The expression of Annexin A1 and A5 mRNA by gonadotropin-releasing hormone in LβT2 gonadotrope cells
The expression of Annexin A1 and A5 mRNA by gonadotropin-releasing hormone in LβT2 gonadotrope cells
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DOI：
10.1507/endocrj.ej21-0397
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发表时间：
2021-10-12
影响因子：
2
通讯作者：
Kawaminami, Mitsumori
中科院分区：
文献类型：
--
作者：
Murata, Takuya;Chiba, Shuichi;Kawaminami, Mitsumori
在 L beta T2 促性腺激素细胞中分析促性腺激素释放激素 (GnRH) 对膜联蛋白 A1 (ANXA1) 和 A5 (ANXA5) mRNA 表达的刺激。定量聚合酶链式反应结果显示，GnRH 类似物 (GnRHa) 刺激 ANXA1 和 A5 mRNA 的表达，并在孵育 12 小时时达到峰值；然而，ANXA1 mRNA 受到极大刺激（60 倍）。免疫细胞化学分析证实了这些发现。 GnRH 拮抗剂可抑制 GnRHa 的作用。蛋白激酶 C (PKC) 激活剂（12-O-十四烷酰佛波醇-13-乙酸酯；TPA）可显着增加 ANXA1 和 A5 mRNA 水平，但二丁酰 cAMP 则不会显着增加 ANXA1 和 A5 mRNA 水平。 GnRHa 刺激的 ANXA1 和 A5 mRNA 诱导被 PKC (GF109203) 和 MEK 抑制剂 (PD98059) 抑制。 TPA以剂量依赖性方式增加ANXA1和A5 mRNA表达（1 nM至10 mu M），而ANXA1增加的程度要大得多。用 10 nM 或 1 muM TPA 刺激后，ANXA1 和 A5 mRNA 水平在 6 小时时增加。 1 muM TPA 处理中的 ANXA1 mRNA 水平高于 10 nM TPA 处理，而与 10 nM TPA 相比，1 muM TPA 没有显示出对 ANXA5 mRNA 的进一步刺激。这些结果清楚地表明，ANXA1 mRNA表达与ANXA5一样受到GnRH通过PKC的刺激，并且ANXA1的反应远大于ANXA5。这些膜联蛋白与 ANXA1 在 GnRH 促性腺激素作用中的重要作用之间存在密切关系。
Gonadotropin-releasing hormone (GnRH) stimulation of annexin A1 (ANXA1) and A5 (ANXA5) mRNA expression was analyzed in L beta T2 gonadotrope cells. Quantitative polymerase chain reaction results showed that a GnRH analog (GnRHa) stimulated the expression of both ANXA1 and A5 mRNA with a peak at 12 h of incubation; however, ANXA1 mRNA was extremely stimulated (60 folds). Immunocytochemical analysis confirmed these findings. A GnRH antagonist inhibited the effect of GnRHa. ANXA1 and A5 mRNA levels were significantly increased by protein kinase C (PKC) activator (12-O-Tetradecanoylphorbol-13-acetate; TPA), but not by dibutyryl cAMP. GnRHa-stimulated induction of ANXA1 and A5 mRNA was inhibited by PKC (GF109203) and MEK inhibitors (PD98059). TPA increased ANXA1 and A5 mRNA expression in a dose-dependent manner (1 nM to 10 mu M), while the extent of the increase was much greater in ANXA1. After stimulation with 10 nM or 1 mu M TPA, ANXA1 and A5 mRNA levels were increased at 6 h. ANXA1 mRNA levels were higher in the 1 mu M TPA than in the 10 nM TPA treatment, whereas 1 mu M TPA did not show further stimulation of ANXA5 mRNA compared to 10 nM TPA. These results clearly show that ANXA1 mRNA expression is stimulated by GnRH through PKC like ANXA5, and the response of ANXA1 is much larger than that of ANXA5. A close relationship between these annexins and a significant role for ANXA1 in GnRH action at gonadotropes is suggested.