Radiation Sensitization of Basal Cell and Head and Neck Squamous Cell Carcinoma by the Hedgehog Pathway Inhibitor Vismodegib

Vismodegib, an inhibitor of the Hedgehog signaling pathway, is an approved drug for monotherapy in locally advanced or metastatic basal cell carcinoma (BCC). Data on combined modality treatment by vismodegib and radiation therapy, however, are rare. In the present study, we examined the radiation sensitizing effects of vismodegib by analyzing viability, cell cycle distribution, cell death, DNA damage repair and clonogenic survival in three-dimensional cultures of a BCC and a head and neck squamous cell carcinoma (HNSCC) cell line. We found that vismodegib decreases expression of the Hedgehog target genes glioma-associated oncogene homologue (GLI1) and the inhibitor of apoptosis protein (IAP) Survivin in a cell line- and irradiation-dependent manner, most pronounced in squamous cell carcinoma (SCC) cells. Furthermore, vismodegib significantly reduced proliferation in both cell lines, while additional irradiation only slightly further impacted on viability. Analyses of cell cycle distribution and cell death induction indicated a G1 arrest in BCC and a G2 arrest in HNSCC cells and an increased fraction of cells in SubG1 phase following combined treatment. Moreover, a significant rise in the number of phosphorylated histone-2AX/p53-binding protein 1 (γH2AX/53BP1) foci in vismodegib- and radiation-treated cells was associated with a significant radiosensitization of both cell lines. In summary, these findings indicate that inhibition of the Hedgehog signaling pathway may increase cellular radiation response in BCC and HNSCC cells.

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1,25(OH)2Vitamin D3 induces elevated expression of the cell cycle inhibitor p18 in a squamous cell carcinoma cell line of the head and neck

Journal of Oral Pathology and Medicine ◽

10.1111/j.1600-0714.2006.00407.x ◽

2006 ◽

Vol 35 (8) ◽

pp. 472-478 ◽

Cited By ~ 16

Author(s):

Claudia Gedlicka ◽

Gudrun Hager ◽

Martina Weissenbock ◽

Wilhelm Gedlicka ◽

Birgit Knerer ◽

...

Keyword(s):

Cell Cycle ◽

Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Head And Neck ◽

Cell Line ◽

Squamous Cell ◽

Carcinoma Cell ◽

Squamous Cell Carcinoma Cell ◽

Cell Cycle Inhibitor ◽

Elevated Expression

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Ethanol Decreases Negative Cell-cycle-regulating Proteins in a Head and Neck Squamous Cell Carcinoma Cell Line

Acta Oto-Laryngologica ◽

10.1080/000164802753648277 ◽

2002 ◽

Vol 122 (3) ◽

pp. 338-342 ◽

Cited By ~ 6

Author(s):

Johannes Kornfehl ◽

Gudrun Hager ◽

Claudia Gedlicka ◽

Michael Formanek

Keyword(s):

Cell Cycle ◽

Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Head And Neck ◽

Cell Line ◽

Squamous Cell ◽

Carcinoma Cell ◽

Neck Squamous Cell Carcinoma ◽

Squamous Cell Carcinoma Cell ◽

Negative Cell

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GANT61 Reduces Hedgehog Molecule (GLI1) Expression and Promotes Apoptosis in Metastatic Oral Squamous Cell Carcinoma Cells

International Journal of Molecular Sciences ◽

10.3390/ijms21176076 ◽

2020 ◽

Vol 21 (17) ◽

pp. 6076

Author(s):

Taís Bacelar Sacramento de Araújo ◽

Leonardo de Oliveira Siquara da Rocha ◽

Manuela Torres Andion Vidal ◽

Paulo Lucas Cerqueira Coelho ◽

Mitermayer Galvão dos Reis ◽

...

Keyword(s):

Gene Expression ◽

Cell Cycle ◽

Squamous Cell Carcinoma ◽

Cell Death ◽

Oral Squamous Cell Carcinoma ◽

Cell Viability ◽

Cell Carcinoma ◽

Cell Line ◽

Squamous Cell ◽

Nuclear Fragmentation

Due to its importance in the pathogenesis of oral squamous cell carcinoma (OSCC), the Hedgehog (HH) pathway is considered a potential therapeutic target. We investigated the effects of GANT61, a GLI inhibitor, on HH gene expression, as well as on metastatic OSCC cell proliferation and death. Following culture in DMEM medium, cytotoxicity of GANT61 against different tumor and non-tumor cell types was assessed by alamarBlue assays. Cytotoxicity analysis revealed that the metastatic HSC3 cell line was the most sensitive (IC50: 36 µM) to the tested compound. The compound’s effects on the expression of HH pathways components were analyzed by qPCR and Western blot; cell viability was analyzed by trypan blue assay and flow cytometry were used to investigate cell cycle phase, morphology, and death patterns in HSC3 cells. A significant reduction in mRNA levels of the GLI1 transcription factor was found after 12 h of treatment withGANT61. Protein expression levels of other HH pathway components (PTCH1, SHH, and Gli1) and HSC3 cell viability also decreased after 24 h of treatment. Cell cycle analysis and death pattern evaluations revealed significantly increased nuclear fragmentation in sub-G1 phase, as well as cell death due to apoptosis. In conclusion, the significantly reduced GLI1 gene expression seen in response to the GLI inhibitor indicates diminished downstream activation in HH pathway components. GANT61 significantly reduced cell viability in the metastatic cell line of OSCC and promoted a significant increase in nuclear fragmentation and cell death by apoptosis.

