The Expression Of Oncogene RMP In Non-small Cell Lung Cancer(NSCLC) And Its Effect On Carcinoma’s Biological Property

Objective:In our research, we study the relationship between RMP and the clinical pathological features of NSCLC patients by measuring the expression level of RMP in human non-small lung cancer tissues and cell lines. At the same time, we explore the influence of RMP on carcinoma’s biological functions to provide powerful supports for gene targeting treatment of NSCLC.Methods:1. q RT-PCR and western blot were used to detect the m RNA and protein expression in NSCLC cell lines and the human normal bronchial epithelial cells.2. q RT-PCR and western blot were used to detect the m RNA and protein expression in NSCLC tissues and adjacent non-tumor tissues.3. q RT-PCR was applied to detect the expression of RMP in NSCLC tissues which have different characteristics. Then we analyzed the relationship between RMP and the tumors. At the same time, we used the table to summarize the relevant clinical information about the patients.4. Fluorescence microscope was used to observe the different RMP plasmids in A549 cells which belong to NSCLC. And we also successfully detected the transfection efficiency by using the q RT-PCR and western blot.5. CCK-8 assay was applied to the influence of RMP on cell proliferation of A549.6. A549 cells transfected with different groups of RMP plasmids were dealt with Gefitinib and then we used the CCK-8 assay and Flow cytometry to detect the effect of RMP on the sensitivity of A549 cells to chemotherapy.7. CCK-8 assay and Flow cytometry were applied to observe the effect of RMP on the sensitivity of A549 cells to radiotherapy. At the same time, we used western blot to detect the protein expression of apoptosis related genes.8. Flow cytometry was used to detect the effect of RMP on the cell cycle of A549 cells. At the same time, western blot was applied to observe the protein expression of cell cycle related genes.9. The migration and invasion assay was used to detect the effect of RMP on the ability of migration and invasion in A549 cells.10. The xenografts in nude mices were used to observe the influence of RMP on tumor formation and we also applied the immunohistochemical staining to detect the protein expression of RMP、Bax、Bcl2 and capase-3.Results1. q RT-PCR and western blot showed that both m RNA and protein level of RMP were relatively high expressed in NSCLC cells.2. q RT-PCR and western blot showed that both m RNA and protein level of RMP were distinctly high expressed in human non-small cell lung cancer tissues compared with corresponding normal tissues. At the same time, we found that the expression of RMP was associated with the status of lymphonodus and T-stage in carcinoma tissues. In addition, there was nothing to do with the patients’ age、gender and cancer forms.3. Fluorescence microscope、q RT-PCR and western blot showed that each group of RMP plasmids were transfected into A549 cells successfully. As a result, the m RNA and protein level were relatively improved.4. CCK-8 assay showed that RMP could distinctly promote the proliferation of A549 cells.5. CCK-8 assay showed that RMP could still slow down the death induced by different dose of chemotherapy agents. Apart from that, the Flow cytometry showed that RMP could decrease the apoptosis induced by chemotherapy.6. CCK-8 assay indicated that RMP could still inhibit the death induced by different dose of radiation. Additionally, the Flow cytometry showed that RMP could decrease the apoptosis induced by radiotherapy in A549 cells. What’s more, western blot demonstrated that RMP could suppress the protein expression of apoptosis related genes while improve that of anti-apoptosis related genes after the radiotherapy.7. The Flow cytometry showed that RMP could decrease the G2 phase arrest induced by the radiotherapy in A549 cells. What’s more, western blot demonstrated that RMP could regulate the protein expression of cell cycle related gene after the radiotherapy.8. The migration and invasion assay showed that RMP could improve the ability of migration and invasion in A549 cells.9. The xenografts in nude mices indicated that RMP could promote the growth of xenografts. At the same time, immunohistochemical staining showed that RMP could improve the tumor growth by restraining the protein expression of apoptosis related genes while promoting that of anti-apoptosis related genes.Conclusions1. RMP was highly expressed in non-small cell lung cancer. In addition, we found that the RMP expression was associated with the status of lymphonodus and T-stage by combining the patients’ relevant clinical information. All these indicated that RMP may be closely related to the growth and malignant degree of NSCLC. Furthermore, RMP would be a potential NSCLC related cancer-promoting gene.2. Overexpression of RMP could change a variety of biological functions such as cell proliferation、apoptosis、cell cycle、migration and invasion in A549 cells leading to promote the growth of xenografts in nude mices. All these further showed that RMP would play a cancer-promoting gene role in NSCLC which may provide us a powerful goal for gene targeting treatment of lung cancer.