CYB5D2对乳腺癌皮下移植瘤模型大鼠肿瘤生长的影响  

Effect of CYB5D2 on tumor growth in subcutaneous transplantation mouse model of breast cancer

作　　者：贠浩 周锦纹 张锋 肖艾依 YUN Hao;ZHOU Jin-wen;ZHANG Feng;XIAO Ai-yi(Department of Breast Surgery,Shanxi Provincial Cancer Hospital,Xi'an 710061,China)

机构地区：[1]陕西省肿瘤医院乳腺科,陕西西安710061

出　　处：《生物技术》2022年第2期177-181,共5页Biotechnology

摘　　要：[目的]研究CYB5D2对乳腺癌皮下移植瘤模型大鼠肿瘤生长的影响。[方法]通过乳腺癌MCF-7细胞系建立乳腺癌裸鼠皮下移植瘤动物模型。将含有CYB5D2表达的质粒稳定转染MCF-7细胞,多点注射,将稳定转染的细胞注入裸鼠皮下,作为观察组,同时设置shRNA-CYB5D2阴性质粒为阴性对照组,PBS为空白对照组。测量各组大鼠肿瘤体积及质量;裸鼠转移瘤模型建立成功后处死小鼠,qRT-CPR法检测肿瘤组织内CYB5D2 mRNA表达水平;Western Blot法检测各组裸鼠转移瘤组织内上皮细胞间质转化(EMT)标志表白表达情况,酶联免疫吸附法检测裸鼠转移瘤细胞上清液中MMP-2及MMP-9水平。[结果]观察组转移瘤平均体积及质量显著低于阴性组与空白组(P<0.05),阴性组与空白组裸鼠转移瘤体积及质量无明显差异(P>0.05)。提取试验小鼠肿瘤组织总RNA,qRT-CPR法检测提示观察组肿瘤组织内CYB5D2 mRNA表达量显著高于阴性组及空白对照组(P<0.05),而阴性组与空白组间差异无显著性差异(P>0.05);Western Blot法检测结果显示,观察组转移瘤组织内E-cadherin、β-catenin蛋白表达水平显著高于与阴性组及空白组(P<0.05),而N-cadherin及Snail蛋白表达显著低于与阴性组及空白组(P<0.05)。阴性组与空白组转移瘤组织内E-cadherin、β-catenin、N-cadherin及Snail蛋白表达无显著性差异(P>0.05);观察组裸鼠转移瘤细胞上清液中MMP-2及MMP-9水平显著低于与阴性组及空白组(P<0.05),而阴性组与对照组MMP-2及MMP-9水平无显著性差异(P>0.05)。[结论]上调CYB5D2基因可有效抑制乳腺癌皮下移植模型大鼠瘤体生长,推测上调CYB5D2基因可能通过下调N-cadherin、Snail、MMP2及MMP9,上调E-cadherin、β-catenin,逆转EMT,发挥抗肿瘤功效。[Objective] To investigate the effect of CYB5 D2 on tumor growth in subcutaneous transplantation mouse model of human breast cancer.[Method] Human breast cancer cells MCF-7 cells were transplanted subcutaneously to mice to generate tumors.Rats receiving multi-point injection of MCF-7 cells containing CYB5 D2 were set as observation group;rats receiving multi-point injection of MCF-7 cells containing shRNA-CYB5 D2 were set as negative control group,and those receiving PBS injection were set as blank control group.Tumor volume and mass were measured.After model establishment,qRT-CPR was used to detect the mRNA expression level of CYB5 D2 in tumor tissues;Western Blot was used to detect the expression of epithelial-mesenchymal transition(EMT) related markers in metastatic tumor tissues;meantime,enzyme-linked immunosorbent assay was used to detect the expression of MMP-2 and MMP-9 in the serum of metastatic tumor cells in nude mice.[Result]The average volume and mass of metastatic tumor in observation group were significantly lower than those in negative control group and blank control group(P<0.05),and no significant difference was found between negative control group and blank control group(P>0.05).The qRT-CPR method showed that the mRNA expression of CYB5 D2 in tumor tissues of observation group was significantly higher than that of negative control group and blank control group(P<0.05),and the expression showed no significant difference between negative control group and blank control group(P>0.05).Western Blot showed that the expression levels of E-cadherin and β-catenin protein in metastatic tumor tissues of observation group were significantly higher than those of negative control group and blank control group(P<0.05),and the expression levels of N-cadherin and Snail protein were significantly lower than those of negative control group and blank control group(P<0.05).No statistical difference was found in the expression of E-cadherin,β-catenin,N-cadherin and Snail protein between negative control group and b

关 键 词：CYB5D2 乳腺癌 皮下移植瘤大鼠模型 肿瘤生长 

分 类 号：R737.9[医药卫生—肿瘤]