Mitochondrial Transfer Drives Stem Cell–Like Phenotypes in Gastric Cancer Cells

Öz

Background and Objective: Mitochondrial dynamics have a crucial role in the development of cancer and the plasticity of cancer cells. The role of mitochondria isolated from parental cells in regulating mitochondrial content and cancer stem cell (CSC) properties of MKN45 gastric cancer cells was examined. Methods: Mitochondria were isolated from parental MKN45 cells and transferred into recipient cells using a co-incubation approach. Mitochondrial DNA (mtDNA) copy number was analyzed by quantitative real-time PCR (qPCR). Fluorescence microscopy was used to confirm the presence of mitochondria stained with Rhodamine 123 in the recipient cells following the transfer of mitochondria. Self-renewal capacity was evaluated using a sphere formation assay. The expression levels of stemness (OCT4, NANOG, BMI1), mesenchymal and epithelial (SNAIL, SLUG, VIMENTIN, E-CADHERIN) markers were determined by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Results: The transfer of mitochondria caused a significant increase in the number of mtDNA copies and a higher mitochondrial membrane potential in the recipient cells. Furthermore, these mitochondrial alterations were functionally associated with an increase in the ability to form spheres and the expression of genes related to stemness, mesenchymal marker expression, and epithelial marker expression. Conclusion: These results suggest that the augmented mitochondrial content enhances the CSC-like traits that could be a factor in tumor aggressiveness.

Anahtar Kelimeler

• mitochondria
• mitochondria isolation
• mitochondria transfer
• mitochondrial DNA
• gastric cancer

Mitokondriyal Transferin Gastrik Kanser Hücrelerinde Kök Hücre Benzeri Özelliklerin Ortaya Çıkışındaki Rolü

Öz

Amaç: Mitokondriyal dinamikler, kanserin gelişiminde ve kanser hücrelerinin plastisitesinde kritik bir rol oynamaktadır. Bu çalışmada, ebeveyn hücrelerden izole edilen mitokondrilerin MKN45 mide kanseri hücrelerinin mitokondriyal içeriği ve kanser kök hücresi (CSC) özellikleri üzerindeki rolü incelenmiştir. Yöntemler: Mitokondriler, ebeveyn MKN45 hücrelerinden izole edilerek alıcı hücrelere birlikte inkübasyon yöntemiyle aktarıldı. Mitokondriyal DNA (mtDNA) kopya sayısı eş zamanlı qPCR ile analiz edildi. Mitokondri transferinden sonra alıcı hücrelerde Rhodamine 123 ile boyanmış mitokondri varlığını doğrulamak için floresan mikroskopi kullanıldı. Öz yenilenme kapasitesini değerlendirmek için küre (sphere) oluşum analizi yapıldı ve kök hücre belirteçleri (OCT4, NANOG, BMI1) ile mezenkimal ve epitelyal belirteçlerin (SNAIL, SLUG, VIMENTIN, E-KADERİN) gen ekspresyon düzeyleri RT-qPCR ile belirlendi. Bulgular: Mitokondrilerin aktarımı, alıcı hücrelerde mtDNA kopya sayısında dikkat çekici bir artışa ve daha yüksek bir mitokondriyal membran potansiyeline neden olmuştur. Ayrıca, bu mitokondriyal değişikliklerin, küre (sphere) oluşturma yeteneğinde ve kök hücreliliğe (stemness) ilişkin genlerin (OCT4, NANOG, BMI1) yanı sıra mezenkimal ve epitelyal belirteçlerin (SNAIL, SLUG, VIMENTIN, E-KADERİN) ekspresyonunda artışla işlevsel olarak bağlantılı olduğu görülmüştür. Sonuç: Bu sonuçlar, artan mitokondriyal içeriğin CSC özelliklerini güçlendirdiğini ve bunun tümör agresifliğinde etkili bir faktör olabileceğini düşündürmektedir.

Anahtar Kelimeler

• mitokondri
• mitokondri izolasyonu
• mitokondri transferi
• mitokondriyal DNA
• mide kanseri

Kaynakça

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