双硫仑联合Cu抑制伯基特淋巴瘤Raji细胞移植瘤生长的动物实验

Inhibition effect of disulfiram combined with Cu on the growth of human Burkitt lymphoma cell xenografts in nude mice

目的 探讨双硫仑（DS）联合铜（Cu）对人伯基特淋巴瘤Raji细胞裸鼠移植瘤生长的抑制作用及分子机制.方法 每只裸鼠给予全身2GyX线照射2 min,于背部近腋下处皮下接种对数生长期Raji细胞1×10^7个,建立Raji细胞裸鼠移植瘤模型;18只造模成功的裸鼠按数字表法随机分为三组：对照组、DS组、DS/Cu组;观察不同用药组裸鼠移植瘤生长的情况,计算肿瘤体积,绘制肿瘤生长曲线,比较三组移植瘤体积和质量.免疫印迹法检测不同用药组处理后移植瘤组织中p-JNK、c-jun蛋白表达.结果 DS组、DS/Cu组和对照组裸鼠移植瘤的体积分别为（67.71±2.15） mm3、（33.35±7.74） mm3、（246.40±42.88） mm3,肿瘤质量分别为（43.35±4.22）mg、（18.05±2.88）mg、（91.30±27.27）mg,三组间差异具有统计学意义（F=27.579,P=0.000;F=16.369,P=0.000）.多重比较结果显示,DS组、DS/Cu组裸鼠移植瘤的体积和质量与对照组比较差异均具有统计学意义（均P＜ 0.05）;DS/Cu组移植瘤体积和质量均低于DS组（均P＜ 0.05）.DS组和DS/Cu组抑瘤率分别为63.48％和80.24％.DS和DS/Cu处理后移植瘤细胞出现了凋亡的改变,DS/Cu组更加明显.DS和DS/Cu处理后p-JNK、c-jun蛋白表达均上调,其中DS/Cu组上调更加明显.结论 DS/Cu可抑制伯基特淋巴瘤Raji细胞裸鼠移植瘤生长,其机制可能与活化JNK通路有关.

Objective To investigate the effects of disulfiram （DS） combined with Cu on the human Burkitt lymphoma cell xenografts in nude mice.Methods Burkitt lymphoma xenograft was established by subcutaneous injection of Raji cell into nude mice after 2 Gy whole body X-irradiation （1×10^7 Raji cells were resuspended in 200 μl saline）.18 bearing tumor mice were randomly divided into control group,DS group and DS/Cu group.During the experiment,the effects of DS/Cu on the nude mice with tumors were examined,including the tumor volumes,weights and the growth curves of xenograft tumor.Histopathological examination of tumor tissue was observed with optical microscape.The protein expression levels of p-JNK and c-jun were also detected by Western blot.Results Subsequent tumor size and weight in DS or DS/Cu-treated animals were （67.71±2.15） mm3,（33.35±7.74） mm3 and （43.35±4.22） mg,（18.05±2.88） mg.One-way ANOVA analysis indicated that the tumor size and weight in DS or DS/Cu-treated animals were reduced significantly relative to tumors in vehicle-treated animals （F =27.579,P =0.000;F =16.369,P =0.000）.Furthermore,multiple comparisons revealed that the DS or DS/Cu-treated animals had significantly reduced tumor size and weight compared with control animals （all P 〈 0.05）.There were significant differences in tumor size and weight between DS or DS/Cu-treated animals （both P 〈 0.05）.Tumor inhibition rates in DS or DS/Cu group were 63.48 % and 80.24 %,respectively.An increase of apoptosis changes in the xenograft tumor cells in DS or DS/Cu treated mice were more significant.Westem blot showed that the p-JNK and c-jun protein expressions in the tumors were improved after the DS or DS/Cu treatment,more obvious in DS/Cu treatment.Conclusion DS/Cu can inhibit the growth of xenografts,and one possible mechanism may involve the regulation of JNK signal pathway.