| Budget Amount *help |
¥3,920,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥420,000)
Fiscal Year 2007: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2006: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Research Abstract |
Co-inhibitory molecules are known to induce immune tolerance. Among these, B7-H1 (PD-L1) is expressed in cancer cells, and may cancel anti-tumor immunity by inducing immune tolerance. However, little is known whether B7-H1 is expressed in head and neck cancers in Japanese population
We investigated the expression and characterization of B7-H1 in head and neck cancers in Japanese population. Immunohistochemical staining revealed that B7-H1 protein is expressed in tongue, oral, pharyngeal and laryngeal cancers. RT-PCR analysis confirmed the expression of this molecule in head and neck cancers at mRNA level. Next, we determined the amounts of B7-Hl mRNA in head and neck cancers by real-time PCR, and analyzed the correlation of the amounts of B7-H1 mRNA and clinical characteristics. No significant correlation was seen between the amounts of B7-H1 mRNA and staging including tumor size, lymph node metastasis, distinct metastasis, recurrence, or survival rate.
In this project, we tried knock-down of B7-H1 in SCCVII cell, the mouse squamous cell carcinoma cell line, using siRNA. Flow cytometric analysis revealed that SCCVII cell expressed B7-Hl. Template DNA for whole transcription of RNA for B7-H1 was made, and dsRNA was generated. Then siRNA was made by fragmentation of dsRNA by Dicer siRNA was transfected into SCCVII. However, decrease in B7-H1 expression in SCCVII was marginal (about 10%). The change of B7-H1 expression was similar when we changes the buffets for transfection from GeneSilencer to siIMPORTER, lipofectamine, and RNAiMAX.
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