摘要
目的研究尼古丁在口腔鳞状细胞癌细胞增殖和诱导凋亡中的作用，初步探讨尼古丁对口腔鳞状细胞癌发生的作用机制。方法采用甲基噻唑基四唑法、膜联蛋白V-异硫氰酸荧光素及碘化丙啶双染法和2’，7’一二氯荧光黄双乙酸盐荧光探针法检测不同浓度（0．1、1、10μmol／L）尼古丁作用相同时间（48h）和相同浓度（1μmol／L）尼古丁作用不同时间（24、48、72h），对口腔鳞状细胞癌SCCl5细胞活力、细胞凋亡和细胞内活性氧含量的影响，以1μmol／L尼古丁为对照组；应用酶联免疫吸附测定、免疫荧光方法检测lμmol／L尼古丁作用不同时间后口腔鳞状细胞癌SCCl5细胞内核因子KBDNA结合活性及核因子KB表达变化。结果不同浓度尼古丁处理SCCl5细胞48h，流式细胞术检测各组细胞内滑性氧水平分别为（98．24±O．04）％、（98．50±O．06）％及（98．61±0．07）％，均较对照组[（96．01±0．58）％]显著增加（P=0．000）；同时各浓度尼古丁处理组细胞生长被促进，且呈浓度依赖性，0．1、1、10μmol／L尼古丁组细胞4值分别为2．19±0．08、2．20±0．11及2．38±0．08，均显著高于对照组（1．93±0．13）（P〈0．05）。1ixmol／L尼古丁处理组的SCCl5细胞凋亡率显著高于对照组及0．1、10μmol／L处理组（P=0．000）。1Iμmol／L尼古丁作用于SCCl5细胞72h细胞生长被显著抑制，与对照组相比差异有统计学意义（P=0．022）；1μmol／L尼古丁作用于SCCl5细胞24h后诱导SCCl5细胞凋亡最显著且显著高于尼古丁作用48h和72h组（P：0．000），各组细胞核内代表DNA结合活性的4值分别为1．509、1．093、0．746，与对照组（4值为0．544）相比核因子KBDNA结合活性均有不同程度增高，提示核因子kB从胞质向胞核中转移，其中尼古丁作用24h活性最高。结论尼古丁能促进口腔鳞状细胞癌SCCl5细胞增殖并诱导少量细胞凋�
Objective To investigate the role of nicotine on the proliferation and cell apoptosis in SCC15 oral squamous cell carcinoma cells. Methods The growth, apoptosis, reactive oxygen species （ROS） level and nuclear factor kappalight-chain-enhancer of activated B cell（NF-KB） DNA binding activity were detected in SCC15 oral cancer cell using methly thiazolyl tetrazolium assay, flow cytometry, enzyme linked immunosorbent assay. Results In SCC15 cells treated with nicotine for 48 h at different concentrations （0. l, 1, 10 μmol/L） ROS level was （98.24 ± 0. 04）%, （98.50 ± 0. 06）% , （98.61 ± O. 07 ） % , respectively, which were significantly higher than in control groups [ （ 96. 01 ± 0. 58 ） % , P = 0. 000] and the A value for cell growth was 2. 19 ±0. 08,2. 20 ±0. 11 and 2. 38 ±0. 08, respectively, which were significantly higher than in control groups（ I. 93 ＋0. 13） （P 〈0. 05）. Only 1 p.mol/L nicotine induced significantly higher cell apoptosis than in other groups （P = 0. 000）. Cell growth was inhibited in SCC15 cells treated with 1 p.mol/L nicotine for 72 h, which had statistically significant difference compared with control （ P = 0. 022）. Cell apoptosis rate in 1 μmol/L nicotine treated groups for 24 h was significantly higher than 48 h and 72 h（P = 0. 000）. NF-KB expression in the nucleus were increased in SCC15 cells treated with 1 p±mol/L nicotine for 24, 48 and 72 h and the A value for NF-kB DNA binding activity was 1. 509,1. 093 and 0. 746, respectively,which were higher than in control group（0. 544）. Conclusions Nicotine induced SCC15 cell growth and apoptosis, which maybe by NF-KB signal pathway activated in oral cancer cells.
出处
《中华口腔医学杂志》
CAS
CSCD
北大核心
2012年第4期233-237,共5页
Chinese Journal of Stomatology
基金
北京市自然科学基金（7102065）
