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Updated: Aug 6, 2026
Isolation and Characterization of a Head and Neck Squamous Cell Carcinoma Subpopulation Having Stem Cell Characteristics
Published on: May 11, 2016
Growth stimulation of human head and neck squamous cell carcinoma cell lines by interleukin 4
J N Myers1, S Yasumura, Y Suminami
1Departments of Otolaryngology, Pathology, Molecular Genetics and Biochemistry, University of Pittsburgh, Pennsylvania 15213, USA.
Abstract:
Interleukin 4 (IL-4) has been reported recently to inhibit growth of acute lymphoblastic lymphoma, non-Hodgkin's lymphoma, melanoma, sarcoma, breast, gastric, colon, and renal tumor cell lines, and treatment of murine tumors with IL-4 gene-transduced cells has been therapeutically successful. Therefore, we sought to determine the effect of IL-4 on the growth of human squamous cell carcinoma of the head and neck (SCCHN) cell lines. Growth of SCCHN cell lines incubated in the presence of various concentrations of IL-4 was measured in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide colorimetric assays and by cell counts. Specific binding of IL-4 to SCCHN cells was demonstrated by flow cytometry with phycoerythrin-labeled IL-4, blocking studies with antibodies to IL-4, and using the radiolabeled ligand 125I-labeled IL-4. Reverse transcription PCR for IL-4 and IL-4 receptor (IL-4R) mRNA was performed. SCCHN tissue biopsies were examined by immunohistology and in situ hybridization for the presence of IL-4 protein and IL-4 mRNA in the tumor, respectively. In contrast to earlier reports, we observed growth stimulatory effects of IL-4 consistently in 6 of 13 SCCHN cell lines tested. Growth stimulation by IL-4 ranged from 20 to 200% of control (P < 0.05) and was IL-4 dose dependent. The growth-promoting effect of IL-4 was inhibited completely by incubation of tumor cells in the presence of antibodies specific for IL-4. Reverse transcription PCR analysis of mRNA obtained from the SCCHN cell lines and ELISA performed with SCCHN cell supernatants respectively indicated that the tumor cells did not transcribe or secrete IL-4 actively. The SCCHN cell lines expressed 260-540 IL-4Rs/cell with a dissociation constant of 100 +/- 8 pM. SCCHN cell lines also contained IL-4R mRNA. Immunostaining of SCCHN tissue biopsies indicated that IL-4 may be produced and secreted within these tumors by tumor-infiltrating lymphocytes. In situ hybridization for IL-4 mRNA indicated the presence of positive cells in the tumor stroma. Our data suggest that IL-4 may regulate the growth of SCCHN cells by a paracrine mechanism. These data also indicate that immunotherapy with exogenous IL-4 or IL-4 gene therapy to treat head and neck cancer may not be effective, given the potential tumor growth-stimulatory effects of this cytokine.
Insights
Interleukin 4 (IL-4) unexpectedly stimulated growth in human head and neck squamous cell carcinoma (SCCHN) cell lines, contradicting previous findings. This suggests IL-4 immunotherapy may be ineffective for head and neck cancers.
Area of Science:
- Oncology
- Immunology
- Molecular Biology
Background:
- Interleukin 4 (IL-4) has demonstrated inhibitory effects on various cancer cell lines, including lymphomas, melanomas, and sarcomas.
- Previous research indicated successful therapeutic outcomes in murine tumor models treated with IL-4 gene-transduced cells.
- The role of IL-4 in human squamous cell carcinoma of the head and neck (SCCHN) remained largely unexplored.
Purpose of the Study:
- To investigate the effect of Interleukin 4 (IL-4) on the growth of human squamous cell carcinoma of the head and neck (SCCHN) cell lines.
- To determine the mechanism of IL-4 action, including receptor binding and endogenous production in SCCHN.
- To assess the potential efficacy of IL-4-based therapies for head and neck cancers.
Main Methods:
- SCCHN cell lines were cultured with varying concentrations of IL-4, and growth was assessed using MTT assays and direct cell counts.
- IL-4 binding to SCCHN cells was confirmed via flow cytometry and radioligand assays.
- Reverse transcription PCR (RT-PCR), immunohistology, and in situ hybridization were employed to analyze IL-4 and IL-4 receptor (IL-4R) expression in cell lines and tumor tissues.
Main Results:
- Contrary to prior reports, IL-4 significantly stimulated SCCHN cell growth in 6 out of 13 tested cell lines, with dose-dependent effects ranging from 20% to 200% increase.
- The growth-promoting effect was abrogated by IL-4-specific antibodies, confirming IL-4's direct role.
- SCCHN cells expressed IL-4 receptors (IL-4Rs) and IL-4R mRNA, but did not actively produce or secrete IL-4; IL-4 likely originated from tumor-infiltrating lymphocytes (TILs) via a paracrine mechanism.
Conclusions:
- Interleukin 4 (IL-4) exhibits a growth-stimulatory effect on a subset of human squamous cell carcinoma of the head and neck (SCCHN) cell lines.
- The observed paracrine mechanism, involving IL-4 produced by tumor-infiltrating lymphocytes, suggests a complex role in SCCHN tumor microenvironment.
- These findings indicate that IL-4 immunotherapy or gene therapy may not be an effective treatment strategy for head and neck cancers due to potential tumor growth promotion.
