STC2 MRNA Expression In Hepatocellular Carcinoma And Monoclonal Antibodies Raised Against It

Stanniocalcin(STC)which first identified in bony fish is a glycoprotein hormone and acts as a regulator of calcium and phosphorus balance.In recent years,it has been found that STC2 plays an important role in various malignant cancer,such as liver cancer,breast cancer and ovarian cancer.Studies have shown that STC2 is overexpressed in a variety of cancer tissues compared to adjacent normal tissues,and the expression of STC2 is highly correlated with histological grade and tumor progression.Regulation of STC2 expression alters cell proliferation and epithelial mesenchymal transformation(EMT).Studies have found that the overall survival rate(OS)of HCC patients with high serum STC2 levels is significantly lower than that of patients with lower serum STC2 levels,suggesting that STC2 is a potential biomarker for the diagnosis and prognosis of HCC,and is superior to AFP in some respects.In this study,bioinformatics was firstly used to analyze the expression level of STC2 in tumor tissues of patients with HCC in TCGA and NCBI GEO databases,and it was found that compared with non-tumor tissues,the expression level of STC2 in HCC tissues was up-regulated.The protean software was used to predict and analyze the STC2 protein antigen determination cluster,and the distribution of STC2 protein antigen determination cluster was obtained.According to the prediction results,the 201-302 amino acid sequences of STC2 protein were selected to construct the STC2-C recombinant protein and to prepare the STC2 monoclonal antibody.We used prokaryotic expression technology to clone the amino acid sequence of STC2 protein 201-032 into several prokaryotic expression vectors and constructed recombinant expression plasmids.The soluble recombinant human STC2-C protein was obtained by optimizing the induction conditions,and the prokaryotic expression plasmid was transformed into the expressed strain for amplification and culture.The recombinant human STC2-C protein was expressed under the induction of IPTG and purified.After the tag was excised by bovine thrombase,Balb/C mice were immunized with the obtained STC2-C protein to obtain multiple strains of hybrid tumor cell lines with high titer and secreted specific antibodies,which laid a foundation for the subsequent development of human STC2 protein double-anti-sandwich ELISA and detection kit.Conclusion:Compared with with normal liver tissues,the expression level of STC2 mRNA in tumor tissues of patients with hepatocellular carcinoma was significantly increased,and the prognosis of patients with high expression of STC2 mRNA was worse.In this study,the Protein module of DNAstar software was used to predict the distribution of STC2 Protein antigen determination cluster.Based on the predicted results,the recombinant human STC2-C Protein was constructed,and a number of hybrid tumor cell lines with high titer and secreted specific antibodies were initially obtained.