体外培养的人胎盘细胞滋养层细胞中钠钾ATP酶的活性与表达

Activity and expression of Na(+)-K(+)-ATPase in human placental cytotrophoblast cells in culture.

Clarson L H, Glazier J D, Greenwood S L, Jones C J, Sides M K, Sibley C P

Department of Child Health and School of Biological Sciences, University of Manchester, St Mary's Hospital, UK.

J Physiol. 1996 Dec 15;497 ( Pt 3)(Pt 3):735-43. doi: 10.1113/jphysiol.1996.sp021804.

Abstract

To determine whether there is a change during differentiation, the activity and expression of Na(+)-K(+)-ATPase were studied in mononucleate cytotrophoblast cells (18 h culture) and syncytiotrophoblast-like cells (66 h culture). A choriocarcinoma-derived cell line (JAr) which, unlike the cytotrophoblast cells, divides in culture, was also studied for comparison. 2. Na(+)-K(+)-ATPase activity was assessed by measurement of ouabain-sensitive 86Rb+ uptake. Na(+)-K(+)-ATPase expression was determined by (i) measurement of [3H]ouabain binding and (ii) Northern hybridization to measure expression of alpha-1 and beta 1-subunit mRNA. 3. There was no significant difference in either activity or expression of Na(+)-K(+)-ATPase during differentiation of cytotrophoblast cells. However, expression of alpha 1- and beta 1-subunit mRNA was significantly lower in 66 vs. 18 h cultured cytotrophoblast cells. 4. Both Na(+)-K(+)-ATPase activity and [3H]ouabain binding was significantly greater in JAr cells than either cytotrophoblast cell groups, although expression of alpha 1- and beta 1-subunit mRNA was the same as cytotrophoblast cells cultured for 18 h. 5. It is concluded that N(+)-K(+)-ATPase activity and protein expression does not change during differentiation of cytotrophoblast cells but that there are changes in expression at the transcriptional or post-transcriptional level.

摘要

为了确定分化过程中是否存在变化，我们研究了单核细胞滋养层细胞（培养18小时）和类合体滋养层细胞（培养66小时）中Na(+)-K(+)-ATP酶的活性和表达。为作比较，还研究了一种源自绒毛膜癌的细胞系（JAr），该细胞系与细胞滋养层细胞不同，在培养中会分裂。2. 通过测量哇巴因敏感的86Rb+摄取来评估Na(+)-K(+)-ATP酶活性。通过（i）测量[3H]哇巴因结合以及（ii）Northern杂交来测定α-1和β1亚基mRNA的表达，从而确定Na(+)-K(+)-ATP酶的表达。3. 在细胞滋养层细胞分化过程中，Na(+)-K(+)-ATP酶的活性或表达均无显著差异。然而，与培养18小时的细胞滋养层细胞相比，培养66小时的细胞滋养层细胞中α1和β1亚基mRNA的表达显著降低。4. JAr细胞中的Na(+)-K(+)-ATP酶活性和[3H]哇巴因结合均显著高于任何一组细胞滋养层细胞，尽管α1和β1亚基mRNA的表达与培养18小时的细胞滋养层细胞相同。5. 得出的结论是，在细胞滋养层细胞分化过程中，N(+)-K(+)-ATP酶活性和蛋白质表达没有变化，但在转录或转录后水平存在表达变化。