Production of immortal squamous cell line from larynx carcinoma

Eugenol (EU), the active ingredient in clove oil, is commonly used as successful therapeutic compound in dentistry due to its antiseptic and anti-inflammatory effects. Recent research studies suggest that eugenol has also a potential anti-cancer effect. This study was thereby purported to assess the effects of EU on the bioenergetic profile of the SCC-4 human squamous cell carcinoma cell line. To this aim, SCC-4 cells were treated for 24 hours with free EU and EU incorporated in polyurethane structures (50 �M each). Oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were measured using the Seahorse XF-24e extracellular flux analyzer (Agilent Technologies Inc.). Analysis of the SCC-4 bioenergetic profile was performed in the presence of the classic modulators of the electron transport chain: oligomycin, FCCP, and antimycin A+rotenone. Our data showed that cells stimulated with free EU induced a decrease of OCR linked parameters and an increase of ECAR, effects that were abolished by the incorporation of EU in polyurethane structures. In conclusion, free eugenol elicits inhibitory effects on mitochondrial respiration in the SCC-4 cell line, a result that might be suggestive for its anti-tumoral effects.

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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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