TRAF6抑制肽对肾癌细胞的抑制性能研究

Inhibitory effect of TRAF6 inhibitory peptide on renal carcinoma cell

目的研究肿瘤坏死因子受体相关因子6(tumor necrosis factor receptor-associated factors 6,TRAF6)抑制肽的载体材料对肾癌细胞特异性识别,及其对肿瘤细胞生长的影响。方法针对TRAF6,采用固相合成法与缩合反应制设计并制备TRAF6抑制肽-硬脂酸双亲分子(DRQIKIWFQNRRMKWKK-octadecanoic acid),利用电喷雾质谱法(ESI-MS)与高效液相色谱法(HPLC)对双亲分子进行了表征。通过研究TRAF6抑制肽-硬脂酸双亲分子对肾癌细胞(786-O)与正常肾细胞(AD293)的存活率的影响,来评价其对肾癌细胞特异性的识别作用以及对正常细胞/组织的影响。结果针对TRAF6合成了TRAF6抑制肽-硬脂酸双亲分子(DRQIKIWFQNRRMKWKK-octadecanoic acid),ESI-MS测得其相对分子质量为2 628.35,在658.2、877.1和1 315.2处的吸收峰分别为[M+4H]/4、[M+3H]/3和[M+2H]/2的离子峰,HPLC结果显示纯度可达95.84%,均可证明TRAF6抑制肽-硬脂酸双亲分子已成功制备。经过48h的共培养,TRAF6抑制肽-硬脂酸使肾癌细胞存活率下降到74.00%;对正常肾细胞却无明显影响,其细胞存活率仍>90%。结论 TRAF6抑制肽-硬脂酸能被肾癌细胞吞噬,并抑制肿瘤细胞生长,从而体现出一定的靶向性。

OBJECTIVE To investigate the targeting and inhibitory effect of TRAF6 mimicking peptide amphiphiles on tumor growth of renal cell carcinoma.METHODS Solid phase peptide synthesis was used to synthesize the TRAF6 inhibitory peptide and it was further conjugated with octadecanoic acid via condensation reaction.ESI-MS and HPLC was used to characterize the chamical structure of the TRAF6 inhibitory peptide-octadecanoic acid.The prepared TRAF6 inhibitory peptide-octadecanoic acid was introduced to study inbibitory effect on the viability of renal cell.RESULTS Result from ESI-MS indicated the molecular weight of peptide amphiphile was 2 628.35 and the peak at 658.2,877.1and1 315.2could be assigned to absorbance of [M ＋4H]/4,[M ＋3H]/3and [M ＋2H]/2,respectively.HPLC data showed that the content of peptide amphiphile in the product could reach about 95.84%,demonstrating that the peptide amphiphiles has been fabricated successfully.After 48 hincubation,the viability of renal cell carcinoma was decreased to74.00% after incubated with TRAF6 inhibitory peptide-octadecanoic acid with slight kidney cell changes.CONCLUSIONS TRAF6 inhibitory peptide-octadecanoic acid can be selectively absorbed by renal cell carcinoma and has the ability to inhibit growth and proliferation of renal cell carcinoma.