摘要
目的研究 p16基因和顺铂联合应用对胆管癌细胞的作用.方法将重组体腺病毒 p16(Ad-p16)和顺铂联合作用于人胆管癌细胞 QBC939,对 p16基因的表达、细胞的生长抑制及机制进行分析.结果用 Ad-LacZ 进行重组体腺病毒转导效率的检测,发现当MOI 为100以上时,重组体腺病毒可使90%以上的培养的人胆管癌 QBC939细胞被转导.用 RT-PCR 方法检测,在胆管癌QBC939细胞系中 p16呈低表达.重组体腺病毒能介导外源基因 p16在胆管癌 QBC939细胞系中高效表达,重组体腺病毒介导的 p16在 QBC939细胞中表达,能抑制 QBC939细胞的生长和集落形成.其与顺铂联合应用对 QBC939细胞的生长抑制具有明显作用.并显著地抑制该肿瘤细胞的克隆形成能力.流式细胞计数证实 p16能诱导 PBC939细胞发生凋亡并导致其发生 G_1期阻滞,顺铂能诱导 QBC939细胞发生凋亡并导致细胞发生明显的 G_2期阻滞.结论 p16基因能够增加 QBC939细胞对顺铂的敏感性.
AIM To evaluated the inhibitory effect of p16 with cisplatin (CDDP) on human cholangiocarcinoma cell line. METHODS The effects of recombinant adenoviruses (Ad- p16) with CDDP on the growth of QBC939 cell in vitro. RESULTS The QBC939 cell line transducted with recombinant adenoviruses LacZ (Ad-LacZ) at MOI of 100 or greater exhibited more than 90% transduction rate.p16 expression was low in QBC939 cells but high in QBC939 cell lines infected with Ad-p16.The growth rates of the Ad-p16-infected QBC939 cells(MOI of 100) were inhibited by 35.1% (8 day),the growth rates of the QBC939 cells by 56.0% or 67.8% with cisplatin (CDDP,0.5 or 1.0mg/L,8 day) and the growth rates of the Ad-p16- infected QBC939 cells (MOI of 50) with CDDP (0.5 mg/L) by 77.7% (8 day),compared with that of the Ad-LacZ- infected cells.Colony formation in these Ad-p16-infected with CDDP cells greatly decreased versus Ad-LacZ infected cells.The Ad-p16 transfected cells induced QBC939 cell apoptosis and G^1 blockade which conformed by the flow cytometry.The suppression effects mediated by CDDP in tumor cell results mainly from apoptosis and G^2 arrest. CONCLUSION p16 gene can increase the chemosen- sitivity of cisplatin on human cholangiocarcinoma cells.
出处
《世界华人消化杂志》
CAS
2000年第6期641-645,共5页
World Chinese Journal of Digestology
基金
863高科学技术发展资助项目
No.Z20-01-02
关键词
胆管癌
P16基因
腺病毒
顺铂
基因治疗
凋亡
cholangiocarcinoma
p16 gene
adenovirus
cisplatin
gene therapy
apoptosis