绵羊肺腺瘤病毒囊膜蛋白引起绵羊绒毛膜滋养层细胞的恶性转化

doi:10.11843/j.issn.0366-6964.2018.05.025

摘要/Abstract

摘要：

为了明确绵羊肺腺瘤病毒（JSRV）囊膜蛋白（Env）的致瘤作用，将重组真核表达质粒pEGFP-C1/exJSRV-env和pEGFP-C1/enJSRV-env分别转染到永生化绵羊绒毛膜滋养层细胞（STCs）中，筛选最佳转染条件并利用软琼脂集落形成试验检测是否引起细胞的恶性转化；用平板克隆试验检测囊膜蛋白的表达对细胞增殖的影响。结果显示：在STCs中转染pEGFP-C1/exJSRV-env质粒后细胞在软琼脂中生长并产生集落，同时表现细胞接触抑制性消失；转染pEGFP-C1/enJSRV-env质粒的STCs则表现为不能在软琼脂中产生集落；平板克隆试验结果经SPSS软件分析，转染pEGFP-C1/exJSRV-env的绵羊绒毛膜滋养层细胞的克隆形成率显著高于转染pEGFP-C1/enJSRV-env细胞、pEGFP-C1空载体的细胞以及未转染的细胞（P<0.01）。exJSRV-Env的表达可以诱导STCs发生恶性转化以及细胞增殖。本试验可以为进一步探讨绵羊肺腺瘤逆转录病毒囊膜蛋白的功能提供理论依据，并为研究绵羊肺腺瘤病的致瘤机制提供新思路。

Abstract:

In order to clarify the tumorigenicity of Jaagsiekte sheep retrovirus (JSRV) envelope protein (Env), the recombinant eukaryotic expression plasmids pEGFP-C1/exJSRV-env and pEGFP-C1/enJSRV-env were transfected into immortalized sheep chorionic trophoblast cells (STCs) respectively. The optimal transfection conditions were screened and the soft agar colony forming assay was used to detect the malignant transformation of the cells, and the effect of the expression of the envelope protein on cell proliferation was detected by plate cloning assay. Results were as follows:Sheep trophoblast cells transfected with pEGFP-C1/exJSRV-env acquired colony forming ability in soft agar and lost contact inhibition, while cells transfected with pEGFP-C1/enJSRV-env can't form colony. The results of plate cloning experiments were analyzed by SPSS software, the clone formation rate of sheep trophoblast cells transfected with pEGFP-C1/exJSRV-env was significantly higher than that of transfected with pEGFP-C1/enJSRV-env, pEGFP-C1 empty carrier and non-transfected (P<0.01). The expression of exJSRV-Env can induce malignant transformation and the proliferation of STCs. This study may provide a theoretical basis for further study on the structure and function of Jaagsiekte sheep retrovirus envelope protein, and provide a new idea for the study of the mechanism of sheep pulmonary adenomatosis.

中图分类号:

S852.659.3

赵娟, 徐斯日古楞, 李慧萍, 刘淑英. 绵羊肺腺瘤病毒囊膜蛋白引起绵羊绒毛膜滋养层细胞的恶性转化[J]. 畜牧兽医学报, 2018, 49(5): 1089-1095.

ZHAO Juan, XU Si-ri-gu-leng, LI Hui-ping, LIU Shu-ying. Malignant Transformation of Sheep Trophoblast Cells Induced by Envelope Protein of Jaagsiekte Sheep Retrovirus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(5): 1089-1095.

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