αO-Conotoxin GeXIVA Inhibits the Growth of Breast Cancer Cells via Interaction with α9 Nicotine Acetylcholine Receptors
αO-Conotoxin GeXIVA Inhibits the Growth of Breast Cancer Cells via Interaction with α9 Nicotine Acetylcholine Receptors
复制标题
α O-芋螺毒素 GeXIVA 通过与 α 9 尼古丁乙酰胆碱受体相互作用抑制乳腺癌细胞的生长
DOI：
10.3390/md18040195
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发表时间：
2020-04-01
期刊：
影响因子：
5.4
通讯作者：
Luo, Sulan
中科院分区：
文献类型：
--
作者：
Sun, Zhihua;Bao, Jiaolin;Luo, Sulan
含有α 9的烟碱乙酰胆碱受体（nAChR）由于其在乳腺癌中的高表达特异性而日益成为新的肿瘤靶点。α O-芋螺毒素GeXIVA是α 9 α 10 nAChR的有效拮抗剂。然而，GeXIVA对乳腺癌细胞的抗肿瘤作用仍不清楚。采用Cell Counting Kit-8法研究不同剂量GeXIVA作用于乳腺癌MDA-MD-157细胞和人正常乳腺上皮细胞后的细胞活力。采用流式细胞仪检测GeXIVA对乳腺癌细胞周期阻滞和凋亡的影响。通过伤口愈合试验分析迁移能力。Western blot（WB）、实时荧光定量PCR（QRT-PCR）和流式细胞术检测α 9-nAChR的表达。使用CRISPR/Cas9技术建立了α 9-nAChR亚基敲除（KO）的稳定MDA-MB-157乳腺癌细胞系。GeXIVA能够显著抑制乳腺癌MDA-MB-157细胞的增殖并促进其凋亡。此外，乳腺癌MDA-MB-157细胞的增殖被GeXIVA抑制，其通过下调α 9-nAChR引起细胞周期阻滞。GeXIVA还可抑制MDA-MB-157细胞的迁移。这表明GeXIVA诱导α 9-nAChR表达下调，并且由于α 9-nAChR缺失，MDA-MB-157 α 9-nAChR KO细胞系的生长也受到抑制。GeXIVA在体外抑制乳腺癌细胞MDA-MB-157细胞的生长，并且可能在消除α 9-nAChR的机制中发生。
The alpha 9-containing nicotinic acetylcholine receptor (nAChR) is increasingly emerging as a new tumor target owing to its high expression specificity in breast cancer. alpha O-Conotoxin GeXIVA is a potent antagonist of alpha 9 alpha 10 nAChR. Nevertheless, the anti-tumor effect of GeXIVA on breast cancer cells remains unclear. Cell Counting Kit-8 assay was used to study the cell viability of breast cancer MDA-MD-157 cells and human normal breast epithelial cells, which were exposed to different doses of GeXIVA. Flow cytometry was adopted to detect the cell cycle arrest and apoptosis of GeXIVA in breast cancer cells. Migration ability was analyzed by wound healing assay. Western blot (WB), quantitative real-time PCR (QRT-PCR) and flow cytometry were used to determine expression of alpha 9-nAChR. Stable MDA-MB-157 breast cancer cell line, with the alpha 9-nAChR subunit knocked out (KO), was established using the CRISPR/Cas9 technique. GeXIVA was able to significantly inhibit the proliferation and promote apoptosis of breast cancer MDA-MB-157 cells. Furthermore, the proliferation of breast cancer MDA-MB-157 cells was inhibited by GeXIVA, which caused cell cycle arrest through downregulating alpha 9-nAChR. GeXIVA could suppress MDA-MB-157 cell migration as well. This demonstrates that GeXIVA induced a downregulation of alpha 9-nAChR expression, and the growth of MDA-MB-157 alpha 9-nAChR KO cell line was inhibited as well, due to alpha 9-nAChR deletion. GeXIVA inhibits the growth of breast cancer cell MDA-MB-157 cells in vitro and may occur in a mechanism abolishing alpha 9-nAChR.