抑制PDK对人结肠癌细胞增殖和化疗敏感性影响及其机制的探讨

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目的:探讨抑制丙酮酸脱氢酶激酶(PDK)对结肠癌细胞增殖和化疗敏感性的影响及可能机制。方法:应用MTT法观察PDK抑制剂(DCA)对人结肠癌细胞化疗增敏效应;应用Hoechst33258染色通过激光共聚焦显微镜观察DCA诱导的LoVo细胞凋亡;用流式细胞术观察DCA诱导的细胞凋亡、线粒体膜电位和Ca2+通道开放的情况。结果:抑制PDK(DCA)能明显影响结肠癌细胞增殖,其中90mmol/LDCA刺激SW620、LoVo、HT29、LS174T和293T细胞48h后,活性细胞率分别为(23.90±2.79)%、(5.90±1.31)%、(32.82±4.50)%、(29.72±3.03)%和(84.86±0.50)%。与小剂量DCA(2.5和5mmol/L)联合作用LoVo细胞48h,低剂量5-FU(5mmol/L)的活性细胞率由(66.03±3.95)%分别降至(50.78±2.05)%和(39.8±2.86)%;而低剂量奥沙利铂(0.5mmol/L)的活性细胞率由(71.18±0.62)%,分别降至(58.24±3.27)%和(53.17±2.65)%。30mmol/LDCA作用4种结肠癌细胞48h后,LS174T、SW620、HT29、LoVo和293T凋亡的发生率分别为(22.27±1.75)%、(14.42±2.36)%、(25.55±0.78)%、(13.20±1.58)%和(10.73±1.49)%。30mmol/LDCA作用LoVo细胞48h后,线粒体膜电位值由4.04±1.47下降为2.09±0.86;胞内Ca2+浓度下降了83.6%。结论:抑制PDK能抑制结肠癌细胞增殖活性,增加化疗敏感性。该作用可能是通过线粒体功能异常引起结肠癌细胞凋亡。 Objective: To investigate the effects of pyruvate dehydrogenase kinase (PDK) on the proliferation and chemosensitivity of human colon cancer cells and its possible mechanism. Methods: The effect of PDK inhibitor (DCA) on chemosensitivity of human colon cancer cells was observed by MTT assay. The apoptosis of LoVo cells induced by DCA was observed by laser confocal microscopy with Hoechst33258 staining. The apoptosis of DCs induced by DCA was observed by flow cytometry Death, mitochondrial membrane potential and Ca2 + channel open situation. Results: The inhibition of PDK (DCA) significantly affected the proliferation of colon cancer cells. The rate of active cells of SW620, LoVo, HT29, LS174T and 293T cells stimulated by 90mmol / LDCA for 48h were (23.90 ± 2.79)%, (5.90 ± 1.31) %, (32.82 ± 4.50)%, (29.72 ± 3.03)% and (84.86 ± 0.50)%, respectively. LoVo cells were treated with low doses of DCA (2.5 and 5 mmol / L) for 48 h, and the activity of low doses of 5-FU (5 mmol / L) decreased from (66.03 ± 3.95)% to (50.78 ± 2.05)% and ± 2.86)%, while the activity of low dose oxaliplatin (0.5mmol / L) decreased from (71.18 ± 0.62)% to (58.24 ± 3.27)% and (53.17 ± 2.65)%, respectively. The apoptosis rates of LS174T, SW620, HT29, LoVo and 293T cells after treated with 30 mmol / L LDCA for 48 h were (22.27 ± 1.75)%, (14.42 ± 2.36)%, (25.55 ± 0.78)%, 13.20 ± 1.58)% and (10.73 ± 1.49)% respectively. After LoVo cells were treated with 30mmol / L LDCA for 48h, the mitochondrial membrane potential decreased from 4.04 ± 1.47 to 2.09 ± 0.86, and the intracellular Ca2 + concentration decreased by 83.6%. Conclusion: Inhibition of PDK can inhibit the proliferation of colon cancer cells and increase the chemosensitivity. This effect may be caused by mitochondrial dysfunction of colon cancer cells apoptosis.
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