氢氧化钠抑制原位移植性肝癌生长机制研究  

The mechanism of sodium hydroxide-inhibited growth of hepatocellular carcinoma in orthotopic intrahepatic model

作　　者：李勇[1] 郑筠 党亚正[1] 陆婉玲[1] 刘军[1] 齐涛[1] 

机构地区：[1]解放军第323医院肿瘤中心,陕西西安710054 [2]乌鲁木齐市中医医院,新疆乌鲁木齐830000

出　　处：《西部医学》2015年第7期978-980,985,共4页Medical Journal of West China

基　　金：陕西省自然科学基金(2010JM4053;2014JM4121)

摘　　要：目的探讨氢氧化钠溶液瘤内注射对原位肝癌的生长抑制作用,研究抵抗肝癌生长的临床机制。方法获得稳定表达红色荧光蛋白DsRed的人肝癌细胞系SMMC-7721,进行不同浓度氢氧化钠溶液刺激,并使用原位末端标记法(TdT-mediated dUTP nick end labeling,TUNEL)检测细胞凋亡。建立裸鼠原位肝癌模型,于第7天进行不同浓度氢氧化钠溶液原位瘤内注射,利用小动物活体荧光成像技术观察肿瘤生长情况。利用免疫组织化学法检测残存肿瘤组织中VEGF表达,并使用TUNEL法检测瘤体内细胞凋亡。结果氢氧化钠溶液刺激肝癌细胞后,显著增加细胞凋亡比例。成功建立裸鼠原位肝癌模型并进行氢氧化钠溶液注射。在荧光成像系统下,激发光波长570nm,发射波长610nm,于细胞接种后第21天观测到发红色荧光的肿瘤。取肝脏进行原位瘤体积测定,发现与生理盐水瘤内注射相比,氢氧化钠显著抑制了原位瘤生长并引起瘤组织坏死。氢氧化钠抑制了瘤体中VEGF的表达,并增加了TUNEL阳性细胞(即凋亡细胞)比例。结论氢氧化钠瘤内注射有效抑制裸鼠原位肝癌生长,这可能是通过抗血管生成作用和诱导凋亡作用实现的。Objective To investigate the effects of sodium hydroxide on hepatocellular carcinoma xenograft in an orthotopic intrahepatic model.Methods SMMC-7721 cells were transfected with a vector to establish a DsRed stably expressed cell line,named as 7721-DsRed.Cells were incubated with different doses of sodium hydroxide or normal saline followed by detection of apoptosis by TdT-mediated dUTP nick end labeling（TUNEL）.Nude mice bearing 7721-DsRed tumors in livers were injected with different doses of sodium hydroxide or normal saline at day 7post injection.In vivo DsRed signal was monitored using a Kodak Image Station 4000 MM PRO Digital Imaging System.The expression of VEGF and apoptosis in the periphery of the target were detected by immunohistochemistry and TUNEL assay respectively.Results Stimulation of sodium hydroxide significantly increased the percentage of apoptotic HCC cells.The orthotopic intrahepatic model of hepatocellular carcinoma was successfully established.DsRed signal of in situ tumors was detected at the indicated time points（day 21）（excitation and emission wavelengths were 570 and 610nm respectively）.Compared to control group,sodium hydroxide induced necrosis,significantly inhibited the size of in situ tumors and increased the weight of mice.Moreover,the expression of VEGF was suppressed and the percentage of apoptotic cells in tumors was increased.Conclusion By intratumoral injection with sodum hydroxide in the situ HCC tumors of orthotopic intrahepatic model,the growth of tumors was effectively suppressed.The underlying mechanism might be the inhibition of angiogenesis and the induction of apoptosis.

关 键 词：肝细胞肝癌 氢氧化钠 VEGF TUNEL 

分 类 号：R735.7[医药卫生—肿瘤]