Studies Of Plasma Protein Markers Common To Abnormal Savda Type Tumors Based On Breast Cancer

Objectives: According to humor pathology(humoral theory)of Uyghur medicine,Abnormal Savda is the major syndrome and pathological condition of complex diseases,such as tumors,type 2 diabetes mellitus and cardiovascular diseases.In previous studies,our research team identified a profile of 7 differentially expressed proteins common to tumor patients with Abnormal Savda and the effect of Abnormal Savda prescription,but the specificity and accuracy of the protein profile has not been validated.In this study,we will investigate the asscociation of Abnormal Savda type breast cancer with the regulation of candidate protein expression by syndrome diagnosis of breast cancer patients in Uyghur medicine and quantitative determination of candidate protein expression in the plasma and tumor tissuesResults of this study will provide evidence for revealing scientific interpretation of medical theories and clinical practice of Uyghur medicine,such as “same syndrome of different diseases” and “a disease coupled to a syndrome”.Methods:(1)We collected peripherial blood from 60 cases of breast cancer patients and 20 cases of age-matched healthy women as normal controlsPatients were classified as 38 cases of Abnormal Savda type breast cancer and non-Abnormal Savda,which include patients.with other syndromes.Meanwhile,56 cases of breast cancer tissues and 24 cases of normal controls were collected regardless of syndrome types of patients.(2)Based on previous studies and focused on the profile of 6 candidate proteins for Abnormal Savda type tumors,including vWF(von Willebrand factor),THBS1(thrombospondin 1),IGFBP2(insulin-like growth factor-binding protein 2),ORM1(orosomucoid 1),FGA(fibrinogen alpha chain)and SAA1(serum amyloid A-1 protein),we analyzed the protein expression in all periphereal blood(plasma)samples by enzyme-linked immunosorbent assay(ELISA),to reveal the association of Abnormal Savda type breast cancer with the regulation of these proteins.(3)We analyzed the mRNA expression of genes coding for candidate proteins in tissue specimens of breast cancer and normal controls using mRNA-specific primers and quantitative RT-PCR method,to reveal th association of breast cancer with the regulation of candidate proteins in tumor tissues.(4)By Pearson correlation and ROC(Receiver Operating Characteristic)curve analysis,we evaluated the correlation between the expression of candidate proteins and the detection potential of these protein on accuracy,specificity and sensitivity.Results:(1)ELISA analysis demonstrated that the plasma levels of all 6 candidate proteins,SAA1,THBS1,vWF,IGFBP2,ORM1 and FGA,were significantly increased in breast cancer patients with Abnormal Savda compared to those with non-Abnormal Savda(other syndrome types)and normal controls(P<0.05);According to Pearson correlation analysis,the altered protein expression between two proteins,SAA1 and THBS1,and among three proteins vWF,ORM1 and FGA was to some extent correlated;ROC curve analysis indicated that the detection of SAA1,vWF,IGFBP2 and FGA had a realative high accuracy,among which the detection of SAA1 and IGFBP2 had a relatively high specificity and sensitivity.(2)RT-PCR data showed that the transcription levels of SAA1,THBS1,IGFBP2,vWF and ORM1 were significantly decreased in tumor specimens from breast cancer compared with normal controls(P<0.05);The altered expression of three genes,SAA1,vWF and ORM1,and between two genes THBS1 and IGFBP2 was relative high correlated;The detection of SAA1,vWF,THBS1 and IGFBP2 in tumor tissues had a high accuracy,specificity and sensitivity,ORM1 detection had high sensitivity,but low specificity and sensitivity.Conclusions: The findings of this study s?ggest that the development of Abnormal Savda type breast cancer is closely associated with the regulation of candidate protein expression that is common to Abnormal Savda type tumors,and demonstrated that the proteomic data may have relatively high accuracy and feasibility.In contrast to the upregulation of candidate proteins in the plasma during breast carcinogenesis,the transcription leves of corresponding genes might be downregulated in tumor tissues,which has to be further validated in future studies.