Determination of Inhibitory Effects of Newly Synthesized Potential HIF Inhibitors on Non-Small Cell Lung Cancer Under Hypoxic Conditions

Yıl 2023, Cilt: 10 Sayı: 3, 177 - 182, 19.12.2023

Demet Kahraman , Pınar Yumrutaş , Esra Bozgeyik , İbrahim Bozgeyik , Ayşegül İyidoğan , Emine Elçin Oruç-emre , Serdar Öztuzcu , Ahmet Ferudun Işık

https://doi.org/10.47572/muskutd.1329302

Öz

According to global estimates, there are 2.3 million new cases and 1.8 million fatalities due to lung cancer each year. Despite recent progress in diagnosis and treatment, persistent challenges highlight the urgent need for novel therapeutics and innovative approaches to combat lung cancer effectively. Accordingly, in the present study, we aimed to investigate the anticancer properties of potential inhibitors of HIF-1α, compound 7a and 7b. In the study, HTB-54 and BEAS-2B cell lines were used. MTT cell viability experiments were performed to determine the effect of newly synthesized HIF inhibitors 7a and 7b on cell viability under normoxic and hypoxic conditions. Quantitative expression levels of HIF1A were determined by real-time PCR approach. While the half maximum inhibitory concentration (IC50) of compound 7a in HTB-54 cells was 10.37 µM under normoxic conditions, it was found to be 10.63 µM under hypoxic conditions. The IC50 value of another HIF inhibitor 7b in HTB-54 cells was found to be 8.80 µM under normoxic conditions and 9.54 µM under hypoxic conditions. The expression level of HIF1A was found to be lower in cells exposed to compounds 7a and 7b under hypoxia compared to the control group. Conversely, in normoxia, HIF1A expression level in cells exposed to compound 7a increased 6.5-fold (p<0.0001) compared to the control group, while it was found to increase approximately 9-fold (p<0.0001) when exposed to 7b. Consequently, both compound 7a and 7b holds great promise for future therapeutic interventions to lung cancer.

Anahtar Kelimeler

Lung Cancer, HIF Inhibitors, Hypoxia

Yeni Sentezlenen Potansiyel HIF İnhibitörlerinin Hipoksik Koşullarda Küçük Hücre Dışı Akciğer Kanseri Üzerindeki İnhibitör Etkilerinin Belirlenmesi

Öz

Küresel tahminlere göre her yıl akciğer kanseri nedeniyle 2,3 milyon yeni vaka ve 1,8 milyon ölüm yaşanmaktadır. Tanı ve tedavideki son gelişmelere rağmen devam eden zorluklar, akciğer kanseriyle etkili bir şekilde mücadele etmek için yeni tedavi yöntemlerine ve yenilikçi yaklaşımlara olan acil ihtiyacı vurgulamaktadır. Dolayısıyla, bu çalışmada HIF-1α'nın potansiyel inhibitörleri olan 7a ve 7b bileşiklerinin antikanser özelliklerinin araştırılması amaçlanmıştır. Çalışmada HTB-54 ve BEAS-2B hücre hatları kullanılmıştır. Yeni sentezlenen HIF inhibitörleri 7a ve 7b'nin normoksik ve hipoksik koşullar altında hücre canlılığı üzerindeki etkisini belirlemek için MTT hücre canlılığı deneyleri yapılmıştır. HIF1A'nın kantitatif ekspresyon seviyeleri, Real-Time PCR yöntemi ile belirlenmiştir. HTB-54 hücrelerinde 7a bileşiğinin yarı maksimum inhibitör konsantrasyonu (IC50) normoksik koşullar altında 10,37 µM iken hipoksik koşullar altında 10.63 µM olarak bulunmuştur. HTB-54 hücrelerindeki diğer bir HIF inhibitörü 7b'nin IC50 değeri normoksik koşullar altında 8,80 µM, hipoksik koşullar altında ise 9.54 µM olarak bulunmuştur. Hipoksik koşullarda, 7a ve 7b bileşikleri ile maruz bırakılan hücrelerde HIF1A ekspresyon düzeyinin kontrol grubuna göre daha düşük olduğu gösterilmiştir. Normoksi koşullarda ise HIF1A ekspresyon düzeyi 7a bileşiği ile maruz bırakılan hücrelerde kontrol grubuna göre 6,5 kat (p<0.0001), 7b'ye maruz kaldığında ise yaklaşık 9 kat (p<0.0001) arttığı bulunmuştur. Sonuç olarak hem 7a hem de 7b bileşikleri akciğer kanserine yönelik gelecekteki terapötik girişimler için büyük umut vaat etmektedir.

