KiSS-1基因对滋养细胞生物学行为影响的实验研究

目的 :探讨KiSS 1基因对滋养细胞生物学行为的直接影响 ,进一步证实妊娠高血压综合征患者胎盘“浅着床”与滋养细胞中肿瘤转移抑制基因KiSS 1表达增高有关。方法 :含有人KiSS 1基因全长cDNA的重组真核表达载体pcDNA3 KiSS 1经酶切和测序鉴定正确后 ,转染滋养细胞株JAR ,观察其KiSS 1mRNA表达、细胞生长和增殖能力和细胞侵袭力的变化。结果 :pcDNA3 KiSS 1成功转染JAR细胞 ,KiSS 1mRNA表达阳性 ,而亲本JAR细胞为阴性。转染目的基因后细胞生长曲线、软琼脂克隆形成率等与未转染之细胞无差异 ,但细胞侵袭能力明显下降 ,穿透Matrigel膜细胞数明显少于未转染组。结论 :KiSS 1基因对滋养细胞的生长、增殖能力无影响 ,但抑制其侵袭能力 ,这可能是导致妊高征“浅着床”的原因

The experimental study:effect of KiSS-1 gene on the biological behaviors of trophoblast cell

Objective:To investigate the effect of KiSS-1 gene on the biological behaviors of trophoblast cell in order to explore the relationship between shallow implantation of placenta and high expression of KiSS-1 gene of placental villi in pregnancy induced hypertension syndrome.Methods:The recombinant eukaryotic expression vector (pcDNA3-KiSS-1) inserted by the full-length cDNA encoding human KiSS-1 gene was constructed and confirmed by enzyme digest identification and DNA sequencing.The pcDNA3-KiSS-1 vector was transfected into human trophoblast cell line JAR by lipofectamine.The characteristics in the KiSS-1 transfected cells include expression of KiSS-1mRNA,ability of growth,proliferation and invasion were observed.Results:KiSS-1 gene was integrated into the genomic DNA of JAR.Expression of KiSS-1mRNA was positive in transfected cell and negative in untransfected cell.The comparison of cell proliferation curves and clone formation rate between the transfected and untransfected group had no significant different.Boyden chamber invasion assay show the cell numbers invading through matrigel filter was significantly decreased in KiSS-1 transfected group compared with untransfected group.Conclusion:KiSS-1 gene suppresses invasion of trophoblast cell in vitro without affecting growth and proliferation.It may play a major role in shallow implantation of placenta in pregnancy induced hypertension syndrome.