肾细胞癌GRC-1细胞系在细胞周期中Wnt-5A基因的不同表达
Different Wnt-5A gene expressions in the renal cell carcinoma GRC-1 cell line during the cell cycle.
作者信息
Zhuang L, Guo H, Xin D, Zhang Z, Li H, Yuan X, Tang D, Ding Y, Liu L, Guo Y
机构信息
Urological Department, First Hospital, Institute of Urology, Beijing Medical University, Beijing 100034, China.
出版信息
Chin Med J (Engl). 2000 Apr;113(4):306-9.
OBJECTIVE
To investigate the gene expression at transcription level of growth factor Wnt-5A in different phase during the cell cycle.
METHODS
We synchronized the renal cell carcinoma GRC-1 cell line by double thymidine blocks and high-pressure N2O gae methods and amplified Wnt-5A cDNAs from different phase using Semi-quantitative RT-PCR (reverse transcriptase polymerase chain reaction). The PCR products were electrophoresized on the agrose gel and detected by Gel Doc 1000 computer controlled system integrating the volumes of each band, representing the intensities of all pixels in a defined band.
RESULTS
The different mRNA expressions of growth factor Wnt-5A was detected in RCC GRC-1 cell line. In S phase, the highest level of Wnt-5A transcript was observed, and in G1 and M phase, medial and lowest, respectively. The differences between S and M stages were statistically significant (P < 0.05).
CONCLUSION
Growth factor Wnt-5A has the potential effect on tumorigenesis. It contributes to all phases during cell cycle but in S phase especially.
目的
研究生长因子Wnt-5A在细胞周期不同阶段转录水平的基因表达情况。
方法
采用双胸腺嘧啶核苷阻断法和高压N₂O气体法使肾癌细胞系GRC-1细胞同步化，运用半定量逆转录聚合酶链反应（RT-PCR）从不同阶段扩增Wnt-5A cDNA。PCR产物在琼脂糖凝胶上进行电泳，通过Gel Doc 1000计算机控制系统检测，该系统可整合各条带的体积，代表特定条带中所有像素的强度。
结果
在肾癌细胞系GRC-1中检测到生长因子Wnt-5A不同的mRNA表达。在S期观察到Wnt-5A转录本水平最高，在G1期和M期分别为中等水平和最低水平。S期和M期之间的差异具有统计学意义（P < 0.05）。
结论
生长因子Wnt-5A对肿瘤发生具有潜在影响。它在细胞周期的各个阶段都有作用，但在S期尤为明显。
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