P53/MDM2 Co-Expression in Laryngeal Squamous Cell Carcinoma Based on Digital Image Analysis

Chrysovergis, Aristeidis; Papanikolaou, Vasileios; Tsiambas, Evangelos; Stavraka, Chara; Ragos, Vasileios; Peschos, Dimitrios; Psyrri, Amanda; Mastronikolis, Nicholas; Kyrodimos, Efthymios

doi:10.21873/anticanres.13572

Cited by 10 publications

(7 citation statements)

References 28 publications

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“…Sendtner et al [14] found that upregulation of LIF significantly increased nutritional and physiological related effects on motoneurons by cotransfection of CNTF and LIF genes. Chrysovergis et al [23] also proved that double-gene integration promoted the expression of target genes in laryngeal squamous cell carcinoma. The reason may be related to the gene regulatory network in the organism and the complex and multilevel regulation of gene expression.…”

Section: Discussionmentioning

confidence: 97%

Double-Gene Copromoting Expression Analysis in tPA/GH Transgenic Goat Mammary Epithelial Cells and Thrombolytic Activity of tPA In Vitro

Song

et al. 2022

BioMed Research International

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Human tissue-plasminogen activator (tPA) is a thrombolytic drug widely used in the treatment of stroke, pulmonary thrombosis, acute myocardial infarction, and other thrombotic diseases. The double genes cointegrated into the organisms and cells can produce a synergistic effect, which will improve the expression level of the target gene. However, the study of the integration of the GH and tPA genes to improve the expression level of tPA has not yet been reported. In order to elucidate this, we generated monoclonal goat mammary epithelial cell lines with tPA/GH double-gene integration and analyzed the tPA expression level in single- and double-gene integrated cells. We selected the mammary gland-specific expressing vectors BLC14/tPA and BLC14/GH with the β-lactoglobulin gene as a regulatory sequence in our previous research. The tPA and GH genes were electronically cotransfected into goat mammary epithelial cells. Resistant cell lines were screened by G418, and transgenic monoclonal cell lines were confirmed by PCR. The tPA expression was induced by prolactin and detected in the cell induction solution after 48 h by ELISA and Western blotting. We detected the tPA biological activity in vitro by fibrin agarose plate assay (FAPA). The results showed that a total of 207 resistant monoclonal cells were obtained, including 126 cell lines with tPA monogenic integration and 51 cell lines with tPA/GH double-gene integration. The rate of double-gene integration was 24.6% (51/207). A total of 48 cells expressed tPA, of which 25.3% (19/75) cells expressed single gene, and 56.9% (29/51) cells expressed double genes. The concentration of tPA in single-gene-expressing cells was 8.0-64.0 μg/mL, and the tPA level in double-gene-expressing cells was significantly higher (200-7200 μg/mL). In addition, the tPA had a relatively strong in vitro thrombolytic activity determined by FAPA. The results showed that goat mammary epithelial cell lines with tPA/GH gene integration were successfully established by electrotransfection, and the expression level of tPA in double-gene integrated cell lines was significantly increased. This study provided a new way for the preparation of a transgenic goat and other animal with high tPA expression by somatic cell nuclear transfer. The findings also laid a foundation for efficient production of pharmaceutical proteins in transgenic animal mammary gland bioreactors in the future.

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Section: Discussionmentioning

confidence: 97%

Double-Gene Copromoting Expression Analysis in tPA/GH Transgenic Goat Mammary Epithelial Cells and Thrombolytic Activity of tPA In Vitro

Song

et al. 2022

BioMed Research International

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“…For the past few years, many biomarkers have been applied for the diagnosis and treatment of LSCC (Cossu et al, 2019). Various anti-LSCC mechanisms have also been identified (Chrysovergis et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

“…In addition, a recent study recorded that cytochrome P450 inhibited the activity of the metabolic enzymes CYP2C9 Ã 2 and CYP2C9 Ã 3, which could directly control tumorigenesis by reducing epoxyeicosatrienoic acid production (Hunter et al, 2015;Katiyar et al, 2017). The PI3K-Akt signalling pathway has a vital function in LSCC by suppressing cell death (Chrysovergis et al, 2019;Yang et al, 2019). P53, a tumour suppressor factor, initiates DNA repair, cell cycle arrest and apoptosis and reacts to various kinds of cancer therapies (Cui, Qu & Liu, 2019;Ragos et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

