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| 腺病毒介导的hepaCAM基因对膀胱癌细胞周期的影响 |
| 吕长坤1,2,刘琪3,马菲菲2,罗春丽1*,徐新1 |
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(1.重庆医科大学检验医学院实验诊断学教研室,重庆 400016
2.商丘医学高等专科学校医学检验技术教研室,商丘 476100
3.重庆市涪陵中心医院检验科,重庆 408000
*通信作者) |
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| 摘要: |
| 目的：以腺病毒为载体，研究hepaCAM基因对膀胱癌细胞T24和BIU-87的细胞周期影响。方法：以带有hepaCAM基因的腺病毒载体pAdH5-hepaCAM感染两株细胞，采用RT-QPCR和Western-blot、细胞免疫荧光方法检测感染pAdH5-hepaCAM组，pAdH5空载组和未感染组的hepaCAM mRNA 、蛋白的表达及其细胞定位。并采用流式细胞术检测hepaCAM对两株细胞周期影响，Western-blot方法检测细胞周期蛋白cyclinD1。结果：感染hepaCAM的T24和BIU-87细胞，hepaCAM mRNA和蛋白水平明显升高。流式结果显示hepaCAM阻滞T24与BIU-87细胞于G0/G1期。Western-blot检测实验组cyclinD1明显低于空载和空白组。细胞免疫荧光显示hepaCAM在单个细胞中主要表达于细胞浆中，在细胞之间表达于细胞连接处。结论：hepaCAM能阻滞膀胱癌细胞T24和BIU-87于G0/G1期，并使G1期关键周期蛋白cyclinD1下调。 |
| 关键词: 膀胱肿瘤 hepaCAM 细胞周期 细胞周期蛋白D1 腺病毒 |
| DOI： |
| 投稿时间：2013-03-11修订日期：2013-06-17 |
| 基金项目: |
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| Effect of adenovirus-mediated hepaCAM gene expression on cell cycle of bladder cancer cell lines |
| L Chang-kun1,2,LIU Qi3,MA Fei-fei2,LUO Chun-li1*,XU Xin1 |
(1. Department of Laboratory Diagnosis, College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016,China
2. Department of Laboratory Medicine, Shangqiu Medical College, Shangqiu 476100, Henan, China
3. Department of Clinical Laboratory, Fuling Central Hospital, Chongqing 408000, China
*Corresponding author.) |
| Abstract: |
| Objective: our study was to investigate the effect of hepaCAM gene on cell cycle of bladder cancer cell line T24 and BIU-87. Method: Adenovirus vector with hepaCAM was infected into T24 and BIU-87 cells. The expression of hepaCAM mRNA and protein were measured byRT-QPCR and Western-blot. Immunofluorescence detected the cellular localization of hepaCAM. The distribution of cell cycle was analyzed by FCM. The protein expression of cyclinD1 was measured by Westein-blot. Result: The expression of hepaCAM mRNA and protein was increased in two cell lines infected with pAdH5-hepaCAM. The G0/G1 cycle was increased in infected cells, and the protein expression of cyclinD1 was decreased in same cells. Immunofluorescence revealed that hepaCAM protein localized on cytoplasm in well-spread cells, otherwise it localized on the junction of each cell. Conclusion: T24 and BIU-87 cells that infected pAdH5- hepaCAM was blocked in G0/G1 cycle, and it can decreased the protein expression of cyclinD1. |
| Key words: urinary bladder neoplasms hepaCAM cell cycle cyclinD1 adenovirus |
