Expression of the p53 and Wilms' tumor suppressor genes in the rat ovary: gonadotropin repression in vivo and immunohistochemical localization of nuclear p53 protein to apoptotic granulosa cells of atretic follicles.
Expression of the p53 and Wilms' tumor suppressor genes in the rat ovary: gonadotropin repression in vivo and immunohistochemical localization of nuclear p53 protein to apoptotic granulosa cells of atretic follicles.
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DOI：
10.1210/endo.136.4.7895650
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发表时间：
1995-04
期刊：
影响因子：
4.8
通讯作者：
K. I. Tilly;Subhadra Banerjee;Partha P. Banerjee;Jonathan L. Tilly
中科院分区：
文献类型：
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作者：
K. I. Tilly;Subhadra Banerjee;Partha P. Banerjee;Jonathan L. Tilly
我们先前已经证明促性腺激素介导的卵巢颗粒细胞凋亡抑制与几个细胞死亡相关基因表达的变化有关，包括bcl-2基因家族成员（bcl-2，bax和bcl-x）。最近，p53肿瘤抑制基因的产物，一种被报道在调节细胞增殖和死亡中起关键作用的蛋白质，已被证明直接调节bcl-2和bax基因的转录活性。此外，p53的作用可以通过与另一种肿瘤抑制蛋白（Wilms肿瘤抑制基因（WT-1）的产物）的协同相互作用来放大。基于我们对bcl-2相关因子在调节颗粒细胞凋亡中的潜在作用的鉴定以及报道的p53作为性腺外细胞中bcl-2和bax基因转录的调节剂的功能，目前的研究是为了确定p53和WT-1基因的表达和促性腺激素的调节，并研究颗粒细胞凋亡是否与肿瘤水平升高有关。抑制基因表达。从未成熟（27日龄）大鼠卵巢制备的总RNA的北方印迹分析显示存在单个p53信使RNA（mRNA）转录物（2.0个酶）和多个WT-1信息（1.8、3.5和7.5个酶）。皮下注射10 IU马CG（eCG）的未成年大鼠2天后，p53和WT-1 mRNA水平分别降至生理盐水处理对照水平的71 +/- 9%（P < 0.05）和46 +/- 3%（P < 0.05）。eCG治疗对肿瘤抑制基因表达的抑制与凋亡颗粒细胞和闭锁卵泡数量的显着减少有关。此外，免疫组化分析显示，p53蛋白只定位于细胞核的凋亡颗粒细胞的闭锁卵泡，并在体内治疗后，与eCG p53免疫染色降低到无法检测的水平。为了进一步评估颗粒细胞凋亡是否与肿瘤抑制基因表达增加有关，我们分析了无血清培养诱导的有腔卵泡闭锁中p53和WT-1 mRNA的水平。（摘要截短至400字）
We have previously demonstrated that the gonadotropin-mediated inhibition of apoptosis in ovarian granulosa cells is linked to changes in the expression of several cell death-related genes, including members of the bcl-2 gene family (bcl-2, bax, and bcl-x). Recently, the product of the p53 tumor suppressor gene, a protein reported to play a critical role in regulating cell proliferation and death, has been shown to directly modulate the transcriptional activity of the bcl-2 and bax genes. In addition, the actions of p53 may be amplified through a cooperative interaction with another tumor suppressor protein, the product of the Wilms' tumor suppressor gene (WT-1). Based on our identification of a potential role for bcl-2-related factors in regulating granulosa cell apoptosis and the reported function of p53 as a regulator of bcl-2 and bax gene transcription in extragonadal cells, the present studies were conducted to determine whether the p53 and WT-1 genes are expressed and gonadotropin regulated in the rat ovary and to investigate whether granulosa cell apoptosis is linked to elevated levels of tumor suppressor gene expression. Northern blot analysis of total RNA prepared from immature (27-day-old) rat ovaries revealed the presence of a single p53 messenger RNA (mRNA) transcript (2.0 kilobases) and multiple WT-1 messages (1.8, 3.5, and 7.5 kilobases). Subcutaneous injection of immature rats with 10 IU equine CG (eCG) reduced the levels of p53 and WT-1 mRNA to 71 +/- 9% (P < 0.05) and 46 +/- 3% (P < 0.05), respectively, of saline-treated control levels after 2 days. The inhibition of tumor suppressor gene expression by eCG treatment was associated with a marked reduction in the number of apoptotic granulosa cells and atretic follicles. Furthermore, immunohistochemical analysis revealed that p53 protein was localized exclusively to nuclei of apoptotic granulosa cells of atretic follicles, and that p53 immunostaining was reduced to undetectable levels after in vivo treatment with eCG. To further evaluate whether granulosa cell apoptosis is linked to increased expression of tumor suppressor genes, we analyzed levels of p53 and WT-1 mRNA in antral follicles induced to undergo atresia in vitro by serum-free culture.(ABSTRACT TRUNCATED AT 400 WORDS)