GPR30/HOTAIR通过雌激素非基因组效应调节乳腺癌细胞增殖及毛蕊异黄酮的干预作用

Estrogen plays a key role in the development of cancer. Previous studies have demonstrated that ER positive endothelail cells mediates angiogenesis through both classic genomic and rapid non-genomic mechanisms. Rapid non-genomic mechanisms is becoming a focus. Some study revealed that GPR30 and HOTAIR were overexpressed specifically in ER-positive cells. However, the cross talk of GPR30 and HOTAIR in.cells was unclear. Our previous studies suggested that calycosin of phytoestrogen had effects on the proliferation and apoptosis of the breast cancer cells in vitro and in vivo, finding that the effects might be related to the level of HOTAIR and the activation of PI3K/Akt. Furthermore, our previous studies confirmed that GPR30 downregulated the level of HOTAIR. Based on the above information, we hypothesize that the cross talk between GPR30 and HOTAIR may also be involved in the antiproliferative effect induced by calycosin in breast cancer cells. By using cell counting, MTT assay, flow cytometry, luciferase activity assay, Western blot, qPCR, Xenograft tumor growth and immunohistochemistry, we will identify upregulated or downregulated HOTAIR and GPR30 in breast cancer cells are able to interfere with PI3K/Akt signaling pathways in vitro and in vivo. Accordingly, further research should be carried out to elucidate the relationship among calycosin of phytoestrogen, HOTAIR and GPR30, which may provide experimental foundation for phytoestrogen's future clinical use for breast cancer.

雌激素在肿瘤的发生发展中占有重要的地位，而雌激素非基因组效应日益受到重视。有研究提出GPR30与HORAIR可通过非基因组效应干预PI3K/Akt通路，影响乳腺癌细胞增殖凋亡，但它们之间相互作用的研究未见报道。课题组前期研究表明异黄酮类植物雌激素—毛蕊异黄酮下调多种乳腺癌细胞HOTAIR水平，同时通过HOTAIR-PI3K/Akt通路影响乳腺癌细胞增殖。我们的预实验结果同时显示干预GPR30可影响乳腺癌细胞的HOTAIR水平。据此，我们推测毛蕊异黄酮通过干预HOTAIR与GPR30交互式对话，影响下游信号通路来发挥作用。本项目拟通过细胞生物学、分子生物学、荷瘤裸鼠等实验，探讨雌激素非基因组效应关键靶点GPR30/HOTAIR在体内外的信号级联，重点论证毛蕊异黄酮对HOTAIR、GPR30交互式对话及下游PI3K/Akt通路蛋白的影响，为异黄酮类植物雌激素预防和治疗乳腺癌寻找新的靶点。

雌激素在肿瘤的发生发展中占有重要的地位，而雌激素非基因组效应日益受到重视。有研究提出GPR30与HORAIR可通过非基因组效应干预PI3K/Akt通路，影响乳腺癌细胞增殖凋亡，但它们之间相互作用的研究未见报道。课题组前期研究表明异黄酮类植物雌激素—毛蕊异黄酮下调多种乳腺癌细胞HOTAIR水平，同时通过HOTAIR-PI3K/Akt通路影响乳腺癌细胞增殖。我们的预实验结果同时显示干预GPR30可影响乳腺癌细胞的HOTAIR水平。据此，我们推测毛蕊异黄酮通过干预HOTAIR与GPR30交互式对话，影响下游信号通路来发挥作用。本项目拟通过细胞生物学、分子生物学、荷瘤裸鼠等实验，探讨雌激素非基因组效应关键靶点GPR30/HOTAIR在体内外的信号级联，重点论证毛蕊异黄酮对HOTAIR、GPR30交互式对话及下游PI3K/Akt通路蛋白的影响，为异黄酮类植物雌激素预防和治疗乳腺癌寻找新的靶点。