5-8F Nasopharyngeal Carcinoma Cell Line And LEC Morphological Study Of The Interaction Of Cell Lines

Objective:For statistical observation of nasopharyngeal carcinoma cell line 5-8F cell lines and H-LEC morphological changes after the interaction,to lay the foundation for lymph node metastasis and related research to further study the mechanism of NPC.Methods:Selected different MOI values transfect Nasopharyngeal carcinoma line 5-8F cells with red fluorescence-labeled lentivirus as transfection vector.RFP-5-8F cells were cultured and then were purified after transfection.Transfection efficiency was determined by flow cytometry after transfection.RFP-5-8F cells transfected optimal MOI values was considered as the experimental group,and the untransfected parental 5-8F cells for the control group.MTT and Wound healing assays were used to detect the migration and proliferation of 5-8F and RFP-5-8F in logarithmic phase,and the morphology of these cells were observed under microscope.The cell growth curve and migration ability was analyzed.Logarithmic growth phase of the RFP-5-8F,GFP-LEC cells were co-cultured cell density suitable for live cell observation stations.Logarithmic growth phase of the RFP-5-8F,GFP-LEC cells,using easy to wound healing experiments will Di dedicated plug-line scratch test,repair capacity was observed to migrate under the co-culture cells.Take RFP-5-8F cells cultured alone were cultured RFP-5-8F + GFP-LEC cells were seeded in BALB/c nude mice armpit observed tumor formation in nude mice,in vivo fluorescence imaging,lymph node metastasis in nude mice.BD Matrigel in Matrigel shop at the bottom of the dish,the take individual GFP-LEC tubule cells after forming ability and RFP-5-8F + GFP-LEC co-cultured lymphatic endothelial cells was observed.Application SPSS 16.0 statistical analysis software to analyze data,P<0.05 was considered statistically significant.Results:Transfection efficiency of 5-8F cells were higher than 95%by flow cytometry,and the best MOI was 30 with moderate endoscopic fluorescence intensity.Compared with the control group,5-8F and RPP-5-8F cells in the experimental group were morphologically similar under microscope,and the growth curve of both were in consistent,without statistical significance(P=0.997).Wound healing assays showed a consistent 5-8F and RFP-5-8F cell migration ability,without statistical significance(P>0.05).Workstation observe living cells co-cultured RPP-5-8F,GFP-LEC cells induce GFP-LEC around tumor cells(RFP-5-8F)line into the tube-like structure.RFP-5-8F co-cultured with GFP-LEC cell migration capacity greater than a single GFP-LEC cultured cells,the difference was statistically significant(P<0.05).And GFP-LEC co-cultured RFP-5-8F cells cultured alone compared with RPP-5-8F has more lymph node metastasis.Tubule-forming ability and RFP-5-8F co-cultured with GFP-LEC tube formation ability cultured alone was no significant difference,the difference was not statistically significant(P>0.05).After the RFP-5-8F cells with GFP-LEC local co-cultured tumor growth in nude mice compared with separate RFP-5-8F of slowed down,but lymph node metastasis occurs earlier and more pronounced.Conclusion:Lentivirus red fluorescent protein had no significant effect on the biological characteristics of 5-8F cells,RFP-5-8F 5-8F behalf for subsequent experiments.RFP-5-8F,after GFP-LEC cells were co-cultured with GFP-LEC promote the ability to enhance the RFP-5-8F migration and the formation of tube-like structure similar;enhanced tumor cell migration RFP-5-8F;Nasopharyngeal carcinoma cells in contact with the lymphatic endothelial cells and interacts may be an important reason for nasopharyngeal carcinoma lymph node metastasis,the mechanism needs further study path.