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目的检测丙酮酸脱氢酶激酶-I(pyruvate dehydrogenase kinase-I,PDK1)在人结肠癌组织和细胞中的表达。探讨抑制PDK1对结肠癌细胞活性的影响。方法应用RT-PCR、荧光定量PCR和免疫荧光法、Western blot检测结肠癌组织、癌旁组织以及4种人结肠癌细胞PDK1基因的表达情况。在PDK1抑制剂(dichloroacetate,DCA)的诱导下,通过MTT法观察结肠癌细胞活性。结果PDK1基因在结肠癌组织中表达较癌旁组织中高;在4种人结肠癌细胞中明显表达。抑制PDK1能抑制结肠癌细胞增殖活性,抑制效应随药物浓度而递增,其中90 mmol/L DCA的活性细胞率分别为(23.90±2.79)%、(5.90±1.31)%、(32.82±4.50)%和(29.72±3.03)%。结论PDK1基因在结肠癌组织和细胞中高表达,在癌旁组织中低表达,抑制PDK1明显抑制结肠癌细胞增殖活性,PDK1可望成为结肠癌有效治疗靶点。
Objective To detect the expression of pyruvate dehydrogenase kinase-I (PDK1) in human colon cancer tissues and cells. To investigate the effect of inhibiting PDK1 on the activity of colon cancer cells. Methods The expression of PDK1 gene in colon cancer tissues, pericarcinoma tissues and human colon cancer cells was detected by RT-PCR, fluorescence quantitative PCR, immunofluorescence and Western blot. Colon cancer cell activity was observed by MTT assay under the induction of PDK1 inhibitor (dichloroacetate, DCA). Results The expression of PDK1 gene in colorectal cancer tissues was higher than that in paracancerous tissues and was significantly expressed in four kinds of human colon cancer cells. The inhibitory effect of PDK1 on the proliferation of human colon cancer cells was inhibited by PDK1. The inhibitory effect of PDK1 was increased with the increase of drug concentration. The rate of active cells of DCA at 90 mmol / L was (23.90 ± 2.79)%, (5.90 ± 1.31)%, (32.82 ± 4.50) And (29.72 ± 3.03)% respectively. Conclusion PDK1 gene is overexpressed in colorectal cancer tissues and cells, and is down-regulated in paracancerous tissues. Inhibition of PDK1 significantly inhibits the proliferation of colon cancer cells. PDK1 may be an effective therapeutic target for colon cancer.