Bioinformatics analysis of laryngeal squamous cell carcinoma: seeking key candidate genes and pathways

Ma¹,

Hu²,

Dai³

et al. 2021

PeerJ

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Background Laryngeal squamous cell carcinoma (LSCC) is the second most aggressive head and neck squamous cell carcinoma. Although much work has been done to optimize its treatment, patients with LSCC still have poor prognosis. Therefore, figuring out differentially expressed genes (DEGs) contained in the progression of LSCC and employing them as potential therapeutic targets or biomarkers for LSCC is extremely meaningful. Methods Overlapping DEGs were screened from two standalone Gene Expression Omnibus datasets, and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed. By applying STRING and Cytoscape, a protein–protein network was built, and module analysis was carried out. The hub genes were selected by maximal clique centrality with the CytoHubba plugin of Cytoscape. UALCAN and GEPIA data were examined to validate the gene expression findings. Moreover, the connection of the hub genes with LSCC patient overall survival was studied employing The Cancer Genome Atlas. Then, western blot, qRT-PCR, CCK-8, wound healing and transwell assays were bring to use for further verify the key genes. Results A total of 235 DEGs were recorded, including 83 upregulated and 152 downregulated genes. A total of nine hub genes that displayed a high degree of connectivity were selected. UALCAN and GEPIA databases verified that these genes were highly expressed in LSCC tissues. High expression of the SPP1, SERPINE1 and Matrix metalloproteinases 1 (MMP1) genes was connected to worse prognosis in patients with LSCC, according to the GEPIA online tool. Western blot and qRT-PCR testify SPP1, SERPINE1 and MMP1 were upregulated in LSCC cells. Inhibition of SPP1, SERPINE1 and MMP1 suppressed cell proliferation, invasion and migration. Conclusion The work here identified effective and reliable diagnostic and prognostic molecular biomarkers by unified bioinformatics analysis and experimental verification, indicating novel and necessary therapeutic targets for LSCC.

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“…Moreover, p53 expression, together with Mouse double minute 2 homolog expression (MDM2), was positively correlated with advanced stage of LSCC by Chrysovergis et al, as demonstrated by immunostaining of p53 and MDM2 in the 50 LSCC patients included in their study. The results showed p53 overexpression in 32% LSCC patients and a stronger MDM2 expression in 44% cases [ 33 ]. TP53 gene mutations have been also assumed to affect the response to radiotherapy and drive both cell differentiation and neck LNM, although no correlation with LSCC clinical stages was observed [ 34 , 35 ].…”

Section: Molecular Markers For Clinical Management Of Lsccmentioning

confidence: 99%

Overview on Molecular Biomarkers for Laryngeal Cancer: Looking for New Answers to an Old Problem

Falco

Tammaro

Takeuchi

et al. 2022

Cancers

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Laryngeal squamous cell cancer (LSCC) accounts for almost 25–30% of all head and neck squamous cell cancers and is clustered according to the affected districts, as this determines distinct tendency to recur and metastasize. A major role for numerous genetic alterations in driving the onset and progression of this neoplasm is emerging. However, major efforts are still required for the identification of molecular markers useful for both early diagnosis and prognostic definition of LSCC that is still characterized by significant morbidity and mortality. Non-coding RNAs appear the most promising as they circulate in all the biological fluids allowing liquid biopsy determination, as well as due to their quick and characteristic modulation useful for non-invasive detection and monitoring of cancer. Other critical aspects are related to recent progress in circulating tumor cells and DNA detection, in metastatic status and chemo-refractoriness prediction, and in the functional interaction of LSCC with chronic inflammation and innate immunity. We review all these aspects taking into account the progress of the technologies in the field of next generation sequencing.

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P53/MDM2 Co-Expression in Laryngeal Squamous Cell Carcinoma Based on Digital Image Analysis

Cited by 10 publications

References 28 publications

Double-Gene Copromoting Expression Analysis in tPA/GH Transgenic Goat Mammary Epithelial Cells and Thrombolytic Activity of tPA In Vitro

Double-Gene Copromoting Expression Analysis in tPA/GH Transgenic Goat Mammary Epithelial Cells and Thrombolytic Activity of tPA In Vitro

Bioinformatics analysis of laryngeal squamous cell carcinoma: seeking key candidate genes and pathways

Overview on Molecular Biomarkers for Laryngeal Cancer: Looking for New Answers to an Old Problem

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