腺病毒载体介导的PML生长抑制因子用于胆囊癌基因治疗的实验研究

目的 探讨腺病毒介导的PML基因对胆囊癌细胞（GBC-SD）的生长抑制作用。方法 用原位杂交和免疫组织化学法分别检测PMI，重组腺病毒的转导效率、GBCSD中PMI，mRNA和蛋白的表达量及表达时间，MTT法检测PML基因转染后GBC-SD的生长状况，观察转染PML基因后GBC-SD的致瘤能力的变化。结果 腺病毒滴度为100MOI时被感染的胆囊癌细胞可达100％，Ad-PML感染的GBC-SD能高效表达PML-mRNA和PML蛋白，表达时间可持续2周以上，PML对GBC-SD体外生长具有明显抑制作用，Ad-PML感染的GBC-SD不能在裸鼠体内形成肿瘤。结论 腺病毒介导的PML基因能在GBC-SD细胞中稳定、高效表达，并能显著抑制GBC-SD细胞的体外生长及其在裸鼠体内的致瘤性，PML有可能用于胆囊癌的基因治疗。

Adenovirus-mediated PML growth suppressor in gene therapy of gallbladder carcinoma

Objective To investigate the inhibitory effects of adenovirus-mediated PML growth suppressor on growth of gallbladder carcinoma cells.Methods Transduction efficiency of PML gene was tested by in situ hybridization(ISH).PML mRNA and PML protein expression after Ad-PML infection was examined by means of ISH and imunohistochemistry.Gallbladder carcinoma cells(GBC-SD) were placed in 24-well tissue culture plates,and infection was carried out with Ad-PML or Ad-control.The material was analyzed by MTT assay. GBC-SD cells infected by wither Ad-PML or Ad-control and positive control were resuspended,and the cell suspension was subcutaneously injected into the nude mice.Tumor formation was evaluated at the end of a 1-month period after the injection,and the tumor volume and weight were determined.Results The maximal transduction efficiency of PML was obtained at a concentration of 100 MOI.The Ad-PML-infected cells expressed a high level of PML mRNA and protein.A detectable level of PML was found on day 15 after viral infection.The growth rate of the Ad-PML-infected GBC-SD cells was significantly inhibited,and the Ad-PML-treated GBC-SD cells failed to form tumors in nude mice.Conclusions PML gene is highly and stably expressed in Ad-PML-infected GBC-SD cells.PML is able to suppress growth in vitro and tumorigenicity of human gallbladder carcinoma cells in nude mice.Ad-PML may be a potential candidate for gene therapy of human gallbladder carcinoma.