氧化砷诱导食管癌细胞凋亡中一氧化氮的变化  

Alterations of nitric oxide in apoptosis of esophageal carcinoma cells induced by arsenite

作　　者：沈忠英[1] 沈文英[2] 陈铭华 洪超群 沈健 

机构地区：[1]汕头大学医学院肿瘤病理研究室 [2]汕头大学医学院化学教研室 [3]肿瘤医院中心实验室,广东省汕头市515031

出　　处：《世界华人消化杂志》2000年第10期1101-1104,共4页World Chinese Journal of Digestology

摘　　要：目的一氧化氮 NO 已被认为是生理和病理状态下一种重要生物分子.以氧化砷(As_2O_3)诱导食管癌细胞(SHEEC1)凋亡过程中检测细胞 NO,NO 合成酶(NOS)和一氧化氮合成酶mRNA(NOSmRNA),探索 NO 和细胞凋亡的关系.方法用 As_2O_3(5 μmol·L^(-1)和10μmol·L^(-1))诱导 SHEEC1凋亡,在加药后2,4,8,16,24h 取样检查,用分光光密度法检测细胞培养液的 NO 含量;用免疫组化检测诱导型 NO 合成酶(NOSⅡ);分子原位杂交(ISH)检测 NOSmRNA;实验终点细胞作透射电镜(TEM)检查细胞凋亡.结果在 As_2O_3诱导下,SHEEC1从 NO 基础量(0.68×10^(-2)μmol·L^(-1))逐渐增加,至加药后16h 达最高(2.38×10^(-2)μmol·L^(-1));NOSⅡ在加药后4h～16h 呈阳性反应;NOSmRNA 杂交信号位于胞质,4h 杂交点小而少,以后增加明显.24h,TEM可见 SHEEC1出现细胞凋亡.结论 As_2O_3 可诱导 SHEEC1释放 NO 和细胞凋亡,实验开始至16h NO 量不断增加.NOSⅡ表达和 NOSmRNA 转录上调.提出 NO 可能是 As_2O_3诱导细胞凋亡的介导和作用因子.Nitric oxide (NO) has been known as an important molecule in critical physiological and pathological conditions.To determine NO,NO synthase (NOS) and NOS mRNA of the esophageal carcinoma cells (SHEEC1) in apoptotic process induced by As_2O_3 and to explore the relationship between NO and apoptosis. METHODS The apoptosis of the cell line (SHEEC1) was induced by arsenite (As_2O_3,5 end 10μmol·L^(-1)).In the process,at 2,4,8,16 and 24 h after administration of As_2O_3,NO production in culture medium was detected quantitatively by spectrophotometry;NOSⅡ was detected by immunohistochemistry and NOS mRNA was detected by insitu hybridization (ISH).The cells at endpoint of the experiment were examined under transmitted electron microscope (TEM) for apoptosis. RESULTS The amount of NO released from SHEFC1 were increased from the basal condition (0.68×10^(-2)μmol·L^(-1) upto the high level (2.38×10^(-2)μmol· L^(-1)) at the 16th hour.The increment of NOS Ⅱ was found after administration of As_2O_3;The intracytoplesmic ISH signals of NOSmRNA in small size was found firstly at 4 h,and they became highly predominant after ward.Apoptotic changes of SHEEC1 occured at 24 h under TEM. CONCLUSION After administration of As_2O_3,NO released from cultured SHEEC1 cells was detected with increasing amount until 16h.The expression of NOS Ⅱ and transcription of NOSmRNA are upregulated.The present findings suggest that NO may be a mediator and effect factor of apoptosis induced by As_2O_3.

关 键 词：食管肿瘤 细胞凋亡 免疫组织化学 

分 类 号：R735.1[医药卫生—肿瘤]