1.Modified serum-guided immunoblotting for differential proteomic study of prostate cancer.
Xiao-bo ZHANG ; Zheng-yan TANG ; Xiong-bin ZU ; Lin QI ; Jing-de RUAN
National Journal of Andrology 2010;16(5):438-444
OBJECTIVETo search for a new method of screening for molecular targets for androgen-dependent prostate cancer.
METHODSWe collected tissue samples and paired serum samples from 3 cases of androgen-dependent prostate cancer (ADPC) treated by surgical resection, and included another 3 samples of benign prostatic hyperplasia (BPH) tissue and normal human serum in the control group. The total proteins extracted were separated and transmembrane by two-dimensional gel electrophoresis, followed by hybridization with the sera of the patients with ADPC and those with hormone-independent prostate cancer (HIPC) as the primary antibodies. The differentially expressed proteins were compared by Western blot, analyzed by MALDI-TOF-MS mass spectrography, and verified by RT-PCR and Western blot following bioinformatic identification.
RESULTSThis modified method exhibited a significantly better effect in displaying differentially expressed proteins, by which 12 differentially expressed protein spots were identified, including Beclin1, glutathione S-transferase P (GSTP1-1), ZBTB7, dihydrodiol dehydrogenase 2 (DDH), enolase (ENO1), glucose-dependent insulin-releasing peptide receptor (GIPR), Mn-superoxide dismutase (MnSOD), phosphoglycerate mutase 1 (PGAM1), amino-peptidyl-prolyl cistrons isomerase (PPIA), and phospholipid-PE-binding protein (PEBP). The mRNA and protein expressions of Beclin1 were significantly down-regulated in androgen-dependent prostate cancer tissues.
CONCLUSIONThis modified serum-guided immunoblotting technique has provided a new method for clarifying the molecular mechanisms of the occurrence and progression of HIPC, in which Beclin1-mediated autophagy may play a key role.
Biomarkers, Tumor ; blood ; Blotting, Western ; Humans ; Immunoblotting ; methods ; Male ; Mass Spectrometry ; Prostatic Neoplasms ; genetics ; metabolism ; Proteomics
2.Expression and clinical significance of B7-H3 and IL-21 in serum of patients with HBV associated hepatocellular carcinoma
Cheng-De LI ; Yi QUAN ; Jian-Mei CAI ; Xiao-Wen WU ; Xue-Ji SHEN ; De-Bin RUAN ; Lin ZHANG ; Hai-Zhu LI
Chinese Journal of Immunology 2018;34(4):569-575
Objective:To investigate the expression and clinical significance of B7-H3 and IL-21 in serum of patients with HBV-related primary liver cancer.Methods: Gathering 121 cases of HBV-related patients,50 cases of primary hepatic carcinoma of them were considered as hepatic carcinoma group,71 cases of benign group including 12 cases with acute hepatitis,21 cases of chronic moderate to severe hepatitis,20 cases of compensatory phase cirrhosis,18 cases of decompensated cirrhosis and 20 cases of healthy persons in the same period as normal control.The content of serum B7-H3 and IL-21 were detected by ELISA.HBV DNA quantitative results were analyzed by Quantitative Real-time PCR.Results: The levels of B7-H3 and IL-21 in patients with primary hepatic carcinoma were (207.60±57.16)ng/ml and(2 357.28±805.01)pg/ml,respectively,which were significantly higher than those of the normal control subjects(P<0.001).Comparison with the normal control subjects,the content of B7-H3 and IL-21 in serum of patients with different clinical types in the benign group increased significantly(P<0.001).B7-H3 and IL-21 were positively associated with each other in serum of patients with HBV-related primary liver cancer.The expression of sB7-H3 was not significantly correlated with the degree of HBV DNA replication.The expression of IL-21 was correlated with HBV DNA replication in patients with HBV associated hepatocellular carcinoma,but was not significantly correlated with the degree of HBV DNA replication.Conclusion: HBV-related primary hepatic carcinoma express sB7-H3 and IL-21 with high level.The continuous high expression of sB7-H3 and IL-21 in the body may be related to the development and prognosis of the patients.