Immunohistochemical analysis of pro- and active-caspase 3 in laryngeal squamous cell carcinoma

Cancer bioinformatics has been used to screen possible key cancer genes and pathways. Here, through bioinformatics analysis, we found that high expression of diaphanous related formin 1 (DIAPH1) was associated with poor overall survival in head and neck squamous cell carcinoma and laryngeal squamous cell carcinoma (LSCC). The effect of DIAPH1 in LSCC has not been previously investigated. Therefore, we evaluated the expression, function, and molecular mechanisms of DIAPH1 in LSCC. Immunohistochemistry and western blot analysis confirmed the significant upregulation of DIAPH1 in LSCC. We used DIAPH1 RNA interference to construct two DIAPH1-knockdown LSCC cell lines, AMC-HN-8 and FD-LSC-1, and validated the knockdown efficiency. Flow cytometry data showed that DIAPH1 inhibited apoptosis. Further, western blot analysis revealed that DIAPH1 knockdown increased the protein levels of ATR, p-p53, Bax, and cleaved caspase-3, -8, and -9. Thus, DIAPH1 is upregulated in LSCC and may act as an oncogene by inhibiting apoptosis through the ATR/p53/caspase-3 pathway in LSCC cells.

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Silencing of miR-17-5p suppresses cell proliferation and promotes cell apoptosis by directly targeting PIK3R1 in laryngeal squamous cell carcinoma

Cancer Cell International ◽

10.1186/s12935-020-1096-3 ◽

2020 ◽

Vol 20 (1) ◽

Cited By ~ 5

Author(s):

Jian-Xing Wang ◽

Xin-Ju Jia ◽

Yan Liu ◽

Jin-Hui Dong ◽

Xiu-Min Ren ◽

...

Keyword(s):

Cell Proliferation ◽

Squamous Cell Carcinoma ◽

Tumor Progression ◽

Cell Carcinoma ◽

Cell Lines ◽

Squamous Cell ◽

Cell Apoptosis ◽

Caspase 3 ◽

Laryngeal Squamous Cell Carcinoma

Abstract Background Increasing evidence has suggested that microRNAs (miRNAs) act as key post-transcriptional regulators in tumor progression. Previous studies have confirmed that miR-17-5p functions as an oncogene in multiple cancers and contributes to tumor progression. However, the role and biological functions of miR-17-5p in the development of laryngeal squamous cell carcinoma (LSCC) still remain unknown. Methods qRT-PCR was used to detect miRNA and mRNA expression levels in LSCC tissues and cell lines. CCK-8 assay was used to measure cell viability and flow cytometry was performed to evaluate cell apoptosis. Western blot analysis was used to detect the protein levels of BAX, BCL-2, cleaved Caspase-3, PIK3R1 and AKT. Luciferase reporter assay was used to detect the effect of miR-17-5p on PIK3R1 expression. Xenograft animal model was used to test the effect of miR-17-5p on LSCC cell in vivo. Results In the present study, we found that miR-17-5p expression level was upregulated in LSCC tissues and cell lines. Depletion of miR-17-5p in LSCC cells significantly reduced cell proliferation and promoted cell apoptosis in vitro and in vivo. Mechanically, knockdown of miR-17-5p in LSCC cells inhibited BCL-2 expression while enhanced BAX and cleaved Caspase-3 protein expression. Moreover, depletion of miR-17-5p in LSCC cells suppressed AKT phosphorylation but did not influence PTEN expression. Importantly, miR-17-5p positively regulated PIK3R1 expression by directly binding to its 3′-untranslated region (UTR). Additionally, PIK3R1, which expression was downregulated in LSCC tissues and cell lines, was involved in LSCC cell survival by modulating the activation of AKT signal pathway. Dysregulation of miR-17-5p/PIK3R1 axis was participated in LSCC cell proliferation and apoptosis by inhibiting the activation of the PI3K/AKT signaling pathway. Conclusions In conclusion, our study indicates that the miR-17-5p/PIK3R1 axis plays an essential role in the development of LSCC and provides a potential therapeutic target for LSCC treatment.

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The primary growth of laryngeal squamous cell carcinoma cells in vitro is effectively supported by paired cancer-associated fibroblasts alone

Tumor Biology ◽

10.1177/1010428317705512 ◽

2017 ◽

Vol 39 (5) ◽

pp. 101042831770551 ◽

Cited By ~ 5

Author(s):

Mei Wang ◽

Chunping Wu ◽

Yu Guo ◽

Xiaojuan Cao ◽

Wenwei Zheng ◽

...

