氩氦刀对T739小鼠LA795皮下移植瘤坏死及细胞凋亡的影响  

Effect of Argon-Helium cryoablation on necrosis and cell apoptosis of subcu-taneously transplanted tumor of LA795 in T739 mice

作　　者：温居一[1] 段蕴铀[1] 邹曰坤[1] 冯华松[1] 聂舟山[1] 

机构地区：[1]海军总医院呼吸内科,北京100037

出　　处：《第四军医大学学报》2007年第19期1742-1745,共4页Journal of the Fourth Military Medical University

基　　金：全军"十一五"医药卫生科研项目(06MB021)

摘　　要：目的：研究氩氦刀冷冻消融对T739小鼠LA795肺腺癌皮下移植瘤坏死和细胞凋亡的影响.评价氩氦刀冷冻治疗非小细胞肺癌的可行性及疗效.方法：建立T739小鼠LA795肺腺癌皮下移植瘤动物模型.氩氦刀冷冻后,光学显微镜观察冷冻坏死消融规律;末端标记法（TUNEL）原位检测周边冷冻损伤区带细胞凋亡,免疫组织化学法检查Bc1-2和Bax表达;WesternBlot检测冰球周边冷冻损伤区Caspase-3表达及聚腺苷二磷酸核苷聚合酶（PARP）剪切活性.结果：在冷冻中心区,细胞死亡以坏死为主,坏死呈现＂双峰＂效应：氩氦刀术后3h出现第1次坏死高峰,坏死区域面积/肿瘤面积达（47±5.51）%;术后4d出现第2次坏死峰,坏死区域面积/肿瘤面积达（68±4.69）%.在周边冷冻损伤区,光镜形态学检查及TUNEL染色可观察到明确的细胞凋亡,术后8~16h凋亡达高峰,凋亡率为（71±6.41）%.免疫组化显示,Bcl-2蛋白表达在冷冻前后无明显变化,bax蛋白在冷冻后其表达上调.Western Blot结果显示：冷冻后冰球周边损伤区带Caspase-3和PARP剪切、激活.结论：氩氦刀通过坏死和凋亡两种途径达到有效的消融.冷冻消融可能通过Caspase途径诱导周边损伤区带细胞凋亡.其凋亡机制有Bax蛋白参与,而与Bcl-2无关.AIM： To study the effect of Argon-Helium cryoablation on necrosis and apoptosis of subcutaneously transplanted tumor of the lung adenocarcinoma LA795 in T739 mice in vivo, thus to evaluate the feasibihy and therapeutic efficacy of argonhelium cryoablation in treatment of Non-small cell lung cancer （NSCLC）. METHODS： Animal model of subcutaneously transplanted tumor of the lung adenocarcinoma LA795 in T739 mice was establishd. After argon-helium cryoablation, necrosis and ablation laws were studied under light microscope. Cell apoptosis in peripheral cryoablation zone was detected by in-situ end labeling （TUNEL）. Bcl-2 and Bax expression were detected by immunohistochemical SABC procedures. The cleavage and activation of Caspase 3 and PARP in peripheral cryoablation zone was detected by Western Blot. RESULTS ： In central area of cryoablation, necrosis dominated in cell death, showing double peak efficacy. The first necrosis peak occurred at 3 h after argon-helium cryoablation （ area of necrotic area/area of tumor =47% ） ; the 2nd necrosis peak occurred at 4 days after cryoablation （ area of necrotic area/area of tumor = 68% ）. In peripheral cryoablation zone, definite cell apoptosis was observed by morphological examination under light microscope and TUNEL staining, and it reached the peak at 8 - 16 h after cryoablation. Immunohistochemical results showed no obvious change in Bcl-2 protein expression before and after cryoablation. Bax protein expression was up-regulated after crvoablation. Western Blot results showed cleavage and activation of Caspase-3 and PARP in peripheral cryoablation zone around ice ball after the treatment. CONCLUSION： Argon-Helium-based cryoablation is an efficient technique to induce cell death either by necrosis or by apoptosis. Cryoablation might induce cell apoptosis in peripheral cryoablation zone through Caspase pathway. Bax protein may be involved in apoptosis mechanism, but it is irrelevant to Bcl-2.

关 键 词：Endocare氩氦刀 冷冻消融 腺癌 凋亡 

分 类 号：R73-36[医药卫生—肿瘤]