| Objectives To investigate the biological behavior effect on human gastric cancer cell line AGS,after siRNA silent NRP-2 of it.It provided a theoretical basis that NRP-2 could be a new target for the treatment of gastric cancer.Methods Strict standard culture of human gastric cancer cell line AGS;Using liposome Lip-2000 as carrier,siRNA with fluorescent NRP-2 was transfected into the cell line,After 48 hours of transfection,the transfection efficiency was detected under the fluorescence microscope,and the transfection efficiency can be considered to be more than 60 %,and the transfection proportion is recorded.The cells were divided into three groups: In transfection group,siRNA with silencing NRP-2 was transfected into AGS cell line.blank control group: the PBS was transfected into the cell line;no significant sequence group: the control-siRNA of NRP-2 was transfected into gastric cancer cell line cell strain;Using quantitative real-time PCR(qPCR)expression level differences in the three groups of NRP-2 to detect the level of mRNA;MTT assay was used to detect the proliferation of three groups of cell lines;The expression of protein Ki-67 in three groups of cell lines was detected by immunocytochemical staining;Flow cytometry was used to detect the apoptosis of the three progenitor cells;Flow cytometry was used to detect the apoptosis of the cells;Transwell small cell experiment was used to detect the invasion and migration of the three-progenitor cell line,and the statistical data was used to analyze the difference and the conclusion was drawn.Results 1 Fluorescence microscopy results: Liposome mediated Lip-2000 fluorescent NRP-2-siRNA was successfully transfected into gastric cancer cell line AGS,and the transfection efficiency was more than 80%;2 The results of qPCR: The relative expression of NRP-2 mRNA in the transfected group of gastric cancer AGS cells was significantly lower than that in the blank control group and the non significant sequence group.The difference was statistically significant(P < 0.05).3 The results of MTT were as follows: The proliferation ability of AGS cells in the transfected group was significantly lower than that of the control group and the non significant sequence group.The difference was statistically significant(P < 0.05).4 The results of Immunocytochemistry staining: The relative expression of Ki-67 protein in the transfected group of gastric cancer AGS cell lines was significantly lower than that in the blank control group and the non significant sequence group.The difference was statistically significant(P < 0.05).5 The results of Flow cytometry: In the transfected group,the gastric cancer AGS cells stagnated at Gl stage were significantly more than that of the blank control group and the non significant sequence group.These results indicate that the growth cycle of gastric cancer cells in transfection group is inhibited.The difference was statistically significant(P < 0.05).6 The results of Transwel chamber migration experiment: the migration capacity of the gastric cancer cells of the transfected group is obviously lower than that of the blank control group and the meaningless sequence group.The difference was statistically significant(P < 0.05).7 The results of Transwel chamber invasion experiment: The invasiveness of AGS cells in the transfected group was significantly lower than that in the blank control group and the non significant sequence group.The difference was statistically significant(P < 0.05).Conclusions Transfection of silencing NRP-2 siRNA into human gastric cancer AGS cells by liposome Lip-2000 resulted in a series of changes in biological behavior: The relative expression of NRP-2-mRNA is reduced;The cell proliferation ability decreased significantly;The relative expression of Ki-67 protein decreased significantly;The cell growth cycle is obviously inhibited;Cell migration,invasion ability decreased significantly.It provides a theoretical basis for NRP-2 to be a new target for the treatment of gastric cancer. |
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