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Published on: May 10, 2022
Functional interleukin 4 receptor and interleukin 2 receptor common gamma-chain on human non-small cell lung cancers:
1Department of Molecular Oncology, John Wayne Cancer Institute at Saint John's Health Center, Santa Monica, CA, USA. essnerr@jwci.org
Objective:
The interleukin 4 receptor has been demonstrated on the surface of human non-small cell lung carcinoma cell lines and tumor specimens. Interleukin 4 causes G1-phase cell-cycle arrest of non-small cell lung cancer cell lines expressing the interleukin 4 receptor; the effect directly correlates with the expression of the interleukin 4 receptor and is seen within 48 hours after treatment. We examined signal transduction pathways used by the interleukin 4 receptor that may account for growth arrest of the cell line LUst but had no effect on another non-small cell lung cancer cell line, SK-MES-1.
Methods:
Western blot analysis was performed on both LUst and SK-MES-1 cell lines cultured in the presence of interleukin 4 (500 U/mL). Cells were lysed, protein extracted, and electroblotted; blots were then probed with murine monoclonal antibodies to specific intracellular proteins.
Results:
Western blotting of the cell lines with antiphosphotyrosine antibody (4G10) demonstrated multiple (140 kd, 100-130 kd, and 65 kd) phosphoproteins seen only in the interleukin 4-treated LUst cell line and not observed in the SK-MES-1 cell lines. Immunoprecipitation and blotting of the LUst cell line with specific secondary antibodies demonstrated that the 140-kd phosphoprotein was the interleukin 4 receptor, the 130-kd phosphoprotein was Janus kinase 1, the 116-kd phosphoprotein was Janus kinase 3, and the 65-kd phosphoprotein was the interleukin 2 receptor gamma-chain. Specific binding was not observed in the non-small cell lung cancer cell line SK-MES-1, suggesting that a functional interleukin receptor gamma-chain was not present. Southern blotting with complementary DNA probes to interleukin 2 receptor gamma-chain confirmed the absence of this receptor on cell line SK-MES-1.
Conclusions:
These results suggest that non-small cell lung cancer cells may express functional cytokine receptors, including the interleukin 2 receptor gamma-chain commonly found in association with the lymphocyte interleukin 2 receptor. These receptors may be novel targets for directing cytokine-based immune therapy.
Insights
Interleukin 4 (IL-4) receptor signaling in non-small cell lung cancer (NSCLC) cells involves the IL-2 receptor gamma-chain. This pathway may offer novel targets for cytokine-based immunotherapy in NSCLC.
Area of Science:
- Molecular Biology
- Cancer Research
- Immunology
Background:
- The interleukin 4 receptor (IL-4R) is present on human non-small cell lung carcinoma (NSCLC) cells.
- IL-4 induces G1-phase cell-cycle arrest in IL-4R-expressing NSCLC cells within 48 hours.
- The specific signal transduction pathways mediating IL-4R effects in NSCLC remain to be fully elucidated.
Purpose of the Study:
- To investigate the signal transduction pathways utilized by the IL-4 receptor in NSCLC cell lines.
- To identify key molecular components involved in IL-4-induced growth arrest.
- To compare signaling in an IL-4-responsive (LUst) versus an IL-4-unresponsive (SK-MES-1) NSCLC cell line.
Main Methods:
- Western blot analysis was performed on LUst and SK-MES-1 cells treated with IL-4.
- Cells were lysed, proteins extracted, and probed with specific monoclonal antibodies.
- Immunoprecipitation and Southern blotting were used to confirm protein presence and absence.
Main Results:
- Western blotting revealed multiple phosphoproteins (140 kDa, 100-130 kDa, 65 kDa) in IL-4-treated LUst cells, but not in SK-MES-1 cells.
- The 140 kDa phosphoprotein was identified as the IL-4 receptor, 130 kDa as Janus kinase 1 (JAK1), 116 kDa as Janus kinase 3 (JAK3), and 65 kDa as the interleukin 2 receptor gamma-chain (IL-2Rγ).
- Southern blotting confirmed the absence of a functional IL-2Rγ in the SK-MES-1 cell line.
Conclusions:
- NSCLC cells can express functional cytokine receptors, including the IL-2Rγ, typically associated with lymphocyte IL-2 receptors.
- The IL-4R signaling pathway in NSCLC involves components like JAK1, JAK3, and IL-2Rγ.
- These identified receptors represent potential novel targets for cytokine-based immunotherapy in NSCLC.
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