Zhang L, Ruan X, Gu M, Mueck A O
Department of Gynecological Endocrinology, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing, China.
Department of Women's Health, University Women's Hospital and Research Center for Women's Health, University of Tuebingen, Tuebingen, Germany.
Climacteric. 2022 Oct;25(5):467-475. doi: 10.1080/13697137.2022.2029837. Epub 2022 Feb 9.
This study aimed to find evidence that progesterone receptor membrane component 1 (PGRMC1) promotes estradiol (E2) + norethisterone (NET)-induced breast cancer proliferation through activation of the phosphatidylinositol-3-kinase (PI3K)-AKT pathway.
PGRMC1-mediated breast cancer cellular proliferation and phosphorylation of PGRMC1 were studied using wild-type (hemagglutinin [HA]-tagged) MCF-7 cells, which were stably transfected with expression vector containing HA (MCF-7-HA cells), PGRMC1 (MCF-7-PGRMC1 cells) and Ser181 point mutated PGRMC1 (MCF-7-PGRMC1-S181A cells). Bioinformatics, cell proliferation, western blot, isobaric tags for relative and absolute quantitation (iTRAQ)-based RNA sequencing, real-time quantitative polymerase chain reaction (RT-qPCR) and cell cycle assays were performed to indicate the function of PGRMC1 and its possible mechanisms in breast cancer.
NET + E2 elicited a significant proliferation in MCF-7-Vec at 10 M and 10 M, respectively. MCF-7-PGRMC1 did increase the phosphorylation of AKT or ERK, which can be blocked by treatment with casein kinase 2 (CK2) inhibitor quinalizarin or in MCF-7-PGRMC1-S181A cells. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that the PI3K-AKT pathway is upregulated in MCF-7-PGRMC1 cells. Importantly, upregulation of the PI3K-AKT pathway mainly through promotion of cell cycle regulation strongly promoted cell proliferation in MCF-7-PGRMC1 cells.
CK2 is involved in phosphorylation of PGRMC1 at S181. The mechanism for the action of PGRMC1 for mediating proliferative progestogen effects obviously starts with promotion cell cycle regulation, and then activation of the PI3K-AKT pathway.
本研究旨在寻找证据，证明孕激素受体膜成分1（PGRMC1）通过激活磷脂酰肌醇-3-激酶（PI3K）-AKT途径促进雌二醇（E2）+炔诺酮（NET）诱导的乳腺癌增殖。
使用野生型（血凝素[HA]标记）MCF-7细胞研究PGRMC1介导的乳腺癌细胞增殖和PGRMC1的磷酸化，这些细胞用含有HA的表达载体（MCF-7-HA细胞）、PGRMC1（MCF-7-PGRMC1细胞）和Ser181点突变的PGRMC1（MCF-7-PGRMC1-S181A细胞）进行稳定转染。进行生物信息学、细胞增殖、蛋白质印迹、基于相对和绝对定量的等压标签（iTRAQ）的RNA测序、实时定量聚合酶链反应（RT-qPCR）和细胞周期分析，以表明PGRMC1在乳腺癌中的功能及其可能机制。
NET + E2分别在10 μM和10 μM时引起MCF-7-Vec细胞显著增殖。MCF-7-PGRMC1确实增加了AKT或ERK的磷酸化，这可以通过用酪蛋白激酶2（CK2）抑制剂喹哪嗪处理或在MCF-7-PGRMC1-S181A细胞中被阻断。京都基因与基因组百科全书（KEGG）分析显示，PI3K-AKT途径在MCF-7-PGRMC1细胞中上调。重要的是，PI3K-AKT途径的上调主要通过促进细胞周期调控强烈促进MCF-7-PGRMC1细胞的增殖。
CK2参与PGRMC1在S181位点的磷酸化。PGRMC1介导增殖性孕激素作用的机制显然始于促进细胞周期调控，然后激活PI3K-AKT途径。
