摘要
目的探讨HHV-8 Kl在卡波氏肉瘤发病机制中的作用。方法采集新疆经典型KS患者血清,提取血清HHV-8病毒DNA作为模板,运用PCR扩增HHV-8 ORF Kl,以pcDNA3.1/myc-His(-)A构建真核表达载体,并酶切、测序鉴定,采用不同浓度的重组体及空载体转染不含HHV-8的BJAB细胞,采用Western Blot鉴定HHV-8 K1蛋白的表达,观察细胞形态变化,采用Cell Counting Kit-8试剂盒检测转染细胞的增殖率。结果成功构建了pcDNA3.1/HHV-8 K1真核表达载体,HHV-8 K1在BJAB细胞中的表达量随转染重组体浓度的增大而增加,转染重组体组的细胞增殖率高于同浓度下的空载体组,差异有显著性(P均<0.05)。镜下观察,各组细胞形态无显著性差异,但重组体组细胞数量增多明显,生长旺盛。结论HHV-8 K1可以促进细胞增殖,增殖率的高低与HHV-8 K1的表达量存在一定的关系,K1是HHV-8发挥其致KS机制的一种相关蛋白。
Objective To observe the function of HHV-8 K1 in the pathogenesis of Kaposi＇s sarcoma. Methods Serum of patients with classic kaposi＇s sarcoma from Xing Jiang were collected and HHV-8 DNA was extracted from it. HHV-80RF K1 was amplified by polymerase chain reaction and inserted into eukaryotic expression vector pcDNA3.1/ myc-His（-） A. The positive clones were detected by digestion and sequencing. The positive vectors were transfected to the BJAB cell line in differential concentration. The expression of HHV-8 K1 was assayed by Western blot. Morphology of these cells were observed and the rate of proliferation was detected by CCK-8. Results The eukaryotic expression vectors of human herpesvirus 8 K1 were constructed successfully. The expression quantity of HHV-8 K1 increased with the enhancement of the transfected concentration. The rate of proliferation of cells transfected by the vectors of HHV-8 K1 was higher than that transfected by pcDNA3, 1/ myc-His（-） A. The shape of cells in different team was the same, Conclusion K1 can promote the proliferation of cells without the participation of other protein of HHV-8. The rate of proliferation is associated with the expression of K1 and K1 is related with the pathogenesis of HHV-8.
出处
《中国皮肤性病学杂志》
CAS
北大核心
2009年第11期692-694,744,共4页
The Chinese Journal of Dermatovenereology
基金
国际合作项目(2005DFA30780)
石河子大学科学技术研究发展计划项目
