xCT knockdown in human breast cancer cells delays onset of cancer-induced bone pain
xCT knockdown in human breast cancer cells delays onset of cancer-induced bone pain
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DOI：
10.1177/1744806918822185
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发表时间：
2019-01-08
期刊：
影响因子：
3.3
通讯作者：
Singh, Gurmit
中科院分区：
文献类型：
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作者：
Ungard, Robert G.;Linher-Melville, Katja;Singh, Gurmit
骨中的癌症产生许多严重的症状，包括损害患者功能状态、生活质量和生存的疼痛。这种疼痛的来源是多方面的，包括肿瘤细胞分泌的因子。恶性细胞通过氧化应激相关的胱氨酸/谷氨酸反向转运体系统x（C）（-）释放神经递质和细胞信号分子谷氨酸，该系统可转运胱氨酸用于谷胱甘肽的合成和胱氨酸/半胱氨酸氧化还原循环。系统x（C）（-）的药理学抑制已在小鼠模型中成功减少和延迟癌症疼痛相关行为的发作。本研究描述了在人乳腺癌细胞系MDA-MB-231中稳定的siRNA诱导的功能性跨膜系统x（C）（-）亚基xCT（SLC 7A 11）敲低的发展。在转录物、蛋白质和功能水平上验证克隆的xCT敲低。对代表性克隆进行RNAseq以全面检查响应于xCT敲低的转录细胞特征，鉴定与癌症疼痛相关的多种差异调节因子，包括神经生长因子、白细胞介素-1和集落刺激因子-1。小鼠股内接种，并记录疼痛相关的行为，包括负重，机械撤回，和肢体使用。相对于对照细胞，植入xCT敲低癌细胞的动物显示出直到伤害性行为发作的延迟。这些结果增加了大量证据，表明通过抑制系统x（C）（-）转运蛋白减少骨骼癌症的谷氨酸释放可能会减少与骨转移相关的严重和顽固性疼痛。
Cancers in the bone produce a number of severe symptoms including pain that compromises patient functional status, quality of life, and survival. The source of this pain is multifaceted and includes factors secreted from tumor cells. Malignant cells release the neurotransmitter and cell-signaling molecule glutamate via the oxidative stress-related cystine/glutamate antiporter, system x(C)(-), which reciprocally imports cystine for synthesis of glutathione and the cystine/cysteine redox cycle. Pharmacological inhibition of system x(C)(-) has shown success in reducing and delaying the onset of cancer pain-related behavior in mouse models. This investigation describes the development of a stable siRNA-induced knockdown of the functional trans-membrane system x(C)(-) subunit xCT (SLC7A11) in the human breast cancer cell line MDA-MB-231. Clones were verified for xCT knockdown at the transcript, protein, and functional levels. RNAseq was performed on a representative clone to comprehensively examine the transcriptional cellular signature in response to xCT knockdown, identifying multiple differentially regulated factors relevant to cancer pain including nerve growth factor, interleukin-1, and colony-stimulating factor-1. Mice were inoculated intrafemorally and recordings of pain-related behaviors including weight bearing, mechanical withdrawal, and limb use were performed. Animals implanted with xCT knockdown cancer cells displayed a delay until the onset of nociceptive behaviors relative to control cells. These results add to the body of evidence suggesting that a reduction in glutamate release from cancers in bone by inhibition of the system x(C)(-) transporter may decrease the severe and intractable pain associated with bone metastases.