Effects of endogenous nitric oxide induced by 5-fluorouracil and L-Arg on liver carcinoma in nude mice.
Effects of endogenous nitric oxide induced by 5-fluorouracil and L-Arg on liver carcinoma in nude mice.
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DOI：
10.3748/wjg.v13.i46.6249
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发表时间：
2007-12
影响因子：
4.3
通讯作者：
Xiao-Yan Yin;Jun-Mei Jiang;Ji-yong Liu;Ju-ren Zhu
中科院分区：
文献类型：
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作者：
Xiao-Yan Yin;Jun-Mei Jiang;Ji-yong Liu;Ju-ren Zhu
目的研究5-氟尿嘧啶(5-Fu)和L精氨酸(L-Arg)诱导的内源性一氧化氮(NO)对裸鼠人肝癌模型的影响。方法用BEL-7402细胞和生理盐水(NS)、5-FU及5-FU+L-精氨酸建立裸鼠人肝癌模型。测量肿瘤大小。显微镜下观察坏死灶的程度和范围。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法检测肿瘤细胞的凋亡。免疫组织化学方法检测iNOS、P16、Bax的表达。采用化学比色法测定其活性，硝酸还原酶法测定肿瘤组织中一氧化氮(NO)的含量。采用BI2000病理图像分析仪对免疫组织化学结果进行分析。结果5-FU联合L-精氨酸对肿瘤生长有明显抑制作用。生理盐水组、5-FU组和5-FU+L-精氨酸组肿瘤体积变化分别为257.978±59.0 mm、172.232±66.0 mm和91.523±26.7 mm(P<；0.05 5-FU vs 5-FU+L精氨酸组；P<；0.05 NS vs 5-FU+L精氨酸组；P<；0.05，NS vs 5-FU+L精氨酸组)。注射药物后，坏死范围和细胞凋亡指数明显增加。5-FU+L-精氨酸组肿瘤坏死范围最大(c2=15.963，P<0.05)。细胞凋亡指数：NS为17.4%+/-6.19%，5-FU为31.3%+/-12.3%，5-FU+L-Arg为46%+/-15.24%(P<；0.05，5-FU vs 5-FU+L-Arg；P<；0.05，NS vs 5-FU+L-Arg；P<；肿瘤组织中诱导型一氧化氮合酶表达增强，活性增强。NO浓度也明显升高。诱导型一氧化氮合酶光密度、诱导型一氧化氮合酶活性和一氧化氮浓度的F分别为31.693、21.949和33.909。NO浓度与P16、Bax表达呈正相关。相关系数分别为0.764和0.554。结论5-FU联合L-精氨酸对裸鼠移植瘤有抑制作用。其作用机制可能与诱导iNOS合成，提高iNOS活性有关。增加NO的产生，增强细胞凋亡相关基因和抑癌基因的表达。
AIM To study the effects of endogeous nitric oxide induced by 5-fluorouracil (5-FU) and L-arginine (L-Arg) on the human liver carcinoma model in nude mice. METHODS The human liver carcinoma model in nude mice was established with BEL-7402 cells and normal saline (NS), 5-FU and 5-FU + L-Arg injected intraperitoneally. The tumor size was measured. The necrotic degree and range were observed under microscope. The apoptosis of cancer cell was detected by turmina deoxynucleotidyl transferanse mediated dUTP nick end labeling (TUNEL) method. Immunohistochemical method was performed to determine the expression of iNOS, P16, BAX. The chemical colorimetry was used to test the activity and nitrate reductase method was adopted to test the concentration of nitric oxide (NO) in the tumor tissue. The BI2000 pathological image analyzer was used to analyze the result of immunohistochemistry. RESULTS 5-FU combined with L-Arg could inhibit the tumor growth apparently. In NS, 5-FU and 5-FU+L-Arg groups, the changes of tumor volumes were 257.978 +/- 59.0, 172.232 +/- 66.0 and 91.523 +/- 26.7 mm(3), respectively (P < 0.05 5-FU vs 5-FU + L-Arg group; P < 0.05 NS vs 5-FU + L-Arg group; P < 0.05, NS vs 5-FU group). The necrotic range and apoptosis index were significantly increased after the drug injection. The necrotic range was biggest in 5-FU + L-Arg group (c2 = 15.963, P < 0.05). The apoptosis indexes were as follows: NS, 17.4% +/- 6.19%; 5-FU, 31.3% +/- 12.3%; and 5-FU + L-Arg, 46% +/- 15.24% (P < 0.05, 5-FU vs 5-FU + L-Arg; P < 0.05, NS vs 5-FU + L-Arg; P < 0.05, NS vs 5-FU). The expression and activity of iNOS were increased in the tumor tissue. The concentration of NO was also increased. F of optical density of iNOS, iNOS activity and NO concentration are 31.693, 21.949, and 33.909, respectively, P < 0.05. The concentration of NO was related to the expression of P16 and BAX. The correlation coefficient was 0.764 and 0.554. CONCLUSION 5-FU combined with L-Arg can inhibit the growth of tumor in nude mice. The effect may be related to inducing the synthesis and increasing the activity of iNOS. The production of NO is increased, and it can enhance the expression of apoptosis-related gene and antioncogene.