Effects Of Different Concentrations Of Prolactin On Proliferation,Migration And Invasion Of Trophoblast Cells In Human Early Pregnancy

Objective:To investigate the effects of different concentrations of prolactin(PRL)on the proliferation,migration and invasion of human trophoblast cell line HTR-8 \ Snveo.Methods:HTR-8\Snveo cells were used as the study group.The experiment was divided into 3 groups: blank group(0ng/mL),low concentration PRL(10ng/mL)stimulation group,high concentration PRL(100ng/mL)stimulation group.2.To investigate whether PRL has an effect on migration and invasion of HTR-8\Snveo cells by using cell scratches repair experiments and transwell chamber invasion experiments.The CCK-8 colorimetric assay was used to detect the proliferation of experimental cells in 24 h after intervention with different concentrations of PRL.The expression and secretion of MMP-2 and E-cadherin were detected by real-time fluorescence quantitative PCR(RT-PCR)and double antibody sandwich ELISA respectively.Result:The results of cell scratch repair showed that the migration rate(%)of the experimental cells was 28.02±0.75,37.80±0.42,42.12±0.31,respectively.The difference between the groups was statistically significant(P < 0.05).Transwell invasion assay results showed: compared with the control group(100±6.56),the HTR-8/SVneo cell invasion index of 10ng/mL and 100ng/mL concentration of PRL after 24 h incubation(142.68±4.91,189.59±4.10,P<0.05)increased significantly,and HTR-8/SVneo cells increased with the concentration of PRL invasion index also increased,the difference was statistically significant.The CCK-8 method was used to detect the cell proliferation ability.The results showed that the OD value of the control group was 1.08±0.04,the experimental group was 1.09±0.05,1.13±0.06,P > 0.05,the difference was not statistically significant.RT-PCR and Elisa experimental data analysis results show that,compared with the control group,different concentration of PRL were significantly up-regulated the mRNA and protein expression level of MMP-2(P< 0.05),downregulation of mRNA E-cad and protein expression level(P <0.05),and showed a concentration dependent effect.Conclusion: 1.In the experimental concentration range,PRL can promote the migration and invasion ability of HTR-8/SVneo cells in vitro,and with the increase of PRL concentration,the migration and invasion ability increased.2Different concentrations of PRL(0,10,100ng/mL)had no effect on the proliferation of HTR-8\SVneo cells.3 PRL in the range of the experimental concentration can increase the expression of MMP-2 and protein in mRNA,and down regulate the expression of E-cad and protein in the mRNA,which can promote the migration and invasion of HTR-8/SVneo cells.