Effects of Cyclosporine A and Adalimumab on the expression profiles histaminergic system-associated genes and microRNAs regulating these genes in HaCaT cells

Kasela, Tomasz; Dąbala, Maciej; Mistarz, Magdalena; Wieczorek, Weronika; Wierzbik-Strońska, Magdalena; Boroń, Kacper; Zawidlak-Węgrzyńska, Barbara; Grabarek, Beniamin Oskar

doi:10.1080/15384101.2022.2103342

Cited by 7 publications

(6 citation statements)

References 74 publications

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“…The control culture remained untreated with LPS and the drug. The selection of the adalimumab concentration was empirical, considering the average adalimumab concentration in the serum of psoriasis-treated patients, consistent with our previous research [ 31 , 32 ].…”

Section: Methodsmentioning

confidence: 63%

Expression profile of mRNAs and miRNAs related to mitogen-activated kinases in HaCaT cell culture treated with lipopolysaccharide a and adalimumab

Wójcik,

Plata-Babula,

Głowaczewska

et al. 2024

Cell Cycle

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Studies indicate that mitogen-activated protein kinases (MAPKs) exhibit activation and overexpression within psoriatic lesions. This study aimed to investigate alterations in the expression patterns of genes encoding MAPKs and microRNA (miRNA) molecules that potentially regulate their expression in human adult low-calcium high-temperature (HaCaT) keratinocytes when exposed to bacterial lipopolysaccharide A (LPS) and adalimumab. HaCaT cells underwent treatment with 1 µg/mL LPS for 8 hours, followed by treatment with 8 µg/mL adalimumab for 2, 8, or 24 hours. Untreated cells served as controls. The molecular analysis involved microarray, quantitative real-time polymerase chain reaction (RTqPCR), and enzyme-linked immunosorbent assay (ELISA) analyses. Changes in the expression profile of seven mRNAs: dual specificity phosphatase 1 ( DUSP1) , dual specificity phosphatase 3 ( DUSP3) , dual specificity phosphatase 4 ( DUSP4) , mitogen-activated protein kinase 9 ( MAPK9) , mitogen-activated protein kinase kinase kinase 2 ( MAP3K2) , mitogen-activated protein kinase kinase 2 ( MAP2K2), and MAP kinase-activated protein kinase 2 (MAPKAPK2 , also known as MK2) in cell culture exposed to LPS or LPS and the drug compared to the control. It was noted that miR-34a may potentially regulate the activity of DUSP1 , DUSP3 , and DUSP4 , while miR-1275 is implicated in regulating MAPK9 expression. Additionally, miR-382 and miR-3188 are potential regulators of DUSP4 levels, and miR-200-5p is involved in regulating MAPKAPK2 and MAP3K2 levels. Thus, the analysis showed that these mRNA molecules and the proteins and miRNAs they encode appear to be useful molecular markers for monitoring the efficacy of adalimumab therapy.

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Section: Methodsmentioning

confidence: 63%

Expression profile of mRNAs and miRNAs related to mitogen-activated kinases in HaCaT cell culture treated with lipopolysaccharide a and adalimumab

Wójcik,

Plata-Babula,

Głowaczewska

et al. 2024

Cell Cycle

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“…Culture untreated with LPS and the drug was the control culture. The concentration of CsA was selected empirically, taking into account the average concentration of CsA in the serum of patients treated with it for psoriasis, the same as in our previous work [ 21 , 22 ].…”

Section: Methodsmentioning

confidence: 99%

Gene expression profile of mitogen-activated kinases and microRNAs controlling their expression in HaCaT cell culture treated with lipopolysaccharide A and cyclosporine A

Wójcik,

Zmarzły,

Derkacz

et al. 2024

Cell Cycle

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Studies indicate that mitogen-activated protein kinases (MAPKs) are activated and overexpressed in psoriatic lesions. The aim of the study was to assess changes in the expression pattern of genes encoding MAPKs and microRNA (miRNA) molecules potentially regulating their expression in human adult low-calcium high-temperature (HaCaT) keratinocytes exposed to bacterial lipopolysaccharide A (LPS) and cyclosporine A (CsA). HaCaT cells were treated with 1 µg/mL LPS for 8 h, followed by treatment with 100 ng/mL cyclosporine A for 2, 8, or 24 h. Untreated cells served as controls. The molecular analysis consists of microarray, quantitative real-time polymerase chain reaction, and enzyme-linked immunosorbent assay analyses. The statistical analysis of the obtained results was performed using Transcriptome Analysis Console and STATISTICA 13.5 PL with the statistical significance threshold of p < 0.05. Changes in the expression profile of six mRNAs: dual-specificity phosphatase 1 ( DUSP1) , dual-specificity phosphatase 4 ( DUSP4) , mitogen-activated protein kinase kinase 2 ( MAP2K2) , mitogen-activated protein kinase kinase 7 ( MAP2K7) , mitogen-activated protein kinase kinase kinase 2 ( MAP3K2) and mitogen-activated protein kinase 9 ( MAPK9) in cell culture exposed to LPS or LPS and the drug compared to the control. We observed that under the LPS and cyclosporine treatment, the expression o/ miR-34a, miR-1275, miR-3188, and miR-382 changed significantly ( p < 0.05). We demonstrated a potential relationship between DUSP1 and miR-34a; DUSP4 and miR-34a, miR-382, and miR-3188; MAPK9 and miR-1275, MAP2K7 and mir-200-5p; MAP3K2 and mir-200-5p, which may be the subject of further research in the context of psoriasis.

