The Expression Of Oncogene RMP In Esophageal Squamous Cell Carcinoma (ESCC) And Its Correlation With Tumor Invasion And Metastasis And Epithelial-mesenchymal Transition

Objective:The aim of this research is to investigate the expression level of oncogene RMP in human esophageal squamous cell carcinoma tissues and cell lines and to explore its correlation with tumor invasion and metastasis and Epithelial-Mesenchymal Transition(EMT). Methods:1. The expression of RMP mRNA and protein in ESCC tissues and adjacent non-tumor tissues was detected by qRT-PCR and Western blot. Then we analyzed the correlation with RMP and clinicopathologic features.2. The expression of RMP mRNA and protein in ESCC cell lines and the human normal esophageal epithelial cells was detected by qRT-PCR and Western blot.3. qRT-PCR was applied to detect the different RMP plasmids mRNA expression in Eca109 cells which belong to ESCC and CCK-8 assay was used to measure the transient transfected Eca109 cells proliferation viability.4. Cell adherence assay was applied to detect the effect of RMP on the adhering capacity of Eca109 cells.5. Wound healing assay was used to measure the effect of RMP on the migration ability of Eca109 cells.6. Transwell assay was applied to observe the effect of RMP on the migration and invasion of Eca109 cells.7. Western blot was used to detect the protein expression of E-cadherin, N-cadherin and Vimentin in Eca109 cells. Results:1. qRT-PCR and Western blot indicated that both RMP mRNA and protein expression level in human ESCC tissues were higher than corresponding adjacent tissues. Meanwhile, we found that RMP expression level was correlated with and T-stage and the status of lymphonodus. However, Age, gender and tumor location were not associated with the expression of RMP.2. qRT-PCR and Western blot indicated that both RMP mRNA and protein expression level in ESCC cell lines were higher than the human normal esophageal epithelial cells.3. qRT-PCR indicated that the different RMP plasmids were transfected into Eca109 cells successfully. The RMP mRNA expression level was relatively improved.4. CCK-8 assay indicated that RMP could promote the proliferation of Eca109 cells.5. Cell adherence assay showed that RMP could reduce cell adhesion ability in Eca109 cells.6. Wound healing assay showed that RMP could promote cell migration in Eca109 cells.7. Transwell assay showed that RMP could improve cell migration and invasion ability in Eca109 cells.8. Western blot demonstrated that RMP overexpression could reduce the E-cadherin protein expression and promote the N-cadherin and Vimentin protein expression. However, depletion of RMP expression will not affect all of them. Conclusions:1. RMP was highly expressed in human ESCC tissues and ESCC cell lines and combined the correlation with RMP and clinicopathologic features indicated that RMP may play an important role in the development of ESCC. All these results showed that RMP would be a potential ESCC related cancer gene.2. RMP overexpression could improve cell metastasis and invasion ability in vitro. However, RMP depletion could restrain this phenomenon. RMP could promote EMT in ESCC cells by downregulating the E-cadherin protein expression and upregulating the N-cadherin and Vimentin protein expression.