氩氦刀对T739小鼠LA795皮下移植瘤坏死及细胞凋亡的影响_医学论文.docVIP

氩氦刀对T739小鼠LA795皮下移植瘤坏死及细胞凋亡的影响_医学论文 氩氦刀对T739小鼠LA795皮下移植瘤坏死及细胞凋亡的影响_医学论文 作者：温居一，段蕴铀，邹曰坤，冯华松，聂舟山 【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和Bax表达；Western Blot检测冰球周边冷冻损伤区Caspase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3 h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4 d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8～16 h凋亡达高峰，凋亡率为（71±6.41）％. 免疫组化显示，Bcl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无关. 【关键词】 Endocare氩氦刀；冷冻消融；腺癌；凋亡 　　【Abstract】 AIM: To study the effect of Argons of subcutaneously transplanted tumor of the lung adenocarcinoma LA795 in T739 mice in vivo, thus to evaluate the feasibilty and therapeutic efficacy of argonhelium 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). Bc1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