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Section: Methodsmentioning

confidence: 99%

“…HaCaT cells were originally isolated from human adult skin and named based on their origin (human, adult, low-calcium, high-temperature, keratinocytes) [ 35 ]. HaCaT is considered a permanent non-malignant epithelial cell line, which maintains full epidermal differentiation capability [ 35 ]. Although it exhibits spontaneous phenotypic transformation and despite the unlimited growth potential, HaCaT cells, similar to normal keratinocytes, it reforms an orderly structured and differentiated epidermal tissue when transplanted onto nude mice, remaining non-tumorigenic [ 32 ].…”

Section: Methodsmentioning

confidence: 99%

“…HaCaT is a human adult, low-calcium, high-temperature, keratinocyte cell line, which represents a cellular model commonly used to study normal keratinocytes [ 32 , 33 , 34 ]. The cells exhibit, similar to human keratinocytes, the following features: physiological keratinocyte morphology, epidermal differentiation capability, non-tumorigenic, and can undergo ultraviolet light-induced apoptosis [ 35 ]. Keratinocytes, indeed, can impact melanoma progression as they belong to tumor microenvironment by closely interacting with melanocytes in skin melanoma.…”

Section: Introductionmentioning

confidence: 99%

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Human Melanoma Cells Differentially Express RNASEL/RNase-L and miR-146a-5p under Sex Hormonal Stimulation

Orlandi

Tomi

Campagnari

et al. 2022

CIMB

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Polymorphisms in the ribonuclease L (RNASEL) coding gene and hsa-miR-146a-5p (miR-146a) have been associated with melanoma in a sex-specific manner. We hypothesized that RNASEL and miR-146a expression could be influenced by sex hormones playing a role in the female advantages observed in melanoma incidence and survival. Thus, we explored the effects of testosterone and 17β-estradiol on RNASEL and miR-146a expression in LM-20 and A375 melanoma cell lines. Direct targeting of miR-146a to the 3’ untranslated region (3′UTR) of RNASEL was examined using a luciferase reporter system. Our results indicate that RNASEL is a direct target of miR-146a in both melanoma cell lines. Trough qPCR and western blot analyses, we explored the effect of miR-146a mimic transfection in the presence of each hormone either on RNASEL mRNA level or on protein expression of RNase-L, the enzyme codified by RNASEL gene. In the presence of testosterone or 17β-estradiol, miR-146a overexpression did not influence RNASEL transcript level in LM-20 cell line, but it slightly induced RNASEL mRNA level in A375 cells. Remarkably, miR-146a overexpression was able to repress the protein level of RNase-L in both LM-20 and A375 cells in the presence of each hormone, as well as to elicit high expression levels of the activated form of the extracellular signal-regulated kinases (ERK)1/2, hence confirming the pro-tumorigenic role of miR-146a overexpression in melanoma. Thereafter, we assessed if the administration of each hormone could affect the endogenous expression of RNASEL and miR-146a genes in LM-20 and A375 cell lines. Testosterone exerted no significant effect on RNASEL gene expression in both cell lines, while 17β-estradiol enhanced RNASEL transcript level at least in LM-20 melanoma cells. Conversely, miR-146a transcript augmented only in the presence of testosterone in either melanoma cell line. Importantly, each hormone acted quite the opposite regarding the RNase-L protein expression, i.e., testosterone significantly decreased RNase-L expression, whereas 17β-estradiol increased it. Overall, the data show that, in melanoma cells treated with 17β-estradiol, RNase-L expression increased likely by transcriptional induction of its gene. Testosterone, instead, decreased RNase-L expression in melanoma cell lines with a post-transcriptional mechanism in which miR-146a could play a role. In conclusion, the pro-tumor activity of androgen hormone in melanoma cells could be exacerbated by both miR-146a increase and RNase-L downregulation. These events may contribute to the worse outcome in male melanoma patients.

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Effects of Cyclosporine A and Adalimumab on the expression profiles histaminergic system-associated genes and microRNAs regulating these genes in HaCaT cells

Cited by 7 publications

References 74 publications

Expression profile of mRNAs and miRNAs related to mitogen-activated kinases in HaCaT cell culture treated with lipopolysaccharide a and adalimumab

Expression profile of mRNAs and miRNAs related to mitogen-activated kinases in HaCaT cell culture treated with lipopolysaccharide a and adalimumab

Gene expression profile of mitogen-activated kinases and microRNAs controlling their expression in HaCaT cell culture treated with lipopolysaccharide A and cyclosporine A

Human Melanoma Cells Differentially Express RNASEL/RNase-L and miR-146a-5p under Sex Hormonal Stimulation

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