子痫前期滋养细胞ABCG2表达的研究

目的探讨正常及子痫前期胎盘滋养细胞分化前、后胎盘表达的主动转运因子ATP结合匣式转运子G2 (ABCG2)mRNA及蛋白的表达变化。方法酶消化法原代培养正常及子痫前期胎盘绒毛滋养层细胞,经传代与纯化后免疫鉴定滋养细胞及其纯度。收集培养后72 h (未分化状态)及培养后6 d (分化状态)的细胞,裂解后应用实时荧光定量PCR及Western Blot检测各组细胞ABCG2 mRNA及蛋白的表达水平。结果成功自正常及子痫前期胎盘中原代培养人绒毛滋养层细胞,未分化状态下子痫前期滋养细胞ABCG2mRNA及蛋白的表达平均值为正常的2. 54及2. 06倍,分化状态下子痫前期滋养细胞ABCG2mRNA及蛋白的表达为正常的1. 65及1. 53倍;正常胎盘滋养细胞分化后ABCG2 mRNA及蛋白表达增高3. 55及4. 06倍,子痫前期组仅增加2. 31及3. 03倍。结论自子痫前期及正常胎盘原代培养滋养细胞区分不同分化状态,实时荧光定量PCR及Western Blot检测子痫前期滋养细胞不同分化时期ABCG2 mRNA及蛋白的表达均高于正常,但滋养细胞分化后子痫前期ABCG2 mRNA及蛋白表达增长的程度低于正常。

Study on ABCG2 expression in trophoblast cells of preeclampsia patients

Objective To explore the changes of ABCG2 mRNA and protein expressions before and after the differentiation of primary cultured trophoblast cells from normal and preeclamptic placentas. Methods Human villous trophoblast cells from normal and preeclamptic placentas were cultivated by enzyme digestion,after cell passage and purification,the purity of trophoblast cells was identified by immunoassay,the expression of ABCG2 mRNA and protein in trophoblast cells cultivated 72 hours( before differentiation) and 6 days( after differentiation) were detected by real-time fluorescence quantitative PCR and Western Blot. Results Human villous trophoblast cells of normal and preeclamptic placentas were cultivated successfully. The mean values of ABCG2 mRNA and protein expressions in trophoblast cells before differentiation were 1. 54 and 1. 06 times more than the normal trophoblast cells,respectively;the mean values of ABCG2 mRNA and protein expressions in trophoblast cells after differentiation were 0. 65 and 0. 53 times more than the normal trophoblast cells,respectively. After differentiation,the expressions of ABCG2 mRNA and protein in normal trophoblast cells increased by 3. 55 and 4. 06 times,respectively,the expressions of ABCG2 mRNA and protein in preeclamptic trophoblast cells increased by 2. 31 and 3. 03 times, respectively.Conclusion The expressions of ABCG2 mRNA and protein in different differentiation stages of preeclampsia are higher than normal placentas detected by real-time quantitative PCR and Western Blot. However,the elevation degrees of ABCG2 mRNA and protein expressions in preeclampsia are lower than normal placentas after differentiation of trophoblast cells.