CAJ | 학술논문

目的对前期蛋白质组学筛查、鉴定的人胰腺癌相关免疫原性候选膜抗原进行有效验证.方法前期实验中,将胰腺癌患者血清提取的IgG与人胰腺癌细胞株SW1990的膜蛋白进行免疫印迹杂交,共获得了9个阳性蛋白位点,现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5号和6号位点进行鉴定.应用RT-PCR、real-time PCR验证二者在不同的人胰腺癌细胞株中的基因表达水平.分别提取人正常胰腺组织和胰腺癌组织的mRNA,应用real-time PCR检测prohibitin 2在人正常胰腺组织中和胰腺癌组织中的基因表达差异.结果 5号和6号阳性蛋白位点经质谱分析鉴定分别是prohibitin 2和prohibitin.RT-PCR和real-time PCR证明,候选膜抗原prohibitin 2和prohibitin在人胰腺癌细胞株SW1990、AsPc、P3的细胞中均有明确的基因表达,尤其在P3细胞中mRNA表达水平最高(t=7.442,P＜0.01).并且prohibitin 2在人胰腺癌组织中的基因表达水平明显高于正常胰腺组织(t=0.893,P＜0.01).结论人胰腺癌细胞膜蛋白prohibitin 2和prohibitin在人胰腺癌细胞株SW1990、AsPc、P3中均有明确的基因表达,并且prohibitin 2在人胰腺癌组织中的基因表达水平明显高于人正常胰腺组织,二者可能是具有免疫原性的胰腺癌相关膜抗原.
목적 대전기단백질조학사사、감정적인이선암상관면역원성후선막항원진행유효험증.방법 전기실험중,장이선암환자혈청제취적IgG여인이선암세포주SW1990적막단백진행면역인적잡교,공획득료9개양성단백위점,현응용기질보조격광해흡리자화비행시간질보분석여태질지문고대5호화6호위점진행감정.응용RT-PCR、real-time PCR험증이자재불동적인이선암세포주중적기인표체수평.분별제취인정상이선조직화이선암조직적mRNA,응용real-time PCR검측prohibitin 2재인정상이선조직중화이선암조직중적기인표체차이.결과 5호화6호양성단백위점경질보분석감정분별시prohibitin 2화prohibitin.RT-PCR화real-time PCR증명,후선막항원prohibitin 2화prohibitin재인이선암세포주SW1990、AsPc、P3적세포중균유명학적기인표체,우기재P3세포중mRNA표체수평최고(t=7.442,P＜0.01).병차prohibitin 2재인이선암조직중적기인표체수평명현고우정상이선조직(t=0.893,P＜0.01).결론 인이선암세포막단백prohibitin 2화prohibitin재인이선암세포주SW1990、AsPc、P3중균유명학적기인표체,병차prohibitin 2재인이선암조직중적기인표체수평명현고우인정상이선조직,이자가능시구유면역원성적이선암상관막항원.
Objective To validate those obtained immunogenic membrane antigens candidate of human pancreatic cancer in the performed research.Methods In the pre-studies,serum IgG purified from clinically collected sera of pancreatic cancer patients underwent immunoblot with human pancreatic caner cell line SW1990 membrane protein,totally obtained 9 positive protein spots.Number 5 and 6 positive dots of immunoblot were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry and peptide mass fingerprinting matching.The candidate membrane antigens were further validated in cell lines by RT-PCR and real-time PCR.RNA of human normal pancreatic tissue and pancreatic cancer tissue was extracted respectively,different gene expression level of prohibitin 2 was studied by real-time PCR.Results Number 5 and 6 positive dots were identified as prohibitin 2 and prohibitin.RT-PCR and real-time PCR all showed that gene of prohibitin 2 and prohibitin were expressed in the human pancreatic cancer cell line SW1990,AsPc and P3 respectively,especially in P3 cell with highest expression( t =7.442,P ＜ 0.01 ).In addition,gene expression level of prohibitin 2 was significant higher in human pancreatic cancer than that of normal pancreatic tissue ( t =0.893,P ＜ 0.01 ). Conclusions Prohibitin 2 and prohibitin are both differently expressed in the pancreatic cancer cell line SW1990,AsPc and P3.Prohibitin 2 is obvious highly expressed in human pancreatic cancer tissue. Prohibitin 2 and prohibitin might be the candidate immunogenic membrane antigens of human pancreatic cancer.