摘要
目的观察体外培养的肿瘤细胞对巨噬细胞炎症因子——肿瘤坏死因子-α(TNF-α)的诱导作用;初步探索羧胺三唑(CAI)对肿瘤诱导的巨噬细胞中TNF-α释放的影响。方法利用transwell装置,建立Lewis肺癌细胞(LLC)与RAW264.7巨噬细胞共培养体系,采用荧光定量PCR方法和ELISA方法分别分析RAW264.7中TNF-α的表达和释放;用LLC条件培养基(LCM)诱导RAW264.7,同时给予CAI处理,采用CCK-8方法分析LCM和CAI对RAW264.7活力的影响,采用ELISA方法分析CAI对诱导后的RAW264.7中TNF-α释放的影响。结果与LLC共培养24 h可显著增加RAW264.7中TNF-α相对表达量[单独培养vs共培养,(1.00±0.12)vs(2.23±0.17),P<0.01]和释放[单独培养vs共培养,(65.21±12.76)vs(143.92±19.22)pg/ml,P<0.05];LCM和CAI在1 h和4 h对RAW264.7活力没有影响,CAI可以显著抑制LCM对RAW264.7中TNF-α释放的诱导(4 h,P<0.01)。结论 LLC肿瘤微环境可以诱导RAW264.7中TNF-α的表达增加;CAI对这种诱导的抑制使其在肿瘤环境中表现出一定的抗炎作用,可能是其抗肿瘤作用的机制之一。
Objective To investigate the tumor necrosis factor-α（TNF-α） expression and secretion enhancement in macrophages induced by tumor cells in vitro, as well as the inhibitory effect of carboxyamidotriazole （CAI） on TNF-α induction in tumor-educated macrophages. Methods The Lewis lung carcinoma （LLC） cells and RAW264.7 macrophages were co-cultured with the transwell inserts, LLC conditioned medium （LCM） was also used to induce RAW264.7 with or without CAI treatment. Real time RT-PCR and ELISA methods were applied to analyze the TNF-αexpression and secretion in RAW264.7, respectively. CCK-8 was applied to determine the effect of LLC （LCM） and CAI on the viability of RAW264.7. Results TNF-αexpression [cultured alone vs co-cultured, （1.00 ± 0.12） vs （2.23 ± 0.17）, P〈0.01] and secretion [cultured alone vs co-cultured, （65.21 ± 12.76） vs （143.92 ± 19.22） pg/ml, P〈0.05] were greatly increased in RAW264.7 after being co-cultured with LLC for 24 hours. LCM and CAI did not influence the viability of RAW264.7 at 1 hour and 4 hour, while CAI inhibited TNF-αsecretion in RAW264.7 induced by LCM （4 h, P〈0.01）. Conclusion LLC tumor environment can greatly enhance the TNF-α expression in RAW264.7. CAI inhibits this induction significantly and shows anti-inflammation activity in tumor environment, which may contribute to its anti-cancer effects.
出处
《中国医药生物技术》
2014年第2期124-128,共5页
Chinese Medicinal Biotechnology
基金
"十二五"国家科技重大专项(2014ZX09507003-003)
国家自然科学基金(81201728)
高等学校博士学科点专项科研基金新教师类(20121106120019)
关键词
肿瘤坏死因子
α
巨噬细胞
共培养
羧胺三唑
Tumor necrosis factor-alpha
Macrophages
Co-culture
Carboxyamidotriazole
