腺病毒介导的p16基因转染对膀胱癌细胞AgNORs表达影响的研究

The study of the effects of adenovirus-mediated p16 gene transfer on the expression of AgNORs of the human bladder carcinoma in vitro

目的 :探讨复制缺陷型腺病毒介导的p16基因转染对人膀胱癌细胞中核仁形成区嗜银蛋白 (AgNORs)表达的影响。 方法 :将 p16重组腺病毒 (Ad p16 )感染p16蛋白表达丧失的人膀胱癌T2 4细胞株 ,银染后 ,用光镜观察肿瘤细胞中AgNORs的数目和大小 ,用图像分析仪定量分析AgNORs表达变化。 结果 :转染Ad p16 48h后T2 4细胞AgNORs的大小、数目比对照组小且少 ;Ad p16转染的T2 4细胞的银染颗粒面积和胞核面积的比值 (I.S % )为4.42± 1.78,Ad LacZ和未转染组的I .S %值分别为 7.5 2± 1.92和 7.95± 1.5 8,实验组和对照组比较有非常显著的差异 (P <0 .0 1) ,对照组间差异不显著 (P >0 .0 5 )。 结论 :AgNORs在Ad p16转染后其含量减少 ,AgNORs的表达与细胞的增生能力密切相关 。

Objectives:To probe the effect of full length p16 gene transfer by replication defective recombinant adenovirus on the expression of nucleolar organizer regions associated proteins(AgNORs) of the human bladder carcinoma. Methods: 48 hours after transfection with Ad p16, the number and size of the AgNORs in tumor celles were observed by silver staining, and then the AgNORs were analyzed with a KL imaging system. Results: The cells infected with Ad p16 showed smaller and less AgNORs than those of the control group over 48 hours after the infection. The I.S% of the cells infected with Ad p16 was 4.42 ± 1.78 (s), the I.S% of the cells infected with Ad LacZ or uninfected was 7.52 ± 1.92 or 7.95 ± 1.58 . The I.S% in group infected with Ad p16 is lower than in control groups with statistic significance( P <0.01). Conclusions: The expression of AgNORs was suppressed after transfection with Ad p16. The expression of AgNORs is related with the proliferation of the bladder tumor cells. AgNORs could be considered as a index to evaluate the ability of proliferation in malignant tumor cells.