| Background Ring finger protein 212(RNF212)encodes a loop finger protein that acts as an ubiquitin ligase.The prediction of gene orthologs suggests that the mammalian RNF212 gene has considerable sequence homology with the Caenorhabditis elegans Zhp3 gene.In the cloned murine full-length RNF212 gene,the 307 amino acid protein has an N-terminal loop finger domain followed by a coiled-coil domain and a C-terminal serine-rich domain.The loop finger domain is characterised by a E3-ubiquitin ligase,thereby catalysing protein modification.The human RNF212 protein has significant identity to the full-length mouse protein and has a similar domain structure.The RNF212 gene is located in the human chromosome 4p16.3,and widely expressed in many tissues and organs of the human body.It has different degrees of expression in the stomach,duodenum,colon,oesophagus,liver,gallbladder and appendix.The RNF212 gene plays an important role in cell signalling;cellular assembly and organisation;and DNA replication,recombination and repair.RNF212 is associated with various human tumours,of which RNF212 is associated with poor progression of melanoma because the gene is overexpressed during failure caused by primary melanoma.In addition,the RNF212 gene is associated with male breast cancer in the presence of point mutations or insertional deletion mutations and is lost or deleted in female breast cancer.However,little is known about the carcinogenic effects of the RNF212 gene in GC.The purpose of this study was to further explore the expression of RNF212 in gastric cancer and adjacent non-tumour tissues,and to analyze the relationship between clinical pathological characteristics of patients with abnormal expression of RNF212.The expression differences in protein and m RNA levels of RNF212 in seven gastric cancer cell lines(HGC-27,、MGC-803,、BGC-823,、AGS、SGC-7901、SNU-1 、SNU-16)and human normal gastric mucosa cells(GES-1)were studied,and the effects of RNF212 on the biological functions of gastric cancer cell proliferation and migration were further explored through RNF212 interference plasmids.Methods The complete clinical data of fresh tumour tissue and adjacent non-tumour tissue wax blocks of 111 gastric cancer patients who underwent radical surgery in the Cancer Hospital of Tianjin Medical University from January 2004 to September 2007 were collected.Immunohistochemical method was used to detect the expression level of RNF212 in cancer and adjacent tissues.Semi-quantitative reverse transcription and polymerase chain reaction(Sq RT-PCR)and Western Blot were used to detect the m RNA and protein expression levels of RNF212 in 7 gastric cancer cell lines and a normal gastric mucosa cell line.The gastric cancer cell line MGC-803 was selected and transfected with RNF212 interfering plasmid.The effect of oncogene RNF212 on gastric cancer cell proliferation was studied through CCK8(Cell Counting Kit-8)and cloning formation experiments.Results 1.Immunohistochemical results showed that the positive expression rate of RNF212 in gastric cancer tissue was 66.67%(74/111)and the negative expression rate was 33.33%(37/111),the former was significantly higher than the latter,and the difference was statistically significant Significance(P = 0.008).In the study of the relationship between the expression level of RNF212 and the clinicopathological characteristics and prognosis of gastric cancer patients,it was found that the age and p N stage of the patients were correlated with the high expression of RNF212(P = 0.021;P = 0.002).2.When comparing the RNF212 interference group cells with the control group cells through CCK8 and clone formation experiments,it was found that the number of clones in the interference group cells was significantly reduced,and the migration and invasion ability of the interference group cells in the Transwell experiment was significantly weaker than that of the control group.Conclusions 1.The abnormal expression of RNF212 gene is related to the occurrence and development of gastric cancer lesions,and has important guiding significance for early screening of gastric cancer.2.The expression of RNF212 in gastric cancer tissues is significantly higher than that of adjacent non-tumour tissues.The high expression of RNF212 is significantly associated with positive lymphatic infiltration and the age of surgery.The high expression of RNF212 is associated with a low survival rate,and has been identified as an independent prognostic factor for GC in multivariate analysis.The high expression of RNF212 is an independent prognostic factor in patients with gastric cancer,and its expression level in gastric cancer tissues can be used to assess the patient’s risk and guide its treatment.3.RNF212 can promote the proliferation and migration of cells,thereby promoting the progress of gastric cancer. |
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