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沉默PCAF基因抑制人肾上腺皮质癌裸鼠移植瘤生长及血管生成的实验研究[J]. 肿瘤防治研究, 2015, 42(05): 454-458. DOI: 10.3971/j.issn.1000-8578.2015.05.007
引用本文: 沉默PCAF基因抑制人肾上腺皮质癌裸鼠移植瘤生长及血管生成的实验研究[J]. 肿瘤防治研究, 2015, 42(05): 454-458. DOI: 10.3971/j.issn.1000-8578.2015.05.007
Inhibitory Effect of Silencing PCAF Gene on Growth and Angiogenesis of Human Adrenocortical Carcinoma Xenograft in Nude Mice[J]. Cancer Research on Prevention and Treatment, 2015, 42(05): 454-458. DOI: 10.3971/j.issn.1000-8578.2015.05.007
Citation: Inhibitory Effect of Silencing PCAF Gene on Growth and Angiogenesis of Human Adrenocortical Carcinoma Xenograft in Nude Mice[J]. Cancer Research on Prevention and Treatment, 2015, 42(05): 454-458. DOI: 10.3971/j.issn.1000-8578.2015.05.007

沉默PCAF基因抑制人肾上腺皮质癌裸鼠移植瘤生长及血管生成的实验研究

Inhibitory Effect of Silencing PCAF Gene on Growth and Angiogenesis of Human Adrenocortical Carcinoma Xenograft in Nude Mice

  • 摘要: 目的 探讨靶向沉默肾上腺皮质癌SW13细胞PCAF基因表达对裸鼠移植瘤生长以及血管生成的抑制作用及其机制。方法 pLenti6.3-MiRNA-PCAF重组慢病毒及pLenti6.3-MiRNA-NC重组慢病毒分别感染SW13细胞,将感染后的两组细胞及未处理的SW13细胞分别接种于15只裸鼠腋下,构建裸鼠移植瘤模型,分为实验组、阴性对照组、空白对照组,并观察各组肿瘤生长情况。Western blot、荧光定量PCR检测移植瘤组织PCAF蛋白、mRNA表达水平,HE染色法观察各组移植瘤组织形态学变化,免疫组织化学法检测VEGF表达及微血管密度。结果 实验组小鼠的肿瘤体积较阴性对照组和空白对照组显著缩小,肿瘤抑制率分别为49.2%和52.9%;PCAF蛋白相对表达水平较阴性对照组下降58.4%,mRNA相对表达水平较阴性对照组下降74.3%;实验组与阴性对照组和空白对照组相比,瘤内VEGF蛋白表达明显下调;实验组微血管密度(19.4±4.2)与阴性对照组(42.7±6.5)和空白对照组(51.3±8.6)相比明显下降。结论 慢病毒介导的pLenti6.3-MiRNAPCAF能有效抑制肾上腺皮质癌细胞裸鼠移植瘤的生长及血管生成。

     

    Abstract: Objective To investigate the inhibitory effect of silencing PCAF gene in adrenocortical carcinoma SW13 cells on tumor growth and angiogenesis, and related mechanism. Methods pLenti6.3-MiRNA-PCAF recombinant lentivirus, and plenti6.3-MiRNA-NC recombinant lentivirus were used to infect the SW13 cells respectively. The infected and untreated cells were inoculated into 15 nude mice's oxter. Fifteen nude mice were randomized into experiment group, negative control group and blank control group. The infected SW13 cells and non-infected SW13 cells were respectively incubated into nude mice to induce the xenografts model. The tumor growth was observed. Western blot and real-time quantitive fluorescent PCR were used to test the protein and mRNA expression of PCAF in the xenografts. HE staining was applied to observe the morphologic changes of transplanted tumor. Immunohistochemical staining was used to detect the positive expression of VEGF and the microvessel density(MVD). Results The average tumor volume in the experiment group was significantly less than those in the negative control and blank control groups with an inhibitory rate of 49.2% and 52.9% respectively. Western blot and qPCR showed that the protein and mRNA expression of PCAF was decreased by 58.4% and 74.3% in the experiment group, compared with the negative control group. The expression of VEGF protein was significantly lower in the experiment group than those in the negative control and blank control groups. MVD was significantly lower in the experiment group(19.4±4.2) than those in the negative control(42.7±6.5) and blank control groups(51.3±8.6). Conclusion pLenti6.3- MiRNA-PCAF mediated by lentiviral vector could effectively inhibit the growth and angiogenesis of adrenocortical carcinoma xenograft in nude mice.

     

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