CAJ | 학술논문

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。
목적관찰체외배양적종류세포대거서세포염증인자--종류배사인자-α（TNF-α）적유도작용；초보탐색최알삼서（CAI）대종류유도적거서세포중 TNF-α석방적영향。방법이용 transwell 장치，건립 Lewis 폐암세포（LLC）여 RAW264.7거서세포공배양체계，채용형광정량 PCR방법화 ELISA 방법분별분석 RAW264.7중 TNF-α적표체화석방；용 LLC 조건배양기（LCM）유도 RAW264.7，동시급여 CAI 처리，채용 CCK-8방법분석 LCM 화 CAI대 RAW264.7활력적영향，채용 ELISA 방법분석 CAI 대유도후적 RAW264.7중 TNF-α석방적영향。 　　결과여 LLC 공배양24 h 가현저증가 RAW264.7중TNF-α상대표체량[단독배양 vs 공배양，（1.00±0.12）vs （2.23±0.17），P <0.01]화석방[단독배양 vs 공배양，（65.21±12.76）vs （143.92±19.22）pg/ml，P<0.05]；LCM 화 CAI 재1 h 화4 h 대 RAW264.7활력몰유영향，CAI 가이현저억제 LCM 대 RAW264.7중 TNF-α석방적유도（4 h，P<0.01）。 　　결론 LLC 종류미배경가이유도 RAW264.7중 TNF-α적표체증가；CAI 대저충유도적억제사기재종류배경중표현출일정적항염작용，가능시기항종류작용적궤제지일。
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.