Expression Levels of ACE2 and TMPRSS2 in Different Cell Lines

Year 2023, , 260 - 268, 30.06.2023

Merve Gülsen Bal Albayrak , Sevinc Yanar , Murat Kasap , Gürler Akpınar

https://doi.org/10.53446/actamednicomedia.1253701

Cited By: 1

Abstract

Objective: ACE2 and TMPRSS2 proteins have received increased attention gained emphasis together with the pandemic COVID-19. These proteins have roles in respiratory and hypertension disorders as well as cardiovascular and renal diseases. The objective of this work was to examine the mRNA and protein levels of ACE2 and TMPRSS2 in cell lines derived from various tissue origins.
Methods: After the growth of 14 different cell lines, protein and mRNA were isolated from the cell pellets. The amounts of mRNAs and proteins were then determined and quantified using RT-PCR and ELISA.
Results: Findings showed that VERO, HEK293T, and VERO E6 cell lines significantly differed from others in the mRNA levels of both the ACE2 and TMPRSS2 genes. In protein levels obtained using ELISA, PNT1A cell line had the highest level of ACE2 protein expression, while for TMPRSS2, A549 had the highest level of protein expression.
Conclusion: It was showed in this study how the expressions of ACE2 and TMPRSS2 depend on the cell type. This may be an explanation for why virulence entrance differs in different types of tissues. It is thought that HEK293T cells with high levels of both genes may be a suitable option for studies at the RNA level by using these two genes. MCF7 may be a good option for studies at the protein level. Given the high levels of mRNA expression of both genes, it may be inferred that cells derived from the kidney were among those that were most susceptible to virus entry.

Keywords

ACE2, TMPRSS2, COVID19

ACE2 ve TMPRSS2 Genlerinin Farklı Hücre Hatlarındaki İfade Düzeyleri

Abstract

Amaç: ACE2 ve TMPRSS2 proteinleri, COVID-19 pandemisi ile birlikte önem kazanmıştır. Bu proteinlerin solunum ve hipertansiyon bozukluklarının yanı sıra kardiyovasküler ve renal hastalıklarda rolleri vardır. Bu çalışmanın amacı, çeşitli dokulardan üretilen hücre hatlarında ACE2 ve TMPRSS2'nin mRNA ve protein seviyelerini belirlemektir.
Yöntem: 14 hücre hattı kültürde çoğaltıldıktan sonra, hücre pelletlerinden protein ve mRNA izole edildi. Ardından mRNA seviyeleri RT-PCR ve protein seviyeleri ise ELISA metotları kullanılarak ölçüldü.
Bulgular: Çalışmanın sonuçları, ACE2 ve TMPRSS2 genlerinin mRNA seviyelerinin VERO, HEK293T ve VERO E6 hücre hatlarında, diğer hücre hatlarına kıyasla, önemli ölçüde yüksek olduğunu göstermiştir. ELISA kullanılarak elde edilen protein seviyelerinde, PNT1A hücre hattı en yüksek ACE2 protein ekspresyonuna sahipken, TMPRSS2 için A549 en yüksek protein ekspresyonuna sahip olarak bulunmuştur.
Sonuç: Bu çalışmada ACE2 ve TMPRSS2 gen ifadelerinin hücre tipine göre farklılaşabileceği gösterilmiştir. Bu sonuç, farklı doku tiplerinde virüs girişinin neden farklı olduğunun bir açıklaması olabilir. Her iki genin de yüksek düzeyde bulunduğu HEK293T hücrelerinin, bu iki gen kullanılarak RNA düzeyinde yapılacak çalışmalar için uygun bir seçenek olabileceği düşünülmektedir. MCF7 hücre hattı ise protein seviyesindeki çalışmalar için iyi bir seçenek olabilir. Her iki genin yüksek mRNA ekspresyon seviyeleri göz önüne alındığında, böbrekten üretilen hücre hatlarının virüs girişine en duyarlı olanlar arasında olduğu sonucuna varılabilir.

Keywords

ACE2, TMPRSS2, COVID19

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