A Novel Role for Progesterone and Progesterone Receptor Membrane Component 1 in Regulating Spindle Microtubule Stability During Rat and Human Ovarian Cell Mitosis
A Novel Role for Progesterone and Progesterone Receptor Membrane Component 1 in Regulating Spindle Microtubule Stability During Rat and Human Ovarian Cell Mitosis
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DOI：
10.1095/biolreprod.110.088385
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发表时间：
2011-04-01
影响因子：
3.6
通讯作者：
Peluso, John J.
中科院分区：
文献类型：
--
作者：
Lodde, Valentina;Peluso, John J.
本研究旨在评估黄体酮（P4）和黄体酮受体膜组分1 （PGRMC1）在调节自发永生化颗粒细胞（SIGCs）和卵巢癌细胞SKOV-3细胞有丝分裂中的作用。由于PGRMC1已经在人类有丝分裂纺锤体的蛋白质中被检测到，我们推测P4和PGRMC1可能通过微管依赖的过程影响有丝分裂。目前的研究证实，无论是P4治疗还是PGRMC1抗体转染，SIGC的生长都会减慢。在这两种情况下，细胞增殖减慢都伴随着有丝分裂细胞百分比的增加，这与p4诱导的细胞周期M期延长一致。此外，P4增加了纺锤体微管的稳定性，这是通过对冷却反应的β -微管蛋白分解率来评估的。P4还增加了SKOV-3细胞的纺锤体微管稳定性。这种效应被这些细胞中PGRMC1的耗尽所模仿。重要的是，与pgrmc1缺失的SKOV-3细胞相比，P4并没有增加微管的稳定性。免疫荧光分析显示PGRMC1分布于纺锤体和中期中心体。进一步原位接近连接实验显示PGRMC1与β -微管蛋白相互作用。综上所述，这些结果表明P4通过增加有丝分裂纺锤体的稳定性来抑制卵巢细胞的有丝分裂。此外，P4的作用似乎依赖于PGRMC1在有丝分裂纺锤体中的功能。
The present studies were designed to assess the roles of progesterone (P4) and Progesterone Receptor Membrane Component 1 (PGRMC1) in regulating mitosis of spontaneously immortalized granulosa cells (SIGCs) and ovarian cancer cells, SKOV-3 cells. Because PGRMC1 has been detected among the proteins of the human mitotic spindle, we theorized that P4 and PGRMC1 could affect mitosis through a microtubule-dependent process. The present study confirms that SIGC growth is slowed by either P4 treatment or transfection of a PGRMC1 antibody. In both cases, slower cell proliferation was accompanied by an increased percentage of mitotic cells, which is consistent with a P4-induced prolongation of the M phase of the cell cycle. In addition, P4 increased the stability of the spindle microtubules, as assessed by the rate of beta-tubulin disassembly in response to cooling. Also, P4 increased spindle microtubule stability of SKOV-3 cells. This effect was mimicked by the depletion of PGRMC1 in these cells. Importantly, P4 did not increase the stability of the microtubules over that observed in PGRMC1-depleted SKOV-3 cells. Immunofluorescent analysis revealed that PGRMC1 is distributed to the spindle apparatus as well as to the centrosomes at metaphase. Further in situ proximity ligation assay revealed that PGRMC1 interacted with beta-tubulin. Taken together, these results suggest that P4 inhibits mitosis of ovarian cells by increasing the stability of the mitotic spindle. Moreover, P4' s actions appear to be dependent on PGRMC1' s function within the mitotic spindle.