Cho Gyu-Seok, Ahn Tae Sung, Jeong Dongjun, Kim Jae-Jun, Kim Chang-Jin, Cho Hyun-Deuk, Park Dong-Kook, Baek Moo-Jun
Department of Surgery, Soonchunhyang University College of Medicine, Cheonan, Korea.
J Korean Surg Soc. 2011 Jun;80(6):404-11. doi: 10.4174/jkss.2011.80.6.404. Epub 2011 Jun 9.
Recently, two alternatively spliced survivin variants, survivin-ΔEx3 and survivin-2B, were identified in a single copy of the survivin gene. It has been reported that the expressions of survivin splice variants significantly correlates with the clinical results in many types of human carcinoma. We investigated the transcription levels of survivin and its splice variants in human colorectal carcinomas, and analyzed correlations between survivin expression levels and clinicopathologic features.
We used Western blot and real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) to analyze the protein and mRNA expression levels of survivin variants in 51 colorectal carcinomas. The quantitative RT-PCR was performed using primer pairs specific for survivin and each of its splice variants, then normalized for the gene that encodes glyceraldehydes-3-phosphate dehydrogenase.
In Western blotting, the protein levels of survivin were higher in the tumor tissue than in normal tissue. The expression of survivin, survivin-2B and survivin-ΔEx3 mRNA was present in 96%, 64.7%, and 82.4% of the samples, respectively. When the pathologic parameters were compared, colorectal cancers of advanced pT stages showed significant decrease in survivin-2B mRNA expression by the quantitative RT-PCR (P < 0.001).
The decreased expression of survivin-2B might be related to tumor progression in colorectal cancers. This finding indicates that alternatively spliced variants of survivin may be involved in refining the functions of survivin during tumor progression.
最近,在单个拷贝的生存素基因中鉴定出两种选择性剪接的生存素变体,即生存素-ΔEx3和生存素-2B。据报道,生存素剪接变体的表达与多种人类癌症的临床结果显著相关。我们研究了人类结直肠癌中生存素及其剪接变体的转录水平,并分析了生存素表达水平与临床病理特征之间的相关性。
我们使用蛋白质印迹法和实时定量逆转录聚合酶链反应(RT-PCR)分析了51例结直肠癌中生存素变体的蛋白质和mRNA表达水平。使用针对生存素及其每个剪接变体的特异性引物对进行定量RT-PCR,然后以编码甘油醛-3-磷酸脱氢酶的基因进行标准化。
在蛋白质印迹法中,肿瘤组织中生存素的蛋白质水平高于正常组织。生存素、生存素-2B和生存素-ΔEx3 mRNA的表达分别在96%、64.7%和82.4%的样本中出现。当比较病理参数时,通过定量RT-PCR,晚期pT分期的结直肠癌中生存素-2B mRNA表达显著降低(P < 0.001)。
生存素-2B表达降低可能与结直肠癌的肿瘤进展有关。这一发现表明,生存素的选择性剪接变体可能参与了肿瘤进展过程中生存素功能的细化。