hCG-β反义寡脱氧核苷酸对人绒癌JAR细胞hCG分泌作用的研究

Effects of Antisense Oligodeoxynucleotides of hCG-β on Production of hCG by JAR Cells in Vitro

目的 :探讨人绒毛膜促性腺激素 -β反义寡脱氧核苷酸 (h CG-β AS-ODN)抑制 JAR绒癌细胞株 h CG的分泌。方法 :设计 h CG-β m RNA的 AS-ODN,作用于体外培养的 JAR绒癌细胞株 ,细胞浓度为 1× 1 0 5/个。ODN的浓度分别为 5 μmol/ L、1 0 μmol/ L、2 0 μmol/ L 与 JAR细胞共培养在96孔培养板中 ,2 4h、48h、72 h和 96 h,取上清测 h CG-β含量。结果 :1 0μmol/ L浓度的 h CG-βAS-ODN对 JAR细胞在体外作用 2 4h,h CG-β分泌呈下降趋势 ,48h下降达 5 7% ,此后 ,抑制作用略有减弱 ,而 2 0 μmol/ L的正义和随机的 ODN,分泌也受抑制。结论 :5～ 1 0 μmol/ L 的 AS-ODN明显抑制体外培养的 h CG分泌细胞 h CG蛋白的分泌 ,且该 AS-ODN有希望作为 h CG分泌抑制物应用。

Objective:To observe the inhibition effect of antisense oligodeoxynucleotides(AS ODN) of hCG β on hCG secretion by JAR cells.Methods:A AS ODN was synthesized in accordance with the mRNA of the hCG β subunit.The human chorionic carcinoma cell line JAR was used for research.The terminal concentration of the cell suspension was 1×10 5 cell/ml.The suspension for each well containing 5 μmol/L,10 μmol/L and 20 μmol/L antisense respectively were added to 96 well microtitre culture plate,cultured for 24 h,48 h,72 h and 96 h.The supernatants were collected for detecting hCG β.Results:The concentration of 10 μmol/L AS ODN of hCG β caused a significant inhibition of secreting hCG by JAR cells in 24 h culture in vitro.The inhibition rate reached to 57% and began to decline after 48 h.When the concentration reached to 20 μmol/L,both sense and random ODN showed a significant inhibition on secreten of hCG β.Conclusion:The concentrations of 5～10 μmol/L AS ODN of hCG β caused significant inhibition of HCG production of hCG secretion cell cultured in vitro.the AS ODN of hCG β maybe used as a inhibitor of hCG secretion.