Formaldehyde induces toxic effects and regulates the expression of damage response genes in BM-MSCs

Abstract

In this study, we assessed the toxic effects of formaldehyde (FA) on mouse bone marrow mesenchymal stem cells (BMMSCs). Cytotoxicity was measured by using MTT assay. DNA strand breakage was detected by standard alkaline comet assay and comet assay modified with proteinase K (PK). DNA -protein crosslinks (DPCs) were detected by KCl-SDS precipitation assay. We found that FA at a concentration from 75 to 200 mM inhibited cell survival and induced DPCs over 125 mM. The PK-modified comet assay showed that FA-induced DNA strand breakage was increased in a dose-dependent manner from 75 to 200 mM. On the other hand, standard alkaline comet assay showed that DNA strand breakage was decreased with FA concentration over 125 mM. We confirmed by using Pearson correlation that there was a negative linear correlation between DPCs and survival rate (r 5 20.987, P < 0.01) and positive linear relationships between DPCs and (i) sister chromatid exchange and (ii) micronucleus (r 5 0.995, P < 0.01; r 5 0.968, P < 0.01). DNA damage RT 2 profiler polymerase chain reaction array was used to investigate the changes in the expression of damage response genes. Xpa and Xpc of the nucleotide excision repair pathway and Brca2, Rad51, and Xrcc2 of the homologous recombination pathway were all up-regulated in both 75 and 125 mM FA. However, the same genes were down-regulated with 175 mM FA. The expressions of Chek1 and Hus1, which are involved in cell cycle regulation, were altered in the same manner with 75, 125, and 175 mM FA. These results indicated that Xpa, Xpc, Brca2, Rad51, Xrcc2, Chek1, and Hus1 were essential for the BM-MSCs to counteract the effects of FA.

Astragalus polysaccharide protects formaldehyde-induced toxicity by promoting NER pathway in bone marrow mesenchymal stem cells

“…Our study shows that the mRNA and protein expression levels of XPC and ERCC1 in FA-treated BMSCs were significantly higher than those in the control cells, indicating that XPC and ERCC1 may be involved in the repair of human BMSCs DNA damage caused by FA. This is accordance with previous studies [7,39]. The mRNA and protein expression levels of XPA, XPC, and ERCC1 in cells incubated with FA plus APS groups at different concentrations were significantly higher than those in the FA-treated cells.…”

“…Our previous studies have confirmed that formaldehyde (FA) could inhibit the proliferative activity and enhance DNA-protein crosslinks (DPCs) formation in mice bone marrow mesenchymal stem cells (BMSCs) [7]. Therefore, the main purpose of this study was to investigate the protective effect of Astragalus polysaccharide (APS) against cytotoxicity and genotoxicity in human BMSCs exposed to FA and its potential mechanisms.…”

“…FA exhibits toxic effects in various types of mammalian cells, including cultured human myeloid progenitor cells and bone marrow blood progenitor cells in vivo [5,6]. In our previous study, it has been confirmed that FA induced the excessive DNA-protein crosslinks (DPCs) in murine BMSCs [7]. The formation of DPCs is related to oxidative damage caused by FA-induced excessive hydroxyl free radicals [8][9][10].…”

SERPINB2 is a novel indicator of stem cell toxicity

“…Indeed, embryonic or normal tissue-derived stem cells and the patient’s own adult stem cells can be obtained and differentiated into various target tissues, including cardiomyocytes, hepatocytes, or neurons, for in vitro potential toxicity screening 21 . Especially, MSCs have been used for assessing the toxicity of various chemicals such as anesthetics 22 , 23 , chemotherapeutic drugs 24 , 25 , endocrine disruptors 26 , nanoparticles 27 , 28 , and other industrial chemicals including cosmetic ingredients or detergent 29 , 30 . Nevertheless, the comparison among the toxicological studies using MSCs is challenging, since laboratories around the world lack internationally standardized methods for isolation and in vitro expansion of MSCs, resulting in significant functional heterogeneity and inconsistent testing results.…”

Bone Marrow Stem Cells and Genotoxicity Studies in These Cells: Review

“…73,74 Fare mezenkimal kök hücrelerinde formaldehitin çeşitli dozlarda uygulamasının yapıldığı in vitro bir çalışmada, 125 m ve üzeri dozlarda DNAprotein çapraz bağlanmaları, zincir kırıkları ve DNA hasar yanıtında rol alan Brca2, Rad51, Xrcc2, Xpa, Xpc, Chek1 ve Hus1 düzeylerinde artma gö-rülmüştür. 75 Tümör oluşumunun kök hücrelerdeki spontan mutasyonlardan kaynaklandığı hipotezinin araştırıldığı bir çalışmada fare kemik iliği mezenkimal kök hücreleri, in vitro kültür ile çok sayıda pasajı yapılarak, malign hücrelere dönüşüm aşamasında farelere enjekte edilmiş ve fibrosarkom oluşumu gözlenmiştir. Mezenkimal hücrelerin malign hücrelere dönüşümünün kromozom anormallikleri, artan telomeraz aktivitesi ve c-myc ekpresyonuna bağlı olarak gerçekleştiği sonucuna varılmıştır.…”