煤焦沥青对中国仓鼠肺细胞周期和凋亡的影响  

Effect of coal tar pitch on cell cycle and apoptosis of Chinese hamster lung cell

作　　者：马振云[1] 朱纪法[2] 罗宝刚[3] 张明五[4] 

机构地区：[1]河南省三门峡市疾病预防控制中心,472000 [2]郑州大学第一附属医院 [3]河南科技大学第二附属医院 [4]浙江省疾病预防控制中心

出　　处：《职业与健康》2015年第2期184-186,189,共4页Occupation and Health

摘　　要：目的探讨煤焦沥青对中国仓鼠肺(CHL)细胞c-jun、fos和表皮生长因子受体(EGFR)m RNA表达和细胞周期和凋亡的影响。方法常规培养CHL细胞,常规消化细胞后,将细胞接种于6孔板中,待其融合度达到40%左右时,加入含有煤焦沥青的培养液,调整浓度至0、20、40 mg/L,分别于作用24、72 h后提取细胞总RNA,采用半定量RT-PCR技术检测c-jun、fos和EGFR m RNA的表达水平;将等量的细胞接种在50 ml的培养瓶中,待细胞融合度达到40%左右时,加入含煤焦沥青的培养液,调整浓度为0、20、40 mg/L,每组设置6个重复样品,分别加入谷胱甘肽(GSH)和N-乙酰半胱氨酸(NAC),调整浓度至0、2.5、10 mmol/L,作用120 h后收集细胞,采用流式细胞术检测细胞凋亡率和S期细胞的构成比。结果在20 mg/L煤焦沥青作用24 h后,CHL细胞的fos、c-jun和EGFR m RNA的表达水平均高于0 mg/L剂量组,差异均有统计学意义(P<0.05);作用72 h后c-jun和EGFR的表达水平显著高于0 mg/L剂量组(P<0.05)。在40 mg/L剂量组作用24和72 h后,fos、c-jun和EGFR m RNA的表达水平均高于0 mg/L剂量组,差异均有统计学意义(P<0.05)。煤焦沥青作用后,细胞凋亡率降低,在GSH作用下,凋亡率有所增加,NAC作用下的凋亡率增加的幅度更加明显,但以上差异均无统计学意义(P>0.05);20 mg/L的煤焦沥青作用120 h后,S期的细胞构成显著增加(P<0.05);GSH和NAC可以显著抑制S期的细胞构成改变,差异有统计学意义(P<0.05)。结论煤焦沥青可以诱导fos、c-jun和EGFR m RNA的表达水平和S细胞周期的构成增加、抑制细胞凋亡率,而GSH和NAC可以抑制这种诱导效应。[Objective]To explore the effect of coal tar pitch（CTP）on expression of c-jun,fos and epidermal growth factor receptor（EGFR）m RNA,as well as apoptosis and cell cycle of Chinese hamster lung（CHL）cell.[Methods]CHL cells were cultured in the6-well cell culture plate,added culture medium containing CTP of 0,20 and 40 mg/L while the cells fusion reached 40%,and then total RNA was extracted after culturing for 24 hours and 72 hours respectively. The expression levels of c-jun,fos and EGFR m RNA were detected with semi-quantitative RT-PCR. CHL cells of same amounts were cultured in 50 ml culture bottle, and added culture medium containing CTP of 0,20 and 40 mg/L while the cells fusion reached 40%. There were 6 duplicate samples in each group,which were respectively treated with glutathione（GSH）and n-acetylcysteine（NAC）of 0,2.5 and 10 mmol/L for 120 hours, and then the apoptosis rate and constituent ratio of cells in S phase were detected by flow cytometry. [ Results]With stimulation of 20mg/L CTP for 24 hours,the expression levels of c-jun,fos and EGFR m RNA of CHL cells were higher than those in 0 mg/L CTP group, and the differences were statistically significant（P〈 0.05）. After stimulation for 72 hours,the expression levels of c-jun and EGFR m RNA in 20mg/L CTP groups were significantly higher than those in 0 mg/L CTP group（P0.05）. With stimulation of 40mg/L CTP for 24 hours and 72 hours,the expression levels of c-jun,fos and EGFR m RNA were higher than those in 0 mg/L CTP group,and the differences were statistically significant（P0.05）. Apoptosis rate reduced after treating with CTP, while it increased under treatment of GSH,and the amplification of apoptosis rate was more obvious by treating with NAC,but all differences were not statistically significant（P〉0.05）. With stimulation of 20mg/L CTP for 120 hours, constituent ratio of cells in S phase increased significantly（P0.05）,while the change was inhibited by both of GSH and NAC,and the differences were statistically signif

关 键 词：煤焦沥青 中国仓鼠肺细胞 fos、c-jun EGFR 细胞周期 细胞凋亡 

分 类 号：R114[医药卫生—卫生毒理学]