CAJ | 학술논문

以培养的原发性肝癌细胞HepG2和正常肝细胞L02为研究对象,采用细胞裂解液提取总蛋白,用PNGase F酶解释放N-糖链,以微晶纤维素柱结合石墨碳柱纯化分离N-糖链,通过电喷雾电离质谱( ESI-MS)和串联质谱( MS/MS)对N-糖链进行序列鉴定,以β-环糊精为内标对2种细胞系的N-糖链进行了定量比较分析.结果表明,在肝癌细胞系HepG2和正常细胞系L02中共检测到26种N-糖链,与L02相比, HepG2的大多数高甘露糖型糖链、唾液酸化糖链和岩藻糖基化糖链的数量都明显升高,其中有15种糖链在数量上具有极显著性差异(p<0.01),1种糖链具有显著性差异(p<0.05).本研究为进一步探索肝癌中各类N-糖链的表达特点及发现早期肝癌糖链标志物提供了参考.
이배양적원발성간암세포HepG2화정상간세포L02위연구대상,채용세포렬해액제취총단백,용PNGase F매해석방N-당련,이미정섬유소주결합석묵탄주순화분리N-당련,통과전분무전리질보( ESI-MS)화천련질보( MS/MS)대N-당련진행서렬감정,이β-배호정위내표대2충세포계적N-당련진행료정량비교분석.결과표명,재간암세포계HepG2화정상세포계L02중공검측도26충N-당련,여L02상비, HepG2적대다수고감로당형당련、타액산화당련화암조당기화당련적수량도명현승고,기중유15충당련재수량상구유겁현저성차이(p<0.01),1충당련구유현저성차이(p<0.05).본연구위진일보탐색간암중각류N-당련적표체특점급발현조기간암당련표지물제공료삼고.
HepG2, a primary hepatocellular carcinoma cell line, and L02 derived from normal liver tissue, were chosen as model cell lines. In this work, the total proteins of HepG2 and L02 cells were extracted and enzymatically hydrolyzed by Peptide-N-glycosidase F( PNGase F) . Then the released N-glycans were purified by microcrystalline cellulose and graphitized carbon columns. The structure of the purified N-glycans was iden-tified by electrospray ionization mass spectrometry( ESI-MS) and MS/MS. To compare the difference of N-gly-cans between HepG2 and L02 cells, β-cyclodextrin was used as the internal standard to relative quantitative analysis of the N-glycans by MS. As results, 26 N-glycans are observed in both HepG2 and L02 cells. The amounts of high-mannose, sialylated, as well as fucosylated and sialylated N-glycans of HepG2 were higher than those of L02 cells. Among them, 15 kinds of N-glycans of HepG2 cells show very significant difference (p<0.01) and 1 kind of N-glycan show significant difference(p<0.05), compared to those of L02 cells. Our studies show potential in investigation of structure partterns of N-glycans expressed in hepatocellular carcinoma and in finding early biomarker in liver cancer diagnose.