谷氨酰胺合成酶GS通过增强抗凋亡能力降低TNBC细胞对紫杉醇的敏感性

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摘要 目的：三阴性乳腺癌（triple-negative breast cancer,TNBC）具有耐药性强的特点,因此,患者会对临床上最常用的化疗药物紫杉醇敏感性降低,从而导致治疗效果不佳。谷氨酰胺合成酶（glutamine synthetase,GS）是合成谷氨酰胺所需的关键代谢酶,其高表达可诱导卵巢癌细胞对化疗药物降低敏感性。但是,GS是否影响TNBC细胞对紫杉醇的敏感性及其可能机制仍不清楚。本研究的目的是探讨GS是否影响TNBC细胞对紫杉醇的敏感性及其机制。方法：使用质粒转染和慢病毒感染分别构建过表达和敲低GS的TNBC细胞系。通过Western blot、细胞活力检测、克隆形成实验和细胞凋亡检测分析GS在TNBC细胞中对紫杉醇敏感性的影响。使用qRT-PCR检测过表达GS的细胞中药物代谢酶、药物转运蛋白、抗凋亡蛋白的表达。结果：GS在乳腺癌细胞中高表达,GS^low（MDA-MB-231）细胞对紫杉醇的敏感性高于GS^high（MDA-MB-468）细胞。过表达GS使TNBC细胞对紫杉醇的敏感性降低、克隆能力增强、抗凋亡能力增加,敲低GS可以增加紫杉醇对TNBC细胞的杀伤效果;过表达GS的TNBC细胞中,抗凋亡蛋白BAG3的表达显著升高。结论：在TNBC细胞中,高表达GS促进细胞增殖,使TNBC细胞凋亡减少,克隆形成能力增强;GS可能是通过上调BAG3的表达增强抗凋亡能力从而降低TNBC细胞对紫杉醇的敏感性。

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	朱建余
	夏弘卓
	郑卓萌
	彭小宁
	邓锡云
	付淑君

关键词 ：三阴性乳腺癌, 谷氨酰胺合成酶, 紫杉醇, 药物敏感性

Abstract：Objective Triple-negative breast cancer (TNBC) is characterized by strong drug resistance. So patients will be less sensitive to paclitaxel, which is the most commonly used chemotherapeutic drug in the clinic, resulting in poor treatment efficacy. Glutamine Synthetase (GS) is a key metabolic enzyme needed to synthesize glutamine, and its high expression can induce ovarian cancer cells to reduce their sensitivity to chemotherapeutic drugs. However, whether GS affects the sensitivity of TNBC cells to paclitaxel and its possible mechanism are still unclear. The purpose of this study was to investigate whether GS affects the sensitivity of TNBC cells to paclitaxel and its mechanism. Methods TNBC cell lines overexpressing and knocking down GS were constructed by plasmid transfection and lentivirus infection, respectively. The effect of GS on paclitaxel sensitivity in TNBC cells was analyzed by Western blot analysis, cell viability assay, colony formation assay and apoptosis detection. qRT-PCR was used to detect the expression of drug metabolic enzymes, drug transporters, and anti-apoptotic proteins in cells overexpressing GS. Results GS was highly expressed in breast cancer cells. The sensitivity of GS^low cells (MDA-MB-231) to paclitaxel was higher than that of GS^high cells (MDA-MB-468). Overexpression of GS decreased the sensitivity of TNBC cells to paclitaxel, enhanced the ability of clone formation and anti-apoptosis, knocking down GS could increase the killing effect of paclitaxel on TNBC cells, and the expression of anti-apoptotic protein BAG3 was significantly increased in TNBC cells overexpressing GS. Conclusions In TNBC cells, the overexpression of GS promotes cell proliferation, reduces apoptosis, and enhances the ability of clone formation in TNBC cells. The anti-apoptotic effect of GS was mediated by up-regulating the expression of BAG3, thus reducing the sensitivity of TNBC cells to paclitaxel.

Key words： triple negative breast cancer glutamine synthetase paclitaxel drug sensitivity

收稿日期: 2023-05-16

中图分类号:

R737.9

基金资助:国家自然科学基金（82103342,82173374）

通讯作者: 付淑君,E-mail：shujunfu2020@hunnu.edu.cn;邓锡云,E-mail：dengxiyunmed@hunnu.edu.cn

引用本文:

朱建余, 夏弘卓, 郑卓萌, 彭小宁, 邓锡云, 付淑君. 谷氨酰胺合成酶GS通过增强抗凋亡能力降低TNBC细胞对紫杉醇的敏感性[J]. 湖南师范大学学报(医学版), 2023, 20(3): 10-15. ZHU Jianyu, XIA Hongzhuo, ZHENG Zhuomeng, PENG Xiaoning, DENG Xiyun, FU Shujun. Glutamine Synthetase reduces the sensitivity of TNBC cells to paclitaxel by enhancing anti-apoptosis ability. HuNan ShiFan DaXue XueBao(YiXueBan), 2023, 20(3): 10-15.

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http://yxb.hunnu.edu.cn/CN/ 或 http://yxb.hunnu.edu.cn/CN/Y2023/V20/I3/10

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