| Objective: The incidence of nasopharyngeal carcinoma(NPC)in Guangdong and Guangxi provinces is high in China.NPC has a high degree of concealment,malignancy and metastasis,which seriously threatens the patient's health.In recent years,a great number of studies have found that inactivation of tumor suppressor genes is an important factor in the occurrence and development of NPC.It is found that stromal cell derived factor 2 like 1(SDF2L1)downregulation is relationship with the occurrence and prognosis of tumor.This study aimed to investigate the expression of SDF2L1 in NPC tissue and its effect on the biological function of NPC cell.Methods: 1.The expression of SDF2L1 mRNA and protein in NPC tissues and chronic inflammation of nasopharyngeal mucosa tissues was detected by quantitative real-time polymerase chain reaction(qRT-PCR)and immunohistochemistry(IHC),and analyzed its relationship with clinical pathological features.2.The expression of SDF2L1 mRNA and protein inNP69,CNE1 and CNE2 Z cells was detected by qRT-PCR and western blotting(WB).3.CNE1 cells were transfected by lentiviral vector containing RNA interference fragments and selected by puromycin.SDF2L1 RNA interference fragments in CNE1 cells were defined as experimental group.Negative RNA interference fragments in CNE1 cells were defined as empty control group.Untransfected CNE1 cells were defined as control group.WB was used to evaluate the expression of SDF2L1 in different groups.4.CNE1 cell proliferation was by MTT assays and colony formation assays.5.The migration and invasion abilities of silencing SDF2L1 in CNE1 cells were detected by wound-healing,transwell migration and transwell invasion.6.The relative mRNA expression of E-cadherin,N-cadherin and matix metalloproteinase-9(MMP-9)was detected by qRT-PCR.Results: 1.The relative expression of SDF2L1 mRNA in 12 cases of NPC tissues and 12 cases of chronic inflammation of nasopharyngeal mucosa tissues was 0.549?0.568 and 1.254?0.729,respectively.The relative expression of SDF2L1 mRNA in NPC tissues was significantly lower than that in chronic inflammation of nasopharyngeal mucosa tissues,and the difference had statistical significance((49)(27)0.05).The positive rate of SDF2L1 protein expression in NPC tissues was 20.6%(20/97),which was significantly lower than its positive rate of 91.4%(53/58)in chronic inflammation of nasopharyngeal mucosa tissues.The difference was statistically significant((49)(27)0.05).The expression of SDF2L1 protein in NPC tissues was not obvious associated with clinical pathological parameters((49)(29)0.05).2.The relative expression of SDF2L1 mRNA in NP69 cells,CNE1 cells and CNE2 Z cells was 1.006 ? 0.137,0.638?0.048 and 0.287?0.016,respectively.The relative expression of SDF2L1 protein in NP69 cells,CNE1 cells and CNE2 Z cells was 0.980 ? 0.019,0.932?0.025 and 0.517?0.036,respectively.The relative expression of SDF2L1 mRNA and protein in CNE1 and CNE2 Z cells was significantly lower than that of NP69 cells,and the difference was statistical significance((49)(27)0.05).3.The relative expression of SDF2L1 protein in the experimental group,the empty vector group and the control group was 0.280 ? 0.039,0.935 ? 0.034 and 0.926? 0.031,respectively.The relative expression of SDF2L1 protein in the experimental group was obviously lower than that of the empty vector group and the control group,and the difference had statistically significant((49)(27)0.05).4.In MTT assays,the OD values of 96 h in the experimental group,the empty vector group and the control group were 1.135?0.013,1.149?0.021 and 1.127?0.019,respectively.There was no significant difference in cell proliferation ability in each group,and the results were not statistically significant((49)(29)0.05).In colony formation assays,the rate of cell colony formation in the experimental group,the empty vector group and the control group was(60.4 ? 6.39)%,(58.67 ? 5.75)% and(59.41 ? 5.02)%,respectively.There was no significant difference in cell colony formation in each group,and the results were not statistical significance((49)(29)0.05).5.In wound-healing assays,the rate of relative cell migration in the experimental group,the empty vector group and the control group was(72.79 ? 0.96)%,(53.06 ? 0.74)% and(52.38 ? 0.65)%,respectively.The rate of relative cell migration in the experimental group was obviously higher than that of the empty vector group and the control group,and the difference was statistically significant((49)(27)0.05).In transwell migration assays,the number of cells penetrated into the lower chamber in the experimental group,the empty vector group and the control group was 331?7,254?13 and 242?9,respectively.The number of cells penetrated into the lower chamber in the experimental group was significantly higher than that of theempty vector group and the control group,and the difference had statistical significance((49)(27)0.05).In transwell invasion assays,the number of cells penetrated into the lower chamber in the experimental group,the empty vector group and the control group was 221?10,140?9 and 146?7,respectively.The number of cells penetrated into the lower chamber in the experimental group was obviously higher than that of the empty vector group and the control group,and the results were statistically significant((49)(27)0.05).6.Comparison with the empty vector group and the control group,in the experimental group,the relative expression of E-cadherin mRNA was significantly decreased((49)(27)0.05)and the relative expression of MMP-9 mRNA was obviously increased((49)(27)0.05),and the relative expression of N-cadherin mRNA was not obviously difference((49)(29)0.05).Conclusions: 1.The expression of SDF2L1 mRNA and protein in NPC tissues and NPC cells was down-regulation,and its expression was not obviously associated with clinical pathological characteristics.2.The low expression of SDF2L1 in NPC CNE1 cells has been successfully established.3.SDF2L1 silencing promoted the migration and invasion of NPC cells,but it was not significant effect on the proliferation of NPC cells.4.The low expression of SDF2L1 in NPC cell line resulted in the expression changes of EMT related E-cadherin and MMP-9,indicating that down-regulation of SDF2L1 may be a molecular marker for invasion and distant metastasis of NPC and may be involved in the occurrence and development of NPC. |
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