稳定表达乙肝病毒X蛋白肝癌细胞系的建立及其侵袭性研究

Establishment of CCL13 cell lines containing X protein of HBV and characterization of their invasive potential

目的构建稳定表达乙肝病毒X蛋白的CCL13细胞系，为研究HBx在诱导肝细胞癌侵袭和转移等生物学行为的作用提供基础。方法将已构建的含HBx基因的真核表达载体pcDNA3．1-HBx质粒，采用脂质体介导法转染到人肝CCL13细胞系并检测HBx的稳定表达。通过体内、体外实验，研究目的细胞的侵袭能力。结果应用PCR技术从已转染的CCL13细胞中克隆出HBx基因，检测有无稳定表达HBx基因的两组CCL13细胞生长曲线、集落形成率、划痕愈合和细胞侵袭力，发现转染细胞的生物学行为有很明显的恶性肿瘤细胞表型。结论成功构建了稳定表达X蛋白且具有高侵袭性的CCL13细胞系，发现该细胞系侵袭能力的条件性。

Objective To establish cell lines stably expressing HBx protein and explore their invasive potential. Methods The constructed pcDNA3. 1-HBx plasmid containing the eukaryotic ex pression vector pcDNA3.1 was transfected into the CCL13 cell lines employing gene transfection mediated by lipofectamine. Then the stable expression of HBx gene was determined. Meanwhile, the invasire potential of the cell lines was studied in vivo and in vitro. Results HBx gene was cloned from the transfected CCL13 cell line by PCR. The transfected cells had the phenotype of malignant tumor cells. Conclusions The stable CCL13 cell lines containing HBx gene have been successfully established and their invasive potential depends on the conditional changes.