遗传印记基因PEG 10转基因小鼠的建立及其皮下移植瘤生长和转移的变化

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、Western blot鉴定后,皮下注射H22细胞,连续测量肿瘤体积.12 d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枪测到目的 基因和蛋白的表达.转皋因小鼠皮卜瘤的体积(4.08、4.23 cm3)及质量(6.89、6.48 g)均显著高于野生昔小鼠(1.61 cm3及1.63 g,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卜移植瘤的生长、侵袭和转移.

Establishment of PEGI0 transgenic mouse and effects of PEG10 on growth, metastasis of transplanted tumor in mice

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-PEG 10, which contained mouse albumin promoter, structural gene of PEG 10, 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 gcnomic DNA in tail tissue, then PEG 10 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 PEG 10 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 transgenie mice invaded the surrounding tissues and showed liver metastasis, PEG I0 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.