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Genistein-Induced Cell Cycle Arrest and Apoptosis in a Head and Neck Squamous Cell Carcinoma Cell Line

Nutrition and Cancer ◽

10.1207/s15327914nc340102 ◽

1999 ◽

Vol 34 (1) ◽

pp. 12-19 ◽

Cited By ~ 53

Author(s):

Samir A. Alhasan ◽

Haline Pietrasczkiwicz ◽

Maria D. Alonso ◽

John Ensley ◽

Fazlul H. Sarkar

Keyword(s):

Cell Cycle ◽

Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Head And Neck ◽

Cell Cycle Arrest ◽

Cell Line ◽

Squamous Cell ◽

Carcinoma Cell ◽

Squamous Cell Carcinoma Cell ◽

Cycle Arrest

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miR-17-5p drives G2/M-phase accumulation by directly targeting CCNG2 and is related to recurrence of head and neck squamous cell carcinoma

BMC Cancer ◽

10.1186/s12885-021-08812-6 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Qiang Huang ◽

Yu-Jie Shen ◽

Chi-Yao Hsueh ◽

Yang Guo ◽

Yi-Fan Zhang ◽

...

Keyword(s):

Cell Cycle ◽

Flow Cytometry ◽

Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Head And Neck ◽

Cell Lines ◽

Squamous Cell ◽

Target Gene ◽

Cell Cycle Distribution ◽

M Phase

Abstract Background The human miR-17-92 polycistron is the first reported and most well-studied onco-miRNA with a cluster of seven miRNAs. miR-17-5p, a member of the miR-17-92 family, plays an important role in tumor cell proliferation, apoptosis, migration and invasion. However, few studies have shown the role of miR-17-5p in the cell cycle of head and neck squamous cell carcinoma (HNSCC). Methods RT-qPCR was used to detect miR-17-5p expression levels in 64 HNSCC tissues and 5 cell lines. The relationship between the expression of miR-17-5p in the tissues and the clinical characteristics of the patients was analyzed. HNSCC cells were transfected with an miR-17-5p mimic or inhibitor to evaluate cell cycle distribution by flow cytometry. Cell cycle distribution of cells transfected with target gene was evaluated using flow cytometry. Dual-luciferase reporter assay was used to detect the regulatory effect of miR-17-5p on target gene expression. Results In the present study, we found that miR-17-5p expression in HNSCC tissues and cell lines was remarkably increased, and miR-17-5p is related to recurrence in HNSCC patients. Silencing miR-17-5p blocked HNSCC cells in G2/M phase, whereas its overexpression propelled cell cycle progression. More importantly, we verified that miR-17-5p negatively regulated CCNG2 mRNA and protein expression by directly targeting its 3’UTR. Conclusion These findings suggest that miR-17-5p might act as a tumor promoter and prognostic factor for recurrence in HNSCC patients.

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Specific Targeting of HER2-Positive Head and Neck Squamous Cell Carcinoma Line HN5 by Idarubicin-ZHER2 Affibody Conjugate

Current Cancer Drug Targets ◽

10.2174/1568009617666170427105417 ◽

2018 ◽

Vol 19 (1) ◽

pp. 65-73 ◽

Cited By ~ 5

Author(s):

Marzieh Ghanemi ◽

Aminollah Pourshohod ◽

Mohammad Ali Ghaffari ◽

Alireza kheirollah ◽

Mansour Amin ◽

...

Keyword(s):

Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Head And Neck ◽

Cell Line ◽

Squamous Cell ◽

Optimum Concentration ◽

Her2 Positive ◽

Affibody Molecules ◽

Specific Targeting ◽

Mcf 7

Background:Expression of human epidermal growth factor receptor type 2 (HER2) in head and neck squamous cell carcinoma (HNSCC) cell line HN5 can be employed with great opportunities of success for specific targeting of anti-cancer chemotherapeutic agents.Objective:In the current study, HER2-specific affibody molecule, ZHER2:342 (an engineered protein with great affinity for HER2 receptors) was selected for conjugation to idarubicin (an anti-neoplastic antibiotic).Method:ZHER2:342 affibody gene with one added cysteine code at the its 5′ end was synthesized de novo and then inserted into pET302 plasmid and transferred to E. Coli BL21 hosting system. After induction of protein expression, the recombinant ZHER2 affibody molecules were purified using Ni- NTA resin and purity was analyzed through SDS-PAGE. Affinity-purified affibody molecules were conjugated to idarubicin through a heterobifunctional crosslinker, sulfosuccinimidyl 4-(Nmaleimidomethyl) cyclohexane-1-carboxylate (Sulfo-SMCC). Specific toxicity of idarubicin-ZHER2 affibody conjugate against two HER2-positive cells, HN5 and MCF-7 was assessed through MTT assay after an exposure time of 48 hours with different concentrations of conjugate.Results:Idarubicin in the non-conjugated form showed potent toxic effects against both cell lines, while HN5 cells were significantly more sensitive compared to MCF-7 cells. Dimeric ZHER2 affibody showed a mild decreasing effect on growth of both HN5 and MCF-7 cells at optimum concentration. Idarubicin-ZHER2 affibody conjugate at an optimum concentration reduced viability of HN5 cell line more efficiently compared to MCF-7 cell line.In conclusion, idarubicin-ZHER2 affibody conjugate in optimum concentrations can be used for specific targeting and killing of HN5 cells.

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