温经膏穴位贴敷诱导肿瘤细胞凋亡及对p53基因的影响

目的 :探讨温经膏穴位贴敷在肺癌治疗中的作用机理。方法 :温经膏穴位贴敷于荷瘤小鼠相当于人体的大椎和肺俞穴 ,观察温经膏对细胞形态、原位末端标记 (TUNEL)及 p53基因的影响。结果 :温经膏贴敷组细胞凋亡指数与CTX组比较无显著性差异 (P >0 .0 5) ,与荷瘤组比较有显著性差异 (P <0 .0 5) ;温经膏贴敷组和CTX组均无p53基因阳性表达及突变 ,贴敷对照组和荷瘤组有p53基因阳性表达及突变。结论 :温经膏能诱导肿瘤细胞凋亡 ,可能有恢复抑癌基因 p53功能的作用

Influence of Acupoint Application of "Wenjing" Plaster on Apoptosis and p53 Gene Expression

Objective: To study mechanisms of acupoint application of "Wenjing" Plaster (温经膏,"Wenjinggao")in treatment of lung cancer. Methods: 40 tumor-bearing mice were randomly and evenly divided into "Wenjinggao" composed of Chansu (蟾酥,Venenum Bufonis), Caowu (草乌,Radix Aconiti Kusnezoffii), Baijiezi (白芥子,Semen Sinapis Albae), Bingpian (冰片,Borneolum), etc group, common plaster control group, cytoxan (CTX) group and cancer model group. Common plaster (composed of vehicle) and "Wenjinggao" were respectively applied to "Dazhui" (GV 14) and "Feishu" (BL 13) for 2 hours every day, beginning from the second day on after inoculation of tumor that was performed by subcutaneous injection (right subaxillary) of cancer cell suspension (S180, 0.2 mL/mouse) to establish cancer-bearing model. In the mice of CTX group, CTX (0.4 mg/case) was given (i.p.) to the mice from the 3rd day on after the inoculation. Ten days later, the mice were killed for taking out the tumor tissues which were sectioned to be observed under light microscope and transmission electron microscope. Apoptosis was assayed by using TdT-mediated dUTP notch end labeling (TUNEL) technique and p53 gene expression was detected with polymerase chain reaction restriction fragment length polymorphismin method. Results: In "Wenjinggao" group, electron microscopic results showed a loose arrangement of the cells, sparse cytoplasm, severe margination of the nuclear chromatin with some chromatinorrhexis (apoptosis), while in model group, no apparent apoptosis phenomenon (dying cells and died corpuscles) mentioned above was found. TUNEL results displayed that the apoptosis indexes of "Wenjinggao" group, CTX group and model group were 3.06%± 1.31%, 3.22%±1.78% and 0.22%±0.09% respectively, no significant difference was found between the former two groups (P>0.05), while there was a significant difference between "Wenjinggao" group and model group in the apoptosis indexes(P<0.05). In "Wenjinggao" group and CTX group, no positive p53 expression strip (117 bp) and mutation (the 248th and 249th codons) were found, while in control group and model group, these positive strip and mutation existed clearly. Conclusion: "Wenjinggao" can evoke apoptosis and renew functions of p53 (cancer-inhibitory gene).