Keyword(s):

Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Laryngeal Cancer ◽

Squamous Cell ◽

Caspase 3 ◽

Laryngeal Squamous Cell Carcinoma ◽

Carcinoma Cells ◽

Cancer Associated Fibroblasts ◽

Protein Levels

Most primarily cultured laryngeal squamous cell carcinoma cells are difficult to propagate in vitro and have a low survival rate. However, in our previous work to establish a laryngeal squamous cell carcinoma cell line, we found that laryngeal cancer-associated fibroblasts appeared to strongly inhibit the apoptosis of primarily cultured laryngeal squamous cell carcinoma cells in vitro. In this study, we investigated whether paired laryngeal cancer-associated fibroblasts alone can effectively support the growth of primarily cultured laryngeal squamous cell carcinoma cells in vitro. In all, 29 laryngeal squamous cell carcinoma specimens were collected and primarily cultured. The laryngeal squamous cell carcinoma cells were separated from cancer-associated fibroblasts by differential trypsinization and continuously subcultured. Morphological changes of the cultured laryngeal squamous cell carcinoma cells were observed. Immunocytofluorescence was used to authenticate the identity of the cancer-associated fibroblasts and laryngeal squamous cell carcinoma cells. Flow cytometry was used to quantify the proportion of apoptotic cells. Western blot was used to detect the protein levels of caspase-3. Enzyme-linked immunosorbent assay was used to detect the levels of chemokine (C-X-C motif) ligand 12, chemokine (C-X-C motif) ligand 7, hepatocyte growth factor, and fibroblast growth factor 1 in the supernatants of the laryngeal squamous cell carcinoma and control cells. AMD3100 (a chemokine (C-X-C motif) receptor 4 antagonist) and an anti–chemokine (C-X-C motif) ligand 7 antibody were used to block the tumor-supporting capacity of cancer-associated fibroblasts. Significant apoptotic changes were detected in the morphology of laryngeal squamous cell carcinoma cells detached from cancer-associated fibroblasts. The percentage of apoptotic laryngeal squamous cell carcinoma cells and the protein levels of caspase-3 increased gradually in subsequent subcultures. In contrast, no significant differences in the proliferation capacity of laryngeal squamous cell carcinoma cells cocultured with cancer-associated fibroblasts were detected during subculturing. High level of chemokine (C-X-C motif) ligand 12 was detected in the culture supernatant of cancer-associated fibroblasts. The tumor-supporting effect of cancer-associated fibroblasts was significantly inhibited by AMD3100. Our findings demonstrate that the paired laryngeal cancer-associated fibroblasts alone are sufficient to support the primary growth of laryngeal squamous cell carcinoma cells in vitro and that the chemokine (C-X-C motif) ligand 12/chemokine (C-X-C motif) receptor 4 axis is one of the major contributors.

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Immunohistochemical Analysis and Prognostic Value of Cathepsin D Determination in Laryngeal Squamous Cell Carcinoma

Journal of Chemical Information and Computer Sciences ◽

10.1021/ci990075q ◽

2000 ◽

Vol 40 (3) ◽

pp. 545-549 ◽

Cited By ~ 3

Author(s):

Sven Seiwerth ◽

Nikola Štambuk ◽

Paško Konjevoda ◽

Nikola Mašić ◽

Ankica Vasilj ◽

...

Keyword(s):

Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Squamous Cell ◽

Cathepsin D ◽

Prognostic Value ◽

Laryngeal Squamous Cell Carcinoma ◽

Immunohistochemical Analysis

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The Role ofH. pyloriin the Development of Laryngeal Squamous Cell Carcinoma

Disease Markers ◽

10.1155/2013/950701 ◽

2013 ◽

Vol 35 ◽

pp. 447-449 ◽

Cited By ~ 6

Author(s):

Raşan Genç ◽

Sedat Çağlı ◽

İmdat Yüce ◽

Alperen Vural ◽

Hacı Okuducu ◽

...

Keyword(s):

Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Cancer Patients ◽

Laryngeal Cancer ◽

Squamous Cell ◽

Laryngeal Squamous Cell Carcinoma ◽

Immunohistochemical Analysis ◽

Controlled Study ◽

Tissue Samples ◽

H Pylori

Aim. This study aims to investigate the possible role ofH. pylorias a cause of laryngeal squamous cell carcinoma.Method. This controlled study was performed with 31 consecutive laryngeal cancer and 28 cancer-free patients who underwent direct laryngoscopy and biopsy of laryngeal lesions. To document the previousH. pyloriinfection, serological analysis of the antibody titers was done. Immunohistochemical analyses were applied to the tissue samples.Results. Serology was found positive at the 90.3% of the laryngeal cancer patients and 96.4% of the benign group. There were no statistically significant differences between the two groups (P>0.05). Immunohistochemical analysis results were determined as negative at all of the specimens of laryngeal cancer patients and patients with benign lesions.Conclusion. There were no signs of colonization ofH. pyloriin laryngeal tissues of both groups' patients. It is thought that no relationship exists between theH. pyloriinfection and laryngeal squamous cell carcinoma.

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