力尔凡对膀胱癌抑制作用的试验研究

Experimental study on the inhibitory effect of Lifein on growth of bladder tumor

目的研究力尔凡对膀胱癌的抑制作用,探讨其可能的作用机制。方法将不同浓度的力尔凡作用于膀胱癌EJ细胞,使用MTT法检测细胞增殖情况,利用流式细胞仪(FCM)检测细胞周期体内试验用16只4～6周龄的Balb/c裸鼠种植膀胱癌EJ细胞构建荷瘤模型,随机分为力尔凡组和生理盐水组,使用酶联免疫分析法(ELISA)检测瘤体组织冲洗液中IL-2和TNF-α的水平,对瘤体组织进行HE染色以观察肿瘤组织坏死情况,并采用免疫组化法检测IL-12和p53的表达水平。结果力尔凡对EJ细胞具有抑制增殖的作用,并在一定范围呈出时间和浓度依赖性;力尔凡(100μg/ml)作用24小时、48小时后,G_2/M期细胞增多,S期细胞减少体内试验瘤体组织上清液用酶联免疫分析法检测,力尔凡组IL-2和TNF-α水平明显增高;HE染色可见力尔凡组肿瘤组织出现坏死,免疫组织化学法示力尔凡组p53表达明显降低,IL-12表达明显增高。结论力尔凡可以改变膀胱癌EJ细胞周期分布,抑制细胞增殖,并能增加瘤体组织IL-2、TNF-α、IL-12等细胞因子含量,使瘤体p53表达减少,从而抑制甚至杀伤膀胱癌细胞。

Objective To study the inhibitory effects of Lifein on growth of bladder tumor and the underlying mechanisms. Method EJ cells were treated with Lifein at different concentrations. MTT cell proliferation assay was used to observe the inhibitory effect. The cell cycle and cell apoptosis were detected by FCM. In the in vivo experiment, 16 Balb/ c nude mice of 4 -6 weeks-old were used to build bladder cancer model, which were randomized into Lifein group （6 mg/ kg） and control group （saline）. Cytokines （IL-2, TNF-α） were detected by ELISA. Immunocytochemical staining was used to analyze the expression of p53, and IL-12 protein. Result Lifein inhibited the proliferation of EJ cells by dosedependent and time-dependent manner. EJ cells were treated with 100 μg/ml Lifein for 24 and 48 hours, cells in GJM phase were increased in cell cycle, but S-phase cells decreased. In the in vivo experiment, the level of IL-2 and TNF-α increased in Lifein group. Results of immunocytochemical staining showed that Lifein down-regulated the expression of p53 protein, while up-regulated the IL-12 expression. Conclusion Lifein can change the distribution of cell cycle, inhibit the proliferation of bladder cancer EJ cell, increase the level of IL-2, TNF-α and IL-12, decrease the expression of p53 protein. Thus, it can inhibit and even kill bladder cancer ceils.