CAJ | 학술논문

目的对前期蛋白质组学筛查、鉴定的胰腺癌相关免疫原性候选膜抗原进行验证.方法应用基质辅助激光解吸离子化飞行时间质谱分析与肽质指纹库对前期从66例胰腺癌患者血清的IgG与人胰腺癌细胞株SW1990的膜蛋白免疫印迹杂交的3号和8号阳性点进行鉴定.再应用RT-PCR、real-time PCR和Western blot方法,在细胞株层面从基因及蛋白水平对筛查出的膜抗原进行有效性验证,并比较不同胰腺癌细胞株中目的膜抗原的基因及蛋白表达水平的差异.结果 3号和8号阳性点经质谱分析鉴定分别为电压依赖性离子通道3(VDAC3)和儿茶酚胺氧位甲基转移酶(COMT).经RT-PCR、real-time PCR和Western blot实验证明,候选膜抗原VDAC3和COMT的基因及蛋白在胰腺癌细胞株SW1990、AsPc、P3中均有表达.结论人胰腺癌细胞膜蛋白VDAC3、COMT可能是候选的具有免疫原性的胰腺癌相关膜抗原,并且其基因和蛋白在胰腺癌细胞株SW1990、AsPc、P3中均有表达.
목적 대전기단백질조학사사、감정적이선암상관면역원성후선막항원진행험증.방법 응용기질보조격광해흡리자화비행시간질보분석여태질지문고대전기종66례이선암환자혈청적IgG여인이선암세포주SW1990적막단백면역인적잡교적3호화8호양성점진행감정.재응용RT-PCR、real-time PCR화Western blot방법,재세포주층면종기인급단백수평대사사출적막항원진행유효성험증,병비교불동이선암세포주중목적막항원적기인급단백표체수평적차이.결과 3호화8호양성점경질보분석감정분별위전압의뢰성리자통도3(VDAC3)화인다분알양위갑기전이매(COMT).경RT-PCR、real-time PCR화Western blot실험증명,후선막항원VDAC3화COMT적기인급단백재이선암세포주SW1990、AsPc、P3중균유표체.결론 인이선암세포막단백VDAC3、COMT가능시후선적구유면역원성적이선암상관막항원,병차기기인화단백재이선암세포주SW1990、AsPc、P3중균유표체.
Objective To verify the obtained immunogenic membrane antigens candidate of pancreatic cancer in the performed research. Methods Pancreatic caner cell line SW1990 membrane protein underwent immunoblot with serum IgG purified from clinically collected sera of 66 pancreatic cancer patients. Number 3 and number 8 positive dots of immunoblot were identified by MALDI-TOF mass spectrometry and peptide mass frigerfrint matching. The candidate membrane antigens were further validated in cell lines by RT-PCR, real-time PCR and Western blot, and their different expression level of gene and protein in pancreatic caner cell lines were contrastly studied. Results Number 3 and number 8 positive dots were identified as: voltage-dependent anion channel ( VDAC3 ) and catechol-o-methyltransferase (COMT).RT-PCR,real-time PCR and Western blot showed that gene and protein of VDAC3 and COMT were expressed in the pancreatic cancer cell line SW1990, AsPc and P3 respectively. Conclusion VDAC3 and COMT might be the candidate immunogenic membrane antigens of human pancreatic cancer, and their gene and protein are differently expressed in the pancreatic cancer cell line SW1990,AsPc and P3.