathway.

Keywords

• Colorectal cancer
• mTOR
• MAPK
• Rapamycin
• Vemurafenib

Kolorektal Kanserde Rapamisin ve Vemurafenib’in Apoptotik Etkilerinin Karşılaştırılması

Abstract

Kolorektal kanser (KRK), dünyada en sık görülen üçüncü kanser türüdür. KRK’de ilk tedavi seçeneği cerrahi ve kemoterapidir. Ancak, kullanılan ilaçlara karşı gelişen direnç, uygulanan kemoterapinin başarısız olmasına yol açmaktadır. Son yıllarda yeni teröpatik ajan arayışları, KRK gelişimi ve ilerlemesinde rol oynayan farklı moleküler mekanizmalar üzerinde yoğunlaşmaktadır. mTOR ve MAPK sinyal yolaklarının KRK gelişiminde anahtar rol oynadığı bilinmektedir. Bu çalışmada, KRK hücrelerinde mTOR yolağı inhibitörü Rapamisin (RAPA) ve MAPK yolağı inhibitörü Vemurafenib (VMF)’in apoptoz üzerine olan etkilerinin karşılaştırılması amaçlanmıştır. Çalışmamızda, insan KRK hücrelerinde üretilen HT29 hücre hattı kültüre edilmiştir. RAPA ve VMF’nin HT29 KRK hücreleri üzerindeki uygun dozunun belirlenmesi için WST-1 testi ve apoptotik etkilerinin belirlenmesi amacıyla da akım sitometrisi yöntemi kullanılmıştır. İstatistiksel anlamlılık düzeyi p≤0.05 olarak kabul edilmiştir. Elde ettiğimiz verilere göre, HT29 hücrelerine uygulanacak olan RAPA ve VMF dozu 24. saatte sırasıyla 46,97 μM ve 35,84 μM olarak bulunmuştur. HT29 hücrelerinde RAPA’nın apoptotik süreç üzerinde VMF’den daha etkin olduğu bulunmuştur (RAPA için; p<0.0001, VMF için; p<0.01). Sonuç olarak, çalışmamızda RAPA ve VMF’nin HT29 kolorektal kanser hücrelerinin canlılığını azalttığı ve apoptozu kaspaz 3/7 yolu ile indüklediği görülmüştür.

Keywords

• Kolorektal kanser
• mTOR
• MAPK
• Rapamisin
• Vemurafenib

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