LAMβ1基因下调VEGF表达抑制人胆管癌裸鼠移植瘤生长

Inhibition of Laminin β1 on growth of biliary duct cancer xenografts in nude mice

目的探讨LAMβ1基因对人胆管癌细胞QBC939裸鼠皮下移植瘤生长的影响。方法构建重组慢病毒LAMβ1过表达载体;将24只裸鼠按随机数字表分为3组,分别为实验组(转染LV-LAMβ1)、阴性对照组(空载慢病毒)以及空白对照组,每组8只。制备密度为1×10^7/ml胆管癌QBC939细胞悬液,取0.2ml分别注入裸鼠皮下,每3d测量瘤体大小,4周后处死裸鼠取出裸鼠皮下瘤体。采用RT-PCR、免疫组化检测瘤组织LAMβ1及VEGF表达。结果成功构建过表达慢病毒LAMβ1载体;与阴性对照组和空白对照组比较,实验组裸鼠皮下移植瘤生长受到抑制,重量与体积明显减小(F体积=18.04,P<0.01;F重量=42.87,P<0.01);实验组裸鼠瘤组织中LAMβ1 mRNA和蛋白水平较阴性对照组及空白对照组增高,转移因子VEGF mRNA和蛋白水平下降(P<0.05)。结论 LAMβ1基因过表达通过下调转移因子VEGF表达抑制胆管癌裸鼠皮下移植瘤的生长。

Objective To explore the effects of LAMB1 gene on growth of xenografts in nude mice. Methods The recombinant chronic virus LAMB1 overexpression vector was constucted. 24 female nude mice were randomly divided into exprimental group(LV-LAMB1), negative control group and blank control group. Colorectal carcinoma QBC939 cell suspension(0.2 ml, 1×10^7/ml) was injected into nude mice. Tumor volume and weight were measured every 3 d. Nude mice were sacrificed 4 weeks later and tumor samples removed. LAMB1 and VEGF expression in tumor samples were detected by RT-PCR and immunohistochemistry. Results Recombinant chronic virus LAMB1 overexpression vector was successfully constructed. Compared with negative control group and blank control group, the growth rate of subcutaneous xenograft significantly decreased in experimental group. The tumor’s size and weight of experimental group were significantly lesser than those of the two others group(Volume, F=18.04, P<0.01;Weight, F=42.87, P<0.01). The expression of LAMB1 were higher than that in the two other groups(P<0.05). The expression of VEGF was higher than that in the experimental groups(P<0.05). Conclusions LV-LAMB1 can inhibit the growth of biliary duct cancer. LAMB1 in QBC939 can inhibit the expression of VEGF, which was related to biliary duct cancer metastasis.