Overexpression of Interleukin-18 Induces Growth Inhibition, Apoptosis and Gene Expression Changes in a Human Tongue Squamous Cell Carcinoma Cell Line
Weijiang Liu, Bing Han, Baozhen Sun, Yan Gao, Yinghui Huang, Min Hu
Abstract
Weijiang Liu, Bing Han, Baozhen Sun, Yan Gao, Yinghui Huang, Min Hu
Abstract
OBJECTIVE: To investigate the role of interleukin-18 (IL-18) in regulating the growth of the human tongue squamous cell carcinoma cell line CRL-1623™. METHODS: The human IL18 gene was cloned and transfected into CRL-1623™ cells using the transfection vector pcDNA3.1(+). Investigations included analysis of cell viability, detection of apoptosis using annexin V-fluorescein isothiocyanate, assessment of caspase 3/7 activity and real-time reverse transcription-polymerase chain reaction to assess expression of the IL18, CCND1 (cyclin D(1)), CCNA1 (cyclin A(1)) and IFNG (interferon-γ) genes. RESULTS: Introduction of the IL18 gene inhibited cell proliferation at 24, 48 and 72 h after transfection compared with untransfected cells and cells transfected with blank pcDNA3.1(+) vector. Apoptotic cell numbers and caspase 3/7 activity were significantly enhanced by IL18 transfection. Levels of IL18 and IFNG mRNA were elevated and CCND1 mRNA was reduced after 48 h in IL18 transfected cells compared with wild-type cells. CONCLUSIONS: These findings suggest that IL-18 plays a role in the regulation of tongue squamous cell carcinoma and may represent a potential therapeutic target.
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OBJECTIVE: To investigate the role of interleukin-18 (IL-18) in regulating the growth of the human tongue squamous cell carcinoma cell line CRL-1623™. METHODS: The human IL18 gene was cloned and transfected into CRL-1623™ cells using the transfection vector pcDNA3.1(+). Investigations included analysis of cell viability, detection of apoptosis using annexin V-fluorescein isothiocyanate, assessment of caspase 3/7 activity and real-time reverse transcription-polymerase chain reaction to assess expression of the IL18, CCND1 (cyclin D(1)), CCNA1 (cyclin A(1)) and IFNG (interferon-γ) genes. RESULTS: Introduction of the IL18 gene inhibited cell proliferation at 24, 48 and 72 h after transfection compared with untransfected cells and cells transfected with blank pcDNA3.1(+) vector. Apoptotic cell numbers and caspase 3/7 activity were significantly enhanced by IL18 transfection. Levels of IL18 and IFNG mRNA were elevated and CCND1 mRNA was reduced after 48 h in IL18 transfected cells compared with wild-type cells. CONCLUSIONS: These findings suggest that IL-18 plays a role in the regulation of tongue squamous cell carcinoma and may represent a potential therapeutic target.
Key concepts: Transfection, Molecular biology, Squamous carcinoma, Cyclin D1, Cell culture, Apoptosis, Cell growth, Cancer research