The Effect Of Sigma-1 Receptor On The Biological Function Of Trophoblast Cell Line HTR-8/SVneo Cells

Background and ObjectivePregnancy is a multi-link process,and any link is an important factor that affects the success of pregnancy.The placenta acts as a barrier between the mother and the fetus,providing the fetus with the nutrients it needs to develop.At present,more and more studies begin to focus on the relationship between the placenta and abnormal pregnancy.Trophoblast cells are an important part of placental tissue in early pregnancy and play an important role in the development of placenta and fetus.Proliferation,migration and invasion of trophoblast cells play a crucial role in the successful establishment and maintenance of pregnancy.There is a certain relationship between the impaired function of trophoblast cells and the occurrence of abortion,but the underlying mechanism of its occurrence is not very clear at present.The study of the molecular regulation mechanism of trophoblast cells can provide a new direction for the prevention and treatment of spontaneous abortion.The Sigma-1 receptor is a ligand-operated protein,mainly located in the mitochondria-associated endoplasmic reticulum membrane.The sigma-1 receptor was originally thought to be an opioid receptor,but is now considered to be an unusual molecular chaperone.Studies have shown that the sigma-1 receptor is involved in the development of a wide range of diseases,not only neurodegenerative diseases,but also heart disease,diminished ovarian reserve,cancer and so on.Sigma-1 receptor is highly expressed in human extravillous trophoblast cells,and the study of sigma-1 receptor in extravillous trophoblast cells has not been conducted yet.We hypothesized that the Sigma-1 receptor may induce abortion by affecting the biological behavior of trophoblast cells.In this study,we will investigate the effects of Sigma-1 receptor antagonist BD1063 and small-interfering ribonucleic acid(siRNA)targeting Sigma-1 receptor on the biological behaviors of proliferation,migration and invasion of trophoblast cell line HTR-8/SVneo cells to provide new ideas for the prevention and treatment of abortion.Materials and methodsHTR-8/SVneo cells were treated with sigma-1 receptor antagonist BD1063 and siRNA targeting Sigma-1 receptor.The expression level of sigma-1 receptor was detected by q RT-PCR and Western blotting.MTT assay was used to detect the effect of sigma-1 receptor antagonist BD1063 and siRNA targeting Sigma-1 receptor on the proliferation of HTR-8/SVneo cells.The effect of sigma-1 receptor antagonist BD1063 and siRNA targeting Sigma-1 receptor on the migration ability of HTR-8/SVneo cells was detected by wound healing assay.Transwell assay was used to detect the effect of sigma-1 receptor antagonist BD1063 and siRNA targeting Sigma-1 receptor on the invasion ability of HTR-8/SVneo cells.Results1.Immunohistochemical results of villi tissue showed: the expression of sigma-1receptor in villus tissue of normal early pregnancy induced abortion patients was significantly higher than that of early spontaneous abortion patients(P < 0.01);2.Effect of siRNA targeting Sigma-1receptor transfection and sigma-1 receptor antagonist BD1063 on sigma-1 receptor expression levels: After siRNA targeting Sigma-1receptor transfected HTR-8/SVneo cells,the m RNA expression of sigma-1receptor was significantly reduced(P < 0.0001),and the protein expression level of sigma-1 receptor was also significantly reduced(P < 0.01),the protein expression level of sigma-1 receptor was significantly decreased after the treatment of sigma-1receptor antagonist BD1063(P < 0.01);3.Effect of Sigma-1 receptor on proliferation ability of trophoblast cells: After treatment with Sigma-1 receptor antagonist BD1063 for 24 h,the proliferation of HTR-8/SVneo cells was significantly inhibited with the increase of BD1063 concentration.The IC50 value of BD1063 was 7μmol/L.After 48 h treatment with siRNA targeting Sigma-1 receptor,the proliferation of HTR-8/SVneo cells was also significantly inhibited compared with the negative control group(P < 0.05);4.Effect of Sigma-1 receptor on the migration of trophoblast cells: After treated with 7μmol/L Sigma-1 receptor antagonist BD1063 for 24 h,the migration of HTR-8/SVneo cells was significantly inhibited compared with the control group(P <0.01).After treated with siRNA targeting Sigma-1 receptor for 48 h,the migration ability of HTR-8/SVneo cells was also significantly inhibited compared with the negative control group(P < 0.01);5.Effect of Sigma-1 receptor on invasion ability of trophoblast cells: After treated with 7μmol/L Sigma-1 receptor antagonist BD1063 for 24 h,the invasion ability of HTR-8/SVneo cells was significantly inhibited compared with control group(P <0.01).After 48 hours of siRNA targeting Sigma-1 receptor treatment,the invasion ability of HTR-8/SVneo cells was also significantly inhibited compared with the negative control group(P < 0.01).Conclusions1.The expression of sigma-1 receptor in villus tissue of normal early pregnancy induced abortion patients is significantly higher than that of early spontaneous abortion patients;2.Treatment of HTR-8/SVneo cells with Sigma-1 receptor antagonist BD1063 and siRNA targeting Sigma-1 receptor can reduce the expression of sigma-1receptor;3.Treating HTR-8/SVneo cells with sigma-1 receptor antagonist BD1063 and siRNA targeting Sigma-1 receptor can reduce the proliferation,migration and invasion of trophoblast cells,suggesting that sigma-1 receptor is involved in the regulation of trophoblast cell biological behavior.