Zhu Hong-Xia, Zhang Guo, Wang Yi-Hua, Zhou Cui-Qi, Bai Jin-Feng, Xu Ning-Zhi
Laboratory of Cell and Molecular Biology, Cancer Institute, Chinese Academy of Medical Sciences, Beijing 100021, P.R.China.
Ai Zheng. 2004 Jul;23(7):737-41.
BACKGROUND & OBJECTIVE: Although Wnt pathway plays important role in colorectal carcinogenesis, but the mechanism of this pathway in anti-apoptosis is not clear. This study is to investigate the molecular mechanism of Wnt pathway in anti-apoptosis.
Survivin promoter region was constructed into luciferase reporter system (pGL3-sur1.8kb). The recombinants pGL3-sur1.8kb were cotransfected with pRL-SV40 into HCT116 cells and the activities were detected with Dual-luciferase reporter assay system. Cell apoptosis was analyzed by flow cytometry. Protein level of survivin and beta-catenin was detected by Western Blot.
Survivin could be up-regulated by beta-catenin and down-regulated by TCF4DeltaN in transcriptional level. beta-catenin/TCF4 dependent apoptosis induced by indomethacin could suppress survivin transcription. Overexpression of survivin could partially recover the beta-catenin/TCF4 dependent apoptosis.
Down-regulation of survivin affected by beta-catenin/TCF4 pathway plays an important role in apoptosis induced by NSAIDs indomethacin in HCT116 cells. The beta-catenin/TCF4-survivin pathway may be a potential target in treatment of colon cancer.
尽管Wnt信号通路在结直肠癌发生过程中发挥重要作用，但其抗凋亡机制尚不清楚。本研究旨在探讨Wnt信号通路抗凋亡的分子机制。
将Survivin启动子区域构建到荧光素酶报告基因系统（pGL3-sur1.8kb）中。将重组体pGL3-sur1.8kb与pRL-SV40共转染至HCT116细胞中，并用双荧光素酶报告基因检测系统检测其活性。通过流式细胞术分析细胞凋亡情况。采用蛋白质免疫印迹法检测Survivin和β-连环蛋白的蛋白水平。
在转录水平上，β-连环蛋白可上调Survivin表达，而TCF4DeltaN可下调Survivin表达。吲哚美辛诱导的β-连环蛋白/TCF4依赖性凋亡可抑制Survivin转录。Survivin过表达可部分恢复β-连环蛋白/TCF4依赖性凋亡。
β-连环蛋白/TCF4信号通路影响Survivin表达下调在非甾体抗炎药吲哚美辛诱导的HCT116细胞凋亡中起重要作用。β-连环蛋白/TCF4-Survivin信号通路可能是结肠癌治疗的潜在靶点。
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