雌激素调控奶牛乳腺上皮细胞凋亡及生长周期作用机制的研究

doi:10.16431/j.cnki.1671-7236.2019.09.010

摘要/Abstract

摘要： 试验旨在研究雌激素对奶牛乳腺上皮细胞（BMECs）凋亡及生长周期的影响。通过添加MAPK/ERK信号通路阻断剂探索雌激素调控BMECs凋亡及生长周期具体的作用机制，采用流式细胞仪检测细胞凋亡及周期的变化情况，实时荧光定量PCR检测Bcl-2、Caspase3及CyclinD1基因mRNA的表达丰度。结果显示，对照组BMECs凋亡率极显著低于BMECs+PD98059、BMECs+E₂+PD98059组（P<0.01），Bcl-2 mRNA表达丰度极显著高于BMECs+PD98059组（P<0.01），Caspase3 mRNA表达丰度显著低于BMECs+PD98059组（P<0.05）；对照组细胞比例在G1期显著高于BMECs+E₂组（P<0.05），极显著低于BMECs+E₂+PD98059组（P<0.01），S期细胞比例极显著高于BMECs+PD98059、BMECs+E₂+PD98059组（P<0.01），G2期细胞比例极显著低于BMECs+PD98059、BMECs+E₂+PD98059组（P<0.01）；对照组CyclinD1 mRNA的表达丰度极显著高于BMECs+PD98059组（P<0.01）；BMECs+E₂+PD98059组的Bcl-2 mRNA的表达量极显著高于BMECs+PD98059组（P<0.01），Caspase3 mRNA的表达量显著低于BMECs+PD98059组（P<0.05）。结果表明，MAPK/ERK信号通路参与BMECs的增殖及细胞生长周期调节的过程，且雌激素可通过MAPK/ERK信号通路抑制BMECs的凋亡，MAPK/ERK信号通路可能参与由雌激素调控的细胞生长周期的进程。

关键词: 雌激素; 乳腺上皮细胞; MAPK/ERK; 凋亡; 周期; 机制

Abstract: The purpose of this experiment was to study the effect of estrogen on apoptosis and growth cycle of bovine mammary epithelial cells (BMECs).The specific mechanism of estrogen regulating apoptosis and growth cycle of BMECs was explored by adding MAPK/ERK signaling pathway blockers.Flow cytometry was used to detect apoptosis and cycle changes.Real-time PCR was used to detect the expression abundance of Bcl-2,Caspase3 and CyclinD1 genes.The results showed that the apoptotic rate of BMECs in control group was extremely significantly lower than that in BMECs+PD98059 and BMECs+E₂+PD98059 groups (P<0.01),the expression abundance of Bcl-2 was extremely significantly higher than that in BMECs+PD98059 group (P<0.01),and the expression abundance of Caspase3 was significantly lower than that in BMECs+PD98059 group (P<0.01);In control group,the proportion of G1 phase cells was significantly higher than that in BMECs+E₂ group (P<0.05),and was extremely significantly lower than that in BMECs+E₂+PD98059 group (P<0.01),the proportion of S phase cells was significantly higher than that in BMECs+PD98059 and BMECs+E₂+PD98059 groups (P<0.01),the proportion of G2 phase cells was extremely significantly lower than that of BMECs+PD98059 and BMECs+E₂+PD98059 groups (P<0.01);The expression abundance of CyclinD1 gene was extremely significantly higher than that in BMECs+PD98059 group (P<0.01);The expression of Bcl-2 gene in BMECs+E₂+PD98059 group were extremely significantly higher than that in BMECs+PD98059 group (P<0.01),and the expression of Caspase3 was extremely significantly lower in BMECs+PD98059 group than that in BMECs+PD98059 group (P<0.01).The results showed that the MAPK/ERK signaling pathway was involved in the regulation of proliferation and cell growth cycle of mammary epithelial cells,and estrogen could inhibit the apoptosis of mammary epithelial cells through the MAPK/ERK signaling pathway.The MAPK/ERK signaling pathway might be involved in the process of cell growth cycle regulated by estrogen.

Key words: estrogen; mammary epithelial cells; MAPK/ERK; apoptosis; cycle; mechanism

中图分类号:

S823

武开乐, 库西塔别克·买买提依不拉音, 马静, 肖凡, 余雄, 邵伟. 雌激素调控奶牛乳腺上皮细胞凋亡及生长周期作用机制的研究[J]. 中国畜牧兽医, 2019, 46(9): 2575-2582.

WU Kaile, KUXITABIEKE·Maimaitiyibulayin, MA Jing, XIAO Fan, YU Xiong, SHAO Wei. Study on the Mechanism of Estrogen Regulating Apoptosis and Growth Cycle of Bovine Mammary Epithelial Cells[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(9): 2575-2582.

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