Anahtar Kelimeler

Akciğer Kanseri, HIF inhibitörleri, Hipoksi

Kaynakça

• 1. Jia XB, Zhang Q, Xu L, et al. Lotus leaf favonoids induce apoptosis of human lung cancer A549 cells through the ROS/p38 MAPK pathway. Biol Res. 2021;54:7.
• 2. Zhang C, Tang B, Hu J, et al. Neutrophils correlate with hypoxia microenvironment and promote progression of non-small-cell lung cancer. Bioengineered. 2021;12(1):8872–84.
• 3. Yu M, Qi B, Wu X, et al. Baicalein increases cisplatin sensitivity of A549 lung adenocarcinoma cells via PI3K/Akt/NF-kB pathway. Biomed Pharmacother. 2017;90:677-85.
• 4. He F, Wang G, Xu ZQ, et al. Nelfinavir restricts A549 cell growth by inhibiting STAT3 signaling. J Int Med Res. 2021;49(6):1–10.
• 5. Han M, Zhao Y, Tan C, et al. Cathepsin L upregulation-induced EMT phenotype is associated with the acquisition of cisplatin or paclitaxel resistance in A549 cells. Acta Pharmacol Sin. 2016;37:1606–22.
• 6. He R, Liu H. TRIM59 knockdown blocks cisplatin resistance in A549/DDP cells through regulating PTEN/AKT/HK2. Gene. 2020;747:144553.
• 7. Jing X, Yang F, Shao C, et al. Role of hypoxia in cancer therapy by regulating the tumor microenvironment. Mol Cancer. 2019;18:157.
• 8. Korbecki J, Siminska D, Dobrowolska MG, et al. Chronic and cycling hypoxia: Drivers of cancer chronic inflammation through HIF-1 and NF-κB activation: A review of the molecular mechanisms. Int J Mol Sci. 2021;22:10701.
• 9. Littlefower AB, Antony GR, Parambil ST, et al. Metabolic phenotype intricacies on altered glucose metabolism of breast cancer cells upon glut 1 inhibition and mimic hypoxia in vitro. Appl Biochem Biotechnol. 2023;195(10):5838-54.
• 10. Bozgeyik E, Kahraman-Taşdemir D, Arman K, et al. Novel thiosemicarbazone derivative 17B interferes with the cell cycle progression and induce apoptosis through modulating downstream signaling pathways. Gene Rep. 2020;18:100578.
• 11. Zhong X, Lin R, Li Z, et al. Effects of salidroside on cobalt chloride-induced hypoxia damage and mtor signaling repression in PC12 cells. Biol Pharm Bull. 2014;37(7):1199–206.
• 12. Gao XX, Liu CH, Hu ZL, et al. The biological effect of cobalt chloride mimetic-hypoxia on nucleus pulposus cells and the comparability with physical hypoxia in vitro. Front Biosci (Landmark Ed). 2021;26(10):799-812.
• 13. Ziello JE, Jovin IS, Huang Y. Hypoxia-Inducible Factor (HIF)-1 regulatory pathway and its potential for therapeutic intervention in malignancy and ischemia. Yale J Biol Med. 2007;80(2):51-60.
• 14. Jun CJ, Rathore A, Younas H, et al. Hypoxia-inducible factors and cancer. Curr Sleep Med Rep. 2017;3(1):1-10 15. Melillo Giovanni. Hypoxia-inducible factor 1 inhibitors. Methods Enzymol. 2007;435:385-402.
• 16. Ivan M, Haberberger T, Gervasi DC, et al. Biochemical purification and pharmacological inhibition of a mammalian prolyl hydroxylase acting on hypoxia-inducible factor. Proc Natl Acad Sci USA. 2002;99:13459–64.
• 17. Mun J, Jabbar AA, Devi NS, et al. Structure–activity relationship of 2,2-dimethyl-2H-chromene based arylsulfonamide analogs of 3,4-dimethoxy-N-[(2,2-dimethyl-2H-chromen-6-yl)methyl]-N-phenylbenzenesulfonamide, a novel small molecule hypoxia inducible factor-1 (HIF-1) pathway inhibitor and anti-cancer agent. Bioorg Med Chem. 2012;20(14):4590–7.
• 18. Rogers JL, Bayeh L, Scheuermann TH, et al. Development of inhibitors of the PAS-B domain of the HIF-2α transcription factor. J Med Chem. 2013;56(4):1739–47.
• 19. Taşdemir Kahraman D, Karaküçük İyidoan A, Saygideger Y, et al. Discovery of new chiral sulfonamides bearing benzoxadiazole as HIF inhibitors for non-small cell lung cancer therapy: design, microwave-assisted synthesis, binding affinity, in vitro antitumoral activities and in silico studies. New J Chem. 2022;46:2277-91.
• 20. Brahimi-Horn MC, Chiche J, Pouysségur J. Hypoxia and cancer. J Mol Med. 2007;85:1301-7.