2019 Fiscal Year Final Research Report

Analysis of alteration of N-glycan and invasiveness in hepatocellular carcinoma cell-lines

Research Project

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Project/Area Number	16K10561
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	Digestive surgery
Research Institution	Hokkaido University
Principal Investigator	Kamiyama Toshiya 北海道大学, 医学研究院, 准教授 (80322816)
Co-Investigator(Kenkyū-buntansha)	西村紳一郎北海道大学, 先端生命科学研究院, 教授 (00183898) 島田慎吾北海道大学, 医学研究院, 特任助教 (40755576) 若山顕治北海道大学, 医学研究院, 客員研究員 (50646544) 武冨紹信北海道大学, 医学研究院, 教授 (70363364) 横尾英樹旭川医科大学, 医学部, 准教授 (70399947) 折茂達也北海道大学, 医学研究院, 特任助教 (80711861)
Project Period (FY)	2016-04-01 – 2020-03-31
Keywords	糖鎖 / 浸潤 / 転移 / 肝細胞癌
Outline of Final Research Achievements	We investigated the correlation between alterations in N-glycans and invasiveness by u-PA and EMT. Expression of u-PA and E-glycans/N-cadherin were analyzed by Western blotting in hepatocellular carcinoma (HCC) cell lines. u-PA were knocked down by RNA interference and u-PA was overexpressed in HCC cells using lentiviral vectors. We performed a glycoblotting-assisted MALDI-TOF/MS-based quantitative analysis of HCC cell lines in which invasiveness was altered. The expression of N-glycans, including a form with m/z=1851 and 2521, was changed according to invasiveness controlled by knockdown and overexpression of u-PA expression. These two N-glycans were common in the comparative analysis between HLE with low E-Cadherin expression and HepG2 with high expression. Conclusion: In HCC cells, N-glycosylation is an important factor controlling invasiveness by u-PA and EMT related with E-Cadherin and N-Cadherin. These glycomic alteration may be useful for evaluation of tumor malignancy.
Free Research Field	外科　腫瘍学
Academic Significance and Societal Importance of the Research Achievements	肝癌培養細胞を用い、網羅的糖鎖解析を行い、u-PAと上皮間葉転換EMTに関連する浸潤能に関与する特異的糖鎖1851m/z、2521m/zが浸潤能変化に検出できた。これらの糖鎖はu-PA、E-Cadherin、N-Cadherinの発現の変化に伴い変化した。これらから癌悪性度変化に関連する糖鎖が検出され、既存のAFP 、AFP-L3、PIVKA-IIと違った糖鎖という視点から肝細胞癌悪性度を評価できる新規バイオマーカーとなり得、これらの糖鎖に関与する糖転移酵素阻害剤を合成できれば、浸潤制御、上皮間葉転換EMT制御ができ、予後延長の可能性があり、創薬の点からも癌治療に大きく貢献できると確信する。