应用iTRAQ技术筛选乳腺癌白苔和黄苔患者血清差异表达蛋白

Screening Serum Differential Protein with iTRAQ Technology in Breast Cancer for the Patients with White or Yellow Tongue Coating

目的应用同位素标记相对和绝对定量(iTRAQ)技术筛选乳腺癌白苔和黄苔患者与正常组之间的血清差异表达蛋白,以寻找乳腺癌早期诊断的生物标记物和探索中医舌苔形成的机制和原理。方法临床收集乳腺癌患者中具有典型的白厚苔和黄厚苔者各10例,并以正常薄白苔者10例作为对照,采血取血清提取蛋白,然后进行蛋白变性、还原及酶解,最后进行iTRAQ标记和ESI-QTOF-QⅡ质谱仪鉴定,得到峰图用DATA ANALYSIS、MASCOTT 2.2搜库和Scaffold软件分析,得到表达量差异结果。结果共鉴定出335个蛋白,其中达到严格定量标准的蛋白数59个,与正常组比较,乳腺癌白苔组和黄苔组共筛选出11个血清差异表达蛋白,乳腺癌白苔组和黄苔组之间筛选出6个差异蛋白。结论 iTRAQ结合LC-MS/MS技术能够快速、有效地进行差异蛋白质组学研究,同时发现一些蛋白与乳腺癌的发生以及中医舌苔形成机制密切相关。

Objective To screen the serum differential expression protein in breast cancer between the patients with white or yellow tongue coating and normal group so as to seek after the biomarker for the early diagnosis of breast cancer and explore the mechanism of tongue coating formation in traditional Chinese medicine.Methods The patients with typical white thick tongue coating or yellow thick tongue coating were collected from the clinic among the cases of breast cancer,10 cases in each group.Additionally,the people with normal thin white tongue coating were collected as a control group.Blood was collected and protein was extracted from serum.Afterwards,albuminous degeneration,reduction and enzymolysis were performed.Finally,iTRAQ was adopted for labeling and ESI-QTOF-QⅡ mass spectrometer was for identification.The peak figures obtained were searched with DATA AANALYSIS and MASCOTT2.2 and analyzed with Scaffold software so as to achieve the expression difference.Results Totally,335 proteins were identified.Of them,59 proteins reached the strict quantitative standard.Compared with normal group,11 serum differential expression proteins were screened totally from white tongue coating group and yellow tongue coating group for the patients with breast cancer.6 differential proteins were screened in comparison between white tongue coating group and yellow tongue coating group.Conclusion The combination of iTRAQ and LC-MS/MS technology can quickly and effectively perform the study of differential proteomics.Additionally,it is discovered that some proteins are closely relevant with the occurrence of breast cancer and the formation of tongue coating in traditional Chinese medicine.