The Expression Of ZIP5Gene In Esophageal Cancer And Construction Of Stable Low ZIP5Genes Expressed Esophageal Cancer Cell Line

Objective:1To describe the expression situation of ZIP5gene in esophageal cancertissues, para-carcinoma tissue and normal tissue and to explore the possiblerole and mechanism of ZIP5in the development of esophageal cancer.2To establish the stable esophageal cancer cell line with the lowexpression of ZIP5gene to provide a new theoretical basis for the preventionand targeted therapy of esophageal cancer.Method:1To obtain30specimens of esophageal cancer tissues, para-carcinomatissue and normal tissue, respectively; to contrast their protein expression levelof ZIP5gene detected by immunohistochemical and Western Blot, and tocontrast their mRNA expression level by qRT-PCR.2To choose specific siRNA with the effect of knocking down ZIP5gene,clone it into the plasmid PSD31, then get the stable cell line with low ZIP5gene expression. To check the silence effects by qRT-PCR and Western Blot.Result:1Immunohistochemical result showed that, there was28cases in30esophageal cancer tissues with the expression of ZIP5gene,13cases in30para-carcinoma tissues and3tissues in30normal tissues. The score was4.10±0.51,0.78±0.97,0.20±0.03, respectively. ZIP5gene was expressed in theesophageal cancer tissue, para-carcinoma tissue, normal tissue from higherlevel to lower level in order. Western Blot showed that the protein expressiverelative value of ZIP5was3.05±0.20in cancer tissue,1.78±0.92inpara-carcinoma tissue,0.52±0.29in normal tissue. ZIP5gene was expressedin the esophageal cancer tissue, para-carcinoma tissue, normal tissue from higher level to lower level in order. It was the same as theim-munohistochemical results; qRT-PCR showed that the relative value ofZIP5was14.83±4.63in cancer tissue,4.17±0.79in para-carcinoma tissuebased on the standard normal tissue which was1. ZIP5gene was expressed inthe esophageal cancer tissue, para-carcinoma tissue, normal tissue from higherlevel to lower level in order.2The specific sequences were cloned into TA vector plasmid successfully.The recombinant plasmid was identified by restriction and sequencing.PSD31-ZIP5-shRNA packaged lentivirus and established the stable cell linesuccessfully. qRT-PCR showed that the expression of ZIP5gene inKYSE170K cells was significantly low compared to the KYSE170S cells.Western Blot showed the same result. Stability lower ZIP5geneticexpressionof esophageal cancer cells line was successfully established.Conclusions:1ZIP5gene was expressed in the esophageal cancer tissue,para-carcinoma tissue, normal tissue from higher level to lower level in order.2The stable esophageal cancer cell line with the low expression of ZIP5gene was established.