关于冻化疗对G422胶质瘤细胞凋亡及调控机制的影响

　　【摘要】 目的:观察冷冻联合5-Fu对G422胶质瘤化疗敏感性的影响，并探讨其内在的分子机制。方法:建立G422胶质瘤动物模型，随机分组，分别施以化疗、冷冻、冻化疗;TUNEL法用于观察各组在不同时间点细胞凋亡的变化;免疫组化方法检测冷冻后p27kip1蛋白的表达。结果:冷冻与化疗均能引起肿瘤细胞凋亡，冻化疗的联合应用使凋亡率明显增加。冷冻中心区少有p27kip1的阳性表达，冻后周边区表达大为增加。冻后凋亡细胞与p27kip1的表达呈显著正相关(P<0.01) 。结论:冷冻能够提高G422胶质瘤对5-Fu的化疗敏感性，其发生机制可能与冻后p27kip1诱发肿瘤细胞凋亡有关;冻化疗的联合应用较单纯冷冻或化疗治疗恶性胶质瘤效果更为明显。

　　【关键词】 诱发肿瘤细胞 胶质瘤 分子机制 TUNEL染色

　　The effect of cryotherapy on appoptosis and its mechanism in G422 glioma model QIN Jun,WANG Hui，TU Han-Jun,et al.(Department of Neurosurgery,Taihe Hospital,Yunyang Medical College,Shiyan,Hubei 442000)

　　Abstract:Objective To observe the chemosensitivity of glioma treated by cryotherapy to 5-Fu and to explore the potential mechanism. Methods Experimental mice models with G422 glioma was established.Chemotherapy,cryotherapy and cryochemotherapy were performed respectively;TUNEL was used to detect glioma cell apoptosis of each group at different time points;The expression of the p27kip1 gene of the glioma treated by cryotherapy was analyzed by immunohistochemical staining.Results Cryotherapy and 5-Fu could induce apoptosis of G422 glioma,while the combination of cryotherapy and 5-Fu had a remarkably higher ratio of apoptosis.No expression of p27kip1gene was observed in the center of frozen area,the p27kip1 protein increased obviously in the periphery of the frozen area.The expression of p27kip1 was positively correlated with that of the numbers of apoptosis in glioma treated with cryotherapy(P<0.01).Conclusions Cryotherapy could enhance the chemosensitivity of G422 glioma to 5-Fu presumably through modulating the p27kip1 expression pattern;cryochemotherapy showed more potent effect on glioma apoptosis than that of cryotherapy or chemotherapy alone.

　　Key words：Cryochemotherapy;Glioma;Chemosensitivity;p27kip1

　　胶质瘤发病率高，术后易复发，手术切除结合放、化疗效果仍不满意，冻化疗的联合应用作为一种肿瘤治疗模式虽已有报道[1]，但其深入研究的报道较少。本研究利用G422胶质瘤载体模型初步探讨冷冻对其化疗敏感性的影响及其内在的分子机制。

　　1材料与方法

　　1.1瘤株的来源及动物模

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