肝细胞癌中DCLK1转录调控的机制研究

Mechanism of DCLK1 transcriptional regulation in HCC

目的:研究双皮质素样激酶1(doublecortin-like kinase 1,DCLK1)在肝癌中发生转录上调的机制,为肝癌的防治提供新思路。方法:实时定量PCR检测表皮生长因子(epidermal growth factor,EGF)是否通过表皮生长因子受体(epidermal growth factor receptor,EGFR)上调肝癌细胞中DCLK1 mRNA的表达,Western blot检测EGF是否通过EGFR上调肝癌细胞中DCLK1蛋白的表达,免疫组织化学染色检测EGFR和DCLK1在39例人肝癌组织中的表达情况。结果:EGF孵育肝癌细胞HepG2和Huh-7后,DCLK1 mRNA和蛋白的表达明显上调(P<0.05,P<0.01),而基因敲减EGFR可以显著抑制EGF诱导的DCLK1 mRNA和蛋白的上调(P<0.01)。在人肝癌标本中,EGFR和DCLK1表达呈显著的正相关(r=0.6696)。结论:EGFR参与了肝癌组织中DCLK1的转录调控。

AIM:To investigate the molecular mechanism through which DKK1 is transcriptionally regulated in HCC(hepatocellular carcinoma)cells.METHODS:Real time PCR was used to explore whether EGFR was involved in regulating DCLK1 mRNA expression in HCC cells;Western blot assay was used to examine whether EGFR-mediated the up-regulation of DCLK1 protein in HCC cells;Immunohistochemical(IHC)analyses were used to examine the protein expression of EGFR and DCLK1 in 39 human HCC tumor specimens.RESULTS:EGF promoted the expression of DCLK1 mRNA and protein in HepG2 and Huh-7 cells(P<0.05,P<0.01),while knockdown of EGFR with two specific siRNA could reverse EGF-induced the up-regulation of DCLK1 mRNA and protein(P<0.01).IHC analyses revealed that the amount of EGFR correlated significantly with that of DCLK1(r=0.6696).CONCLUSION:EGFR promoted DCLK1 transcription in HCC.