Transcriptome Profiling of Cardiac Glycoside Treatment Reveals EGR1 and Downstream Proteins of MAPK/ERK Signaling Pathway in Human Breast Cancer Cells
Cardiac glycosides (CGs) constitute a group of steroid-like compounds renowned for their effectiveness in treating cardiovascular ailments. In recent times, there has been growing recognition of their potential use as drug leads in cancer treatment. In our prior research, we identified three highly...
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| Format: | Article |
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MDPI AG
2023-11-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/21/15922 |
| _version_ | 1797631768750194688 |
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| author | Honey Pavithran Ranjith Kumavath Preetam Ghosh |
| author_facet | Honey Pavithran Ranjith Kumavath Preetam Ghosh |
| author_sort | Honey Pavithran |
| collection | DOAJ |
| description | Cardiac glycosides (CGs) constitute a group of steroid-like compounds renowned for their effectiveness in treating cardiovascular ailments. In recent times, there has been growing recognition of their potential use as drug leads in cancer treatment. In our prior research, we identified three highly promising CG compounds, namely lanatoside C (LC), peruvoside (PS), and strophanthidin (STR), which exhibited significant antitumor effects in lung, liver, and breast cancer cell lines. In this study, we investigated the therapeutic response of these CGs, with a particular focus on the MCF-7 breast cancer cell line. We conducted transcriptomic profiling and further validated the gene and protein expression changes induced by treatment through qRT-PCR, immunoblotting, and immunocytochemical analysis. Additionally, we demonstrated the interactions between the ligands and target proteins using the molecular docking approach. The transcriptome analysis revealed a cluster of genes with potential therapeutic targets involved in cytotoxicity, immunomodulation, and tumor-suppressor pathways. Subsequently, we focused on cross-validating the ten most significantly expressed genes, <i>EGR1</i>, <i>MAPK1</i>, <i>p53</i>, <i>CCNK</i>, <i>CASP9</i>, <i>BCL2L1</i>, <i>CDK7</i>, <i>CDK2</i>, <i>CDK2AP1</i>, and <i>CDKN1A</i>, through qRT-PCR, and their by confirming the consistent expression pattern with RNA-Seq data. Notably, among the most variable genes, we identified EGR1, the downstream effector of the MAPK signaling pathway, which performs the regulatory function in cell proliferation, tumor invasion, and immune regulation. Furthermore, we substantiated the influence of CG compounds on translational processes, resulting in an alteration in protein expression upon treatment. An additional analysis of ligand–protein interactions provided further evidence of the robust binding affinity between LC, PS, and STR and their respective protein targets. These findings underscore the intense anticancer activity of the investigated CGs, shedding light on potential target genes and elucidating the probable mechanism of action of CGs in breast cancer. |
| first_indexed | 2024-03-11T11:27:02Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T11:27:02Z |
| publishDate | 2023-11-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-122581f7fd504c3fa8f710b582c61f192023-11-10T15:05:48ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-11-0124211592210.3390/ijms242115922Transcriptome Profiling of Cardiac Glycoside Treatment Reveals EGR1 and Downstream Proteins of MAPK/ERK Signaling Pathway in Human Breast Cancer CellsHoney Pavithran0Ranjith Kumavath1Preetam Ghosh2Department of Genomic Science, School of Biological Sciences, Central University of Kerala, Kasaragod 671320, IndiaDepartment of Genomic Science, School of Biological Sciences, Central University of Kerala, Kasaragod 671320, IndiaDepartment of Computer Science, Virginia Commonwealth University, Richmond, VA 23284, USACardiac glycosides (CGs) constitute a group of steroid-like compounds renowned for their effectiveness in treating cardiovascular ailments. In recent times, there has been growing recognition of their potential use as drug leads in cancer treatment. In our prior research, we identified three highly promising CG compounds, namely lanatoside C (LC), peruvoside (PS), and strophanthidin (STR), which exhibited significant antitumor effects in lung, liver, and breast cancer cell lines. In this study, we investigated the therapeutic response of these CGs, with a particular focus on the MCF-7 breast cancer cell line. We conducted transcriptomic profiling and further validated the gene and protein expression changes induced by treatment through qRT-PCR, immunoblotting, and immunocytochemical analysis. Additionally, we demonstrated the interactions between the ligands and target proteins using the molecular docking approach. The transcriptome analysis revealed a cluster of genes with potential therapeutic targets involved in cytotoxicity, immunomodulation, and tumor-suppressor pathways. Subsequently, we focused on cross-validating the ten most significantly expressed genes, <i>EGR1</i>, <i>MAPK1</i>, <i>p53</i>, <i>CCNK</i>, <i>CASP9</i>, <i>BCL2L1</i>, <i>CDK7</i>, <i>CDK2</i>, <i>CDK2AP1</i>, and <i>CDKN1A</i>, through qRT-PCR, and their by confirming the consistent expression pattern with RNA-Seq data. Notably, among the most variable genes, we identified EGR1, the downstream effector of the MAPK signaling pathway, which performs the regulatory function in cell proliferation, tumor invasion, and immune regulation. Furthermore, we substantiated the influence of CG compounds on translational processes, resulting in an alteration in protein expression upon treatment. An additional analysis of ligand–protein interactions provided further evidence of the robust binding affinity between LC, PS, and STR and their respective protein targets. These findings underscore the intense anticancer activity of the investigated CGs, shedding light on potential target genes and elucidating the probable mechanism of action of CGs in breast cancer.https://www.mdpi.com/1422-0067/24/21/15922breast cancerRNA-Seq analysislanatoside Cperuvosidestrophanthidinanticancer activity |
| spellingShingle | Honey Pavithran Ranjith Kumavath Preetam Ghosh Transcriptome Profiling of Cardiac Glycoside Treatment Reveals EGR1 and Downstream Proteins of MAPK/ERK Signaling Pathway in Human Breast Cancer Cells International Journal of Molecular Sciences breast cancer RNA-Seq analysis lanatoside C peruvoside strophanthidin anticancer activity |
| title | Transcriptome Profiling of Cardiac Glycoside Treatment Reveals EGR1 and Downstream Proteins of MAPK/ERK Signaling Pathway in Human Breast Cancer Cells |
| title_full | Transcriptome Profiling of Cardiac Glycoside Treatment Reveals EGR1 and Downstream Proteins of MAPK/ERK Signaling Pathway in Human Breast Cancer Cells |
| title_fullStr | Transcriptome Profiling of Cardiac Glycoside Treatment Reveals EGR1 and Downstream Proteins of MAPK/ERK Signaling Pathway in Human Breast Cancer Cells |
| title_full_unstemmed | Transcriptome Profiling of Cardiac Glycoside Treatment Reveals EGR1 and Downstream Proteins of MAPK/ERK Signaling Pathway in Human Breast Cancer Cells |
| title_short | Transcriptome Profiling of Cardiac Glycoside Treatment Reveals EGR1 and Downstream Proteins of MAPK/ERK Signaling Pathway in Human Breast Cancer Cells |
| title_sort | transcriptome profiling of cardiac glycoside treatment reveals egr1 and downstream proteins of mapk erk signaling pathway in human breast cancer cells |
| topic | breast cancer RNA-Seq analysis lanatoside C peruvoside strophanthidin anticancer activity |
| url | https://www.mdpi.com/1422-0067/24/21/15922 |
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