重组腺病毒HBx体内抑制恶性肿瘤细胞的实验研究

Inhibition of Tumour Cells with Hepatitis B Virus x(HBx) Gene Adenoviral Vector in vivo

目的探讨人乙肝病毒x蛋白(HBx)在体内对肿瘤细胞增殖能力的影响。方法利用连接有增强型绿色荧光蛋白的腺病毒(Ad-eGFP),在体外感染H22肿瘤细胞,观察腺病毒对H22细胞的感染效率;在小鼠腹腔内接种小鼠肝癌H22细胞,建立恶性腹水模型,腹腔注射重组腺病毒HBx(Ad-HBx)、空载腺病毒(Ad-null)或生理盐水(对照组),检测腹水肿瘤细胞内的HBx蛋白表达并观察其对肿瘤细胞增殖能力、腹水形成及腹膜渗透性、红细胞等腹水性状的影响。结果腺病毒体外对H22细胞有高的感染效率。HBx蛋白能在腹水肿瘤细胞中表达,在体内环境中Ad-HBx能改善腹膜的渗透性(P<0.05),抑制小鼠腹水生成(P<0.05),使腹水中的红细胞、癌细胞水平明显减少(P<0.05),流式细胞术分析提示Ad-HBx使腹水中的肿瘤细胞周期有明显G2/M阻滞。结论体内环境中Ad-HBx能在肿瘤细胞内表达HBx蛋白,通过细胞周期阻滞明显抑制肿瘤细胞的增殖能力,有望成为治疗癌性腹水的一种新方法。

Objective To investigate the inhibitory effects of the hepatitis B virus x protein （HBx protein） on tumor in vivo. Methods H22 cells were infected with Ad-eGFP to detect infection efficiency of adenovirus. The BALB/c mouse model of malignant ascites was established by implanting H22 cell intraperitoneally into the animal, Ad-HBx, Ad-null or NS were administered intraperitoneally in BALB/c mice separately to detect HBx expression in H22 cells and effects of HBx on inhibition on proliferation of H22 ceils. Results High efficiency of Ad in infecting H22 cells in vitro was observed. HBx protein was expressed in H22 cells after intraperitoneal injection of Ad-HBx. The effect of Ad-HBx on inhibition of the peritoneal capillary permeability and the ascites accumulation （P〈0.05） ; on reduction of the number of red cells and tumor cells counted in malignant ascites （P〈0.05） . and on inhibition of tumor cell life cycle （the G2/M arrest） in malignant ascites were identified by flow cytometric analysis. Conclusion HBx protein can be expressed in turnout cells and can inhibit the proliferation of tumour cells in vivo, and this might be a new potential treatment for malignant ascites.