雷帕霉素对体外培养的鼻息肉细胞增殖的影响  

Effect of Rapamycin on the Proliferation of Nasal Polyp Cells in vitro

作　　者：徐婷婷[1,2] 白伟良 谭海燕[1] 王枭维[1] XU Tingting;BAI Weiliang;TAN Haiyan;WANG Xiaowei(Department of Otolaryngology,Shengjing Hospital,China Medical University,Shenyang 110004,China;Department of Otolaryngology,Tieling Central Hospital,Tieling 112000,China)

机构地区：[1]中国医科大学附属盛京医院耳鼻咽喉科,沈阳110004 [2]铁岭市中心医院耳鼻咽喉科,辽宁铁岭112000

出　　处：《中国医科大学学报》2019年第11期978-982,共5页Journal of China Medical University

基　　金：辽宁省科学事业公益研究基金(2013001010);沈阳市科技计划(F11-264-1-07,F13-220-9-21)

摘　　要：目的研究哺乳动物雷帕霉素靶蛋白(mTOR)抑制剂(雷帕霉素)对鼻息肉细胞增殖的影响,分析m TOR信号通路在鼻息肉的发生发展中的作用。方法取鼻息肉患者术中摘除组织进行体外原代细胞培养,流式细胞术鉴定细胞,待细胞生长至80%~90%融合时,加入不同浓度的雷帕霉素(0、3、10、100、300、1 000、3 000、10 000、30 000 ng/mL)培养48 h,用MTT法测鼻息肉细胞增殖情况,运用改良的窛式法计算半抑制浓度(IC50)。结果流式细胞术检测结果显示,对照组阳性细胞率为(4.8±0.9)%,实验组阳性细胞率为(60.9±4.7%),2组差异存在统计学意义(t=26.214,P <0.001);0、3、10、100、300、1 000、3 000、10 000、30 000 ng/mL雷帕霉素组的细胞抑制率分别为0%、(11.6±0.4)%、(21.78±1.2)%、(31.8±4.8)%、(42.8±5.4)%、(82.9±6.5)%、(91.2±6.8)%、(96.2±6.4)%、(98.2±7.1)%。与0 ng/mL雷帕霉素组比较,100 ng/mL和300 ng/mL雷帕霉素组抑制率明显增加(P <0.05);1 000 ng/mL及以上浓度组抑制率均在80%左右,对细胞抑制率增大更明显(均P <0.05),IC50为495.800 ng/mL。结论雷帕霉素可抑制鼻息肉细胞增殖,mTOR信号通路在鼻息肉的发生发展中可能起一定的作用。Objective To study the effect of rapamycin,an inhibitor of mammalian target of rapamycin(mTOR),on the proliferation of nasal polyp cells and to analyze the role of the mTOR signaling pathway in the occurrence and development of nasal polyps. Methods The tissues of patients with nasal polyps were excised for primary cell culture in vitro;these cells were subsequently identified by flow cytometry. When the cells reached 80%-90% confluence,various concentrations of rapamycin(0,3,10,100,300,1 000,3 000,10 000,and 30 000 ng/mL) were added to the culture for 48 h. The proliferation of nasal polyp cells was measured using the MTT assay,and the half maximal inhibitory concentration(IC50) was calculated by the modified Crew method. Results Flow cytometry showed that the positive cell rates in the control group and experimental group were 4.8%±0.9%,and 60.9%±4.7%,respectively;there was a statistically significant difference between these two groups(t = 26.214,P < 0.001). The cell inhibition rates of the 0,3,10,100,300,1 000,3 000,10 000,and 30 000 ng/mL rapamycin groups were 0%,11.60%±0.40%,21.78%±1.20%,31.80%±4.80%,42.80%±5.40%,82.90%±6.50%,91.20%±6.80%,96.20±6.40%,and 98.20±7.10%,respectively. Compared with 0 ng/mL,the inhibition rates of the 100 ng/mL and 300 ng/mL rapamycin groups were significantly increased(P < 0.05). The inhibition rate was about 80% at concentrations above 1 000 ng/mL,and the inhibition rate increased more dramatically with an increased cell concentration(P < 0.05);IC50 was 495.800 ng/mL. Conclusion Rapamycin can inhibit the proliferation of nasal polyp cells,and the mTOR signaling pathway may play a role in the occurrence and development of nasal polyps.

关 键 词：鼻息肉细胞 哺乳动物雷帕霉素靶蛋白抑制剂 雷帕霉素 增殖 

分 类 号：R765[医药卫生—耳鼻咽喉科]