Department of Clinical Biochemistry, Attikon University Hospital, Athens 12462, Greece.
World J Gastroenterol. 2011 Mar 28;17(12):1614-21. doi: 10.3748/wjg.v17.i12.1614.
To investigate three isoforms of survivin in colorectal adenocarcinomas.
We used the LightCycler Technology (Roche), along with a common forward primer and reverse primers specific for the splice variants and two common hybridization probes labeled with fluorescein and LightCycler-Red fluorophore (LC-Red 640). Real time quantitative polymerase chain reaction (PCR) was performed on cDNAs from 52 tumor specimens from colorectal cancer patients and 10 unrelated normal colorectal tissues. In the patients group, carcinoembryonic antigen (CEA) and CA19-9 tumor markers were also measured immunochemically.
Wild type survivin mRNA isoform was expressed in 48% of the 52 tumor samples, survivin-2b in 38% and survivin-ΔΕx3 in 29%, while no expression was found in normal tissues. The mRNA expression of wild type survivin presented a significant correlation with the expression of the ratio of survivin-2b, survivin-ΔΕx3, survivin-2b/wild type survivin and survivin-ΔΕx3/wild type survivin (P < 0.001). The mRNA expression of wild-survivin and survivin-ΔΕx3 was related with tumor size and invasion (P = 0.006 and P < 0.005, respectively). A significant difference was found between survivin-2b and morphologic cancer type. Also, the ratio of survivin-ΔEx3/wild-survivin was significantly associated with prognosis. No association was observed between the three isoforms and grade, metastasis, Dukes stage and gender. The three isoforms were not correlated with CEA and CA19-9.
Survivin isoforms may play a role in cell apoptosis and their quantification could provide information about clinical management of patients suffering from colorectal cancer.
研究结直肠腺癌中三种生存素异构体。
我们使用 LightCycler 技术(罗氏),使用一个共同的正向引物和针对剪接变异体的反向引物,以及两个共同的标记有荧光素和 LightCycler-Red 荧光染料(LC-Red 640)的杂交探针。对 52 例结直肠癌患者肿瘤标本和 10 例无关的正常结直肠组织的 cDNA 进行实时定量聚合酶链反应(PCR)。在患者组中,还通过免疫化学方法测量癌胚抗原(CEA)和 CA19-9 肿瘤标志物。
在 52 个肿瘤样本中,野生型生存素 mRNA 异构体在 48%的样本中表达,生存素-2b 在 38%的样本中表达,生存素-ΔΕx3 在 29%的样本中表达,而在正常组织中未发现表达。野生型生存素的 mRNA 表达与生存素-2b、生存素-ΔΕx3、生存素-2b/野生型生存素和生存素-ΔΕx3/野生型生存素的表达呈显著相关(P < 0.001)。野生型生存素和生存素-ΔΕx3 的 mRNA 表达与肿瘤大小和侵袭有关(P = 0.006 和 P < 0.005)。生存素-2b 与形态学癌型之间存在显著差异。此外,生存素-ΔEx3/野生型生存素的比值与预后显著相关。三种异构体与分级、转移、Dukes 分期和性别无关。三种异构体与 CEA 和 CA19-9 均无相关性。
生存素异构体可能在细胞凋亡中发挥作用,其定量分析可能为结直肠癌患者的临床治疗提供信息。