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遗传印记基因PEG10转基因小鼠的建立及其皮下移植瘤生长和转移的变化 被引量:1

Establishment of PEGIO transgenic mouse and effects of PEG10 on growth, metastasis of transplanted tumor in mice
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摘要 目的 建立PEG10转基因小鼠模型,研究PEG10对小鼠皮下移植瘤的生长和转移的影响。方法PCR阳性转基因鼠经RT—PCR、Westernblot鉴定后,皮下注射H22细胞,连续测量肿瘤体积。12d后取肿瘤和肝脏组织进行HE染色,肝脏组织进行SP染色,检测PEG10蛋白的表达。计量资料采用独立样本的f检验。结果阳性首建鼠的肝脏中检测到目的基因和蛋白的表达。转基因小鼠皮下瘤的体积(4.08、4.23cm3)及质量(6.89、6.48g)均显著高于野生型小鼠(1.61cm。及1.63g,P〈0.05),均向周围组织侵袭并出现肝转移,肝脏中检测到PEG10蛋白;野生型小鼠的肿瘤组织有包膜,未出现肝转移。结论构建的PEG10转基因小鼠模型可促进皮下移植瘤的生长、侵袭和转移。 Objective To establish PEG10 transgenic mice model and study the effect of PEG10 transgene on tumor growth and metastasis in mice. Methods The linearized expression element of pALB- PEG10, which contained mouse albumin promoter, structural gene of PEG10, and polyaenylation signal sequence, was microinjected into 3741 KM mouse fertilized ova. The manipulated embryos were then trans- planted into the oviducts of 94 pseudopregnant recipient mice. All the newborn mice were screened by PCR to detect genomic DNA in tail tissue, then PEG10 mRNA and protein expression were detected by RT-PCR and western blot, respectively in the positive mice. Hepatoma cell H22 was subcutaneously inoculated into the right armpit of wild type mice and No. 17, No.33 transgenic mice. Tumor size was measured every week. Mice were sacrificed on day 12 and then the tumors were exercised and weighted. Tumors and livers were fixed in formaldehyde and sectioned. The sections were stained with hematoxylin/eosin and examined under microscope. The expression of PEG10 protein was detected with immunohistochemistry method. Results Among the 43 off-springs, 3 were positive for tail tissue PEG10 gene examination, PEG10 was successfully expressed in the liver of the randomly selected transgenic mouse. H22 tumor grew faster in all the transgenic mice than in wild type mice. The average size and weight of tumors between the transgenic mice and wild type mice were significantly different (P 〈 0.05). Most tumors in the transgenic mice invaded the surrounding tissues and showed liver metastasis, PEGIO protein was expressed in liver. In contrast, nearly all the tumors in wild type mice were capsulized and PEG10 was not expressed in liver. Conclusion Our results showed that the PEG10 gene could be expressed in the liver of the transgenic mice. PEG10 promotes growth, invasion, and metastasis of transplanted H22 tumors in mice.
出处 《中华肝脏病杂志》 CAS CSCD 北大核心 2009年第6期455-458,共4页 Chinese Journal of Hepatology
基金 国家自然科学基金(30471983,30872237) 国家重点基础研究发展计划(973计划,2007CB512900)
关键词 肝肿瘤 人PEGIO基因 转基因小鼠 制备和鉴定 皮下移植瘤模型 Liver neoplasms Human PEG10 gene Transgenic mice Production and characterization Subcutaneous xenograft tumor model
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