Screening And Verification Of Novel Biomarkers For Serum Of Thyroid Cancer

Objective The development of ultrasound technology has greatly improved the detection rate of thyroid nodules.However,for the determination of the nature of nodules,there are still no biomarkers with non-invasive but high diagnostic efficiency.The clinical classification of Thyroid Cancer(TC)may bring completely different clinical interventions,which in turn affect the prognosis and long-term quality of life.Therefore,it is necessary to screen new biomarkers for early diagnosis and differential diagnosis of TC.The aim of our study is to analysis of proteome and metabolome of patients with different types of thyroid cancer,screen for valuable markers of thyroid cancer,and do the verification.Methods 31 patients with TC(stage I-IV):15 patients with Papillary Thyroid Cancer(PTC),10 patients with Medullary Thyroid Cancer(MTC),6 patients with Follicular Thyroid Cancer(FTC),and 15 patients with benign thyroid nodules(TN)and 15 healthy controls(HC)were treated with mass spectrometer Orbitrap Q-Exactive-plus for proteome analysis.Six differentially expressed proteins,Amyloid beta A4 protein,Apolipoprotein A-IV,Gelsolin,Contactin-1,Gamma-Glutamyl hydrolase,Complement factor H-related protein 1(CFHR1)were screened in 223 TC patients by Enzyme-linked immunosorbent assay(ELISA).CFHR1 was further validated by ELISA in 291 patients with TC(stage I-IV)(247 patients with PTC,38 patients with MTC and 6 patients with FTC)and 69 patients with TN and 176 patients with HC.Serum lipid mass spectrometry was selected from November 1st,2013 to November 11th,2018 in Peking Union Medical College Hospital.The pathological results were clearly diagnosed as TN and TC subjects,At the same time,for the health check-up population of Peking Union Medical College Hospital from May 1st,2018 to November 11th,2018,HC was selected for the diagnosis of thyroid-free disease.Finally,3,875 subjects were involved.Analysis of the difference in expression levels of lipid metabolism-related molecular such as cholesterol(CHO),triglycerides(TG),high-density lipoprotein-cholesterol(HDL-C),low-density lipoprotein cholesterol(CDL)Low-density lipoprotein-cholesterol,LDL-C),apolipoprotein A1(ApoAl)and apolipoprotein B(ApoB).Results Seventeen differentially expressed proteins in the cancer group and the control group were screened using mass spectrometry data-independent acquisition techniques(P<0.05 and fold-change ratios2 or ≤0.5).The expression levels of CFHR1 protein in the six differentially identified proteins were significantly different among the groups(P<0.05).Further validation of big sample size showed a significant increase in CFHR1 serum levels in the MTC and FTC groups compared with the control and PTC groups(P<0.001).CFHR1 has a higher differential diagnosis(P<0.001)in distinguishing between MTC patients and PTC patients,with a sensitivity of 100.0%,a specificity of 85.08%,an area under the curve of 0.93,and a diagnostic threshold of 0.92 ng/mL.After combined with calcitonin,the specificity was increased to 98.90%;IPA analysis of the LXR/RXR activation pathway was inhibited,which is the key to lipid metabolism and is the main pathway of involvement in this study.Lipid mass spectrometry results showed that the total CHO levels of the cancer group(PTC,FTC and MTC)and the control group(TN and HC)were significantly different between males and females(P<0.05).In women,there were significant differences in TG,HDL-C,Apo A1,and LDL-C/HDL-C levels between the two groups(all P<0.05).The levels of TG and LDL-C/HDL-C were higher in the cancer group than in the control group,and the levels of Apo A1 and HDL-C in the cancer group were lower than those in the control group.Conclusion CFHR1 has higher sensitivity and specificity in the differential diagnosis of PTC and MTC,and screening results show that the ApoA1,ApoA2 and ApoA4 levels in the cancer group are lower than in the healthy control group.Further studies have found that there are differences in the expression of lipid molecules between the cancer group and the control group.It is necessary to further elucidate the molecular mechanism of the association between lipid metabolism and thyroid cancer to develop novel biomarkers and therapeutic targets.