氯化两面针碱对人鼻咽癌细胞生长和转移的抑制作用

Study on inhibition of nitidine chloride on human NPC cell growth and metastases

目的:探讨氯化两面针碱(nitidine chloride,NC)对表皮生长因子受体酪氨酸激酶(EGFR-TPK)的抑制作用,并阐明其对人鼻咽癌CNE-2细胞增殖和侵袭的抑制作用机制。方法:不同浓度氯化两面针碱加入到人鼻咽癌CNE-2细胞中,利用ELISA法研究了氯化两面针碱对EGFR-TPK活性的影响,利用溴化3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑(MTT)法测定氯化两面针碱对CNE-2细胞的抑制作用,Transwell法检测氯化两面针碱对CNE-2细胞侵袭的抑制作用,采用间接荧光标记法测定细胞内活性氧(ROS)水平的变化;酶标仪法检测氯化两面针碱对细胞凋亡蛋白Caspase-3活性的影响,凝胶迁移或电泳迁移率实验测定CNE-2细胞中NF-κB活性。结果 :氯化两面针碱对EGFR-TPK IC50为(32.53±3.43μmol/L)和CNE-2细胞生长IC50为(9.65±1.13)μmol/L都具有良好的抑制能力,氯化两面针碱能够诱导肿瘤细胞侵袭能力降低;在100μmol/L浓度的氯化两面针碱引起的ROS百分率达到了(76.39±5.32)%;与空白组相比,氯化两面针碱能显著提高CNE-2细胞中凋亡蛋白Caspase 3活性;氯化两面针碱能够下调NF-κB的表达。结论:氯化两面针碱通过多靶向降低EGFR-TPK活性、下调NF-κB的表达和激活Caspase 3活性的方式,对人鼻咽癌细胞的侵袭和增殖产生抑制作用。

Objective： To investigate the inhibiting effect of nitidine chloride on EGFR-TPK and the mechanism of nitidine chloride on CNE-2. Methods ： ELISA was used to detect the activity of EGFR-TPK activity. MTT method was used to detect the inhibitive rate of NC on CNE-2 cell. Transwell method was used to detect the inhibitive rate of NC on CNE-2 cell invasion. Fluorescence marker method was used to detect the ROS level in cell. Caspase-3 activity was detected by enzyme standard instrument. NF-κB activity in CNE-2 cell was detected by electrophoretic mobility shift assays. Results： NC had good inhibiting ability on EGFR-TPK（ IC50 （32.53 ± 3.43） μmol/L） and CNE-2 cell growth （IC50 （9.65±1.13）pznol/L）. Compared with conetrol group, cell invasive ability of NC significantly reduced（ P 〈0.05 ） ; When NC was in a dose of 100μmol/L, ROS was （76.39 ± 5.32） %, the difference was statistically significant compared with control group （ P 〈 0.05 ）. NC can obviously increase Caspase 3 activity in CNE-2 cell（ P 〈 0.05 ）. NC can lower NF-κB expression. Conclusion： NC can lower the EGFR-TPK activity, down-regulated NF-κB expression, activate Caspase 3, and inhibit human NPC cell growth and metastases.