尼古丁对人肺腺癌细胞和人大细胞肺癌细胞增殖和凋亡的影响

Effects of nicotine on the proliferation and apoptosis of NCI-H1975 cells and NCI-H460 cells

目的观察不同浓度尼古丁对人肺腺癌(NCI-H1975)细胞和人大细胞肺癌(NCI-H460)细胞增殖和凋亡的影响。方法分别取处于对数生长期的NCI-H1975细胞和NCI-H460细胞,调整细胞密度为1×104/ml,分别暴露于含终浓度为0(对照)、1、10、100、1 000、10 000 nmol/L尼古丁的低血清培养基培养72 h;或者分别暴露于含终浓度为0(对照)、1μmol/L尼古丁的低血清培养基培养24、48、72 h,采用CCK-8法检测细胞活性。分别取处于对数生长期的NCI-H1975细胞和NCI-H460细胞,调整细胞密度为1×104/ml,分别暴露于含终浓度为40μmol/L顺伯的低血清培养基24 h,诱导细胞发生凋亡;将诱导后的NCI-H1975细胞、NCI-H460细胞分别暴露于含终浓度为0(对照)、1、10、100、1 000、10 000 nmol/L尼古丁的低血清培养基培养72 h,或者分别暴露于含终浓度为0(对照)、1μmol/L尼古丁的低血清培养基培养24、48、72 h。采用Annexin V/PI染色法检测细胞凋亡率。结果与对照组相比,各浓度尼古丁分别染毒NCI-H1975细胞、NCI-H460细胞72 h后的细胞存活率均增加,差异有统计学意义(P<0.05)。且随着尼古丁染毒浓度的升高,NCI-H1975细胞、NCIH460细胞暴露72 h后的细胞存活率呈明显的先上升后下降趋势。与对照组相比,1μmol/L尼古丁作用NCI-H1975细胞、NCI-H460细胞24、48、72 h后的细胞存活率均较高,差异有统计学意义(P<0.05)。且随着染毒时间的延长,1μmol/L尼古丁暴露NCI-H1975细胞、NCI-H460细胞的细胞存活率呈明显的上升趋势。40μmol/L顺铂作用于NCI-H1975细胞、NCI-H460细胞24 h后,其细胞凋亡率显著升高,与空白组相比,差异有统计学意义(P<0.05)。与对照组相比,各浓度尼古丁作用于顺铂诱导后的NCI-H1975细胞、NCI-H460细胞72 h后,其细胞凋亡率均降低,差异有统计学意义(P<0.05);且随着尼古丁染毒浓度的升高,顺铂诱导后的NCI-H1975细胞、NCI-H460细胞暴露72 h后的细胞凋亡率呈明显的下降的趋势。与对照组相比,1μmol/L尼古丁作用于顺铂诱导后的NCI-H1975细胞、NCI-H460细胞24、48、72 h后的细胞凋亡率均降低,差异有统计学意义(P<0.05);且随着染毒时间的延长,1μmol/L尼古丁暴露顺铂诱导后NCI-H1975细胞、NCIH460细胞的凋亡率呈明显的下降趋势。结论尼古丁可促进NCI-H1975细胞和NCI-H460细胞增殖并抑制细胞凋亡。

Objective To understand the effects of nicotine on the proliferation and apoptosis of human lung adenocarcinoma cells（NCI-H1975） and human large cell lung cancer cells（NCI-H460）. Methods NCI-H1975 and NCI-H460 cells in logarithmic phase with the cell density of 1 ×104/ml were used, treated with nicotine at the doses of 0（control）,1,10,100,1 000,10 000 nmol/L low serum culture medium respectively for 72 h,or treated with nicotine at the doses of 0（control）,1 μmol/L of low serum culture medium for 24,48 and 72 h respectively;The proliferation was detected by CCK-8technique. NCI-H1975 and NCI-H460 in logarithmic phase at the density of 1×104/ml,treated with 40 μmol/L cisplatin of low serum culture medium for 24 h,then 0（control）,1,10,100,1 000 and 10 000 nmol/L nicotine of low serum culture medium for72 h,or 0（control）,1 μmol/L nicotine of low serum culture medium for 24,48,72 h,and the apoptosis was detected by Annexin V/PI staining. Results Compared with the control group,NCI-H1975 cells and NCI-H460 cells viabilities increased significantly after 72 h treatment with nicotine（P〈0.05）. With the increased concentrations of nicotine exposed,NCI-H1975 cells and NCI-H460 cells proliferations were decreased after first then increased. Compared with the control group,in 1 μmol/L nicotine group exposed for 24,48 and 72 h,the cells viabilities increased significantly（P〈0.05） with a time-dependent manner.In 40 μmol/L cisplatin group exposed for 24 h,the apoptosis rates increased significantly（P〈0.05）. Compared with the control group, the apoptosis rates of cisplatin-treated NCI-H1975 cells and NCI-H460 cells treated with nicotine for 72 h decreased significantly（P〈0.05）,with a dose-dependent manner. Compared with the control group, the apoptosis rates of cispatin-treated NCI-H1975 cells and NCI-H460 cells treated with 1 μmol/L nicotine for 24,48 and 72 h decreased significantly（P〈0.05）,with a time-dependent manner. Conclusion Nicotine may promote NCI-H1975 cells and NCI-H460 cells proliferation and inhibit cells apoptosis.