Investigation of Apoptotic Effect of Klotho Protein on Human Colorectal Cancer Cells via TRAIL Death Receptors

Year 2022, Volume: 12 Issue: 2, 324 - 337, 30.12.2022

Sibel Gunes , Onur Uysal , Merve Nur Soykan , Ayla Eker Sarıboyacı

https://doi.org/10.37094/adyujsci.1212028

Abstract

Klotho is a transmembrane protein which is deficiency has pleiotropic effects in a number of aging-related disease processes and various cancers. In some recent studies about klotho protein disorders, it has been shown to klotho is effective in various cancers including lung, liver, breast, kidney, and colon. TNF-related apoptosis-inducing ligand (TRAIL), a TNF family molecule, is a cytokine that stimulates apoptosis through death receptors in many cancer types and therefore attracts attention for tumor therapies in various preclinical models. Interaction with TRAIL death receptors create an apoptotic effect in cancer treatments by reducing the proliferation of cancer cells. In this study, it was aimed to investigate the effects of exogen klotho administration on cell viability and apoptosis on TRAIL death receptors (TRAIL1 and TRAILR2) in human healthy colon cells (CCD 841 CoN) and TRAIL-resistant human colorectal cancer cells (caco2). For this purpose, cells were treated with different concentrations of klotho for 24 and 48 hours. To determine the cell viability, proliferation, and death receptors effects of klotho protein on cancer and healthy cells evaluations with WST-8 and Real-Time qRT-PCR analysis were performed. Our results showed that the increase in klotho protein concentration did not have a significant effect on cell viability in healthy colon cells, whereas it decreased cell viability and proliferation in apoptosis-resistant human colorectal cancer cells. Relative gene expression of TRAIL1 and TRAILR2 death receptors increased with klotho applied to the apoptosis-resistant colorectal cancer cell line caco-2. Therefore, targeting the ligand-receptors that induce TNF-related apoptosis with the klotho protein can be considered as a potential therapeutic approach for cancer therapy.

Keywords

Klotho, Cell viability, Colorectal cancer, Antitumor effect, TRAIL, Apoptosis

Supporting Institution

Eskişehir Osmangazi Üniversitesi

Project Number

ESOGU-BAP, Kod: 202046A113

Thanks

This study was supported by grants Eskisehir Osmangazi University, Scientific Research Projects (ESOGU-BAP, Code: 202046A113).

Klotho proteinin insan kolorektal kanser hücreleri üzerindeki apoptotik etkisinin TRAIL ölüm reseptörleri üzerinden incelenmesi

Abstract

Klotho, eksikliği bir dizi yaşlanma ile ilişkili hastalık sürecinde ve çeşitli kanserlerde pleiotropik etkilere sahip olan bir transmembran proteindir. Klotho protein bozuklukları ile ilgili son zamanlarda yapılan bazı araştırmalarda klothonun akciğer, karaciğer, meme, böbrek ve kolon dahil çeşitli kanserlerde etkili olduğu gösterilmiştir. Tümör nekroz faktör ilişkili apoptoz indükleyici ligand (TRAIL), birçok kanser türünde ölüm reseptörleri aracılığıyla apoptozu uyaran bir sitokindir ve bu nedenle çeşitli preklinik modellerde tümör terapileri için dikkat çekmektedir. TRAIL ölüm reseptörleri ile etkileşim, kanser hücrelerinin çoğalmasını azaltarak kanser tedavilerinde apoptotik bir etki yaratır. Bu çalışmada, eksojen klotho uygulamasının insan sağlıklı kolon (CCD 841 CoN) ve TRAIL dirençli insan kolorektal kanser hücreleri (Caco-2) üzerindeki hücre canlılığı ve apoptoz üzerine etkilerinin TRAIL ölüm reseptörleri (TRAIL1 ve TRAILR2) aracılığı ile araştırılması amaçlanmıştır. Bu amaçla hücreler, 24 ve 48 saat boyunca farklı konsantrasyonlarda klotho ile muamele edildi. Klotho proteinin kanser ve sağlıklı hücreler üzerindeki hücre canlılığı, proliferasyonu ve ölüm reseptörlerinin etkilerini belirlemek için WST-8 ve real-time qRT-PCR analizi ile değerlendirmeler yapıldı. Sonuçlarımız, klotho protein konsantrasyonundaki artışın sağlıklı kolon hücrelerinde hücre canlılığı üzerinde önemli bir etkiye sahip olmadığını, apoptoza dirençli insan kolorektal kanser hücrelerinde ise hücre canlılığını ve proliferasyonunu azalttığını gösterdi. Apoptoza dirençli kolorektal kanser hücre dizisi Caco-2'ye uygulanan klotho ile TRAIL1 ve TRAILR2 ölüm reseptörlerinin göreceli gen ekspresyonu arttı. Sonuç olarak, TRAIL ölüm reseptörlerinin klotho proteini ile hedeflenmesi kolorektal kanser tedavisi için potansiyel bir terapötik yaklaşım olarak kabul edilebilir.

Keywords

Klotho; Cell viability; Colorectal cancer; Antitumor effect; TRAIL, Apoptosis

Project Number

ESOGU-BAP, Kod: 202046A113

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