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目的探究单羧酸转运蛋白1(MCT1)在增强乳腺癌细胞对3-溴丙酮酸(3-Br PA)敏感性中的作用,为乳腺癌治疗提供新思路。方法 MTT法检测3-Br PA对乳腺癌细胞的增殖抑制作用,溴化丙啶单染法流式细胞术检测细胞凋亡,ELISA试剂盒检测细胞内己糖激酶Ⅱ、乳酸脱氢酶、乳酸、三磷酸腺苷水平,Western blot检测MCT1蛋白的表达,瞬时转染c DNA上调MCT1的表达后,检测3-Br PA对MDA-MB-231细胞的增殖和三磷酸腺苷水平的影响。结果 3-Br PA对MDA-MB-231细胞增殖和凋亡的影响不明显,200μmol/L作用24 h后增殖抑制率和凋亡率仅为8.72%和7.8%;但200μmol/L 3-Br PA作用MCF-7细胞24 h后其增殖抑制率和凋亡率为84.6%和82.3%。MDA-MB-231细胞的MCT1过表达后,200μmol/L 3-Br PA对细胞的增殖抑制率为72.44%,明显高于对照组(P<0.05);25、50、100、200μmol/L 3-Br PA作用细胞6 h后,细胞内三磷酸腺苷水平和对照组相比分别为96.98%、88.44%、43.3%、27.56%。结论 MCT1能增强乳腺癌细胞对3-Br PA的敏感性,其机制可能是将3-Br PA转运到细胞内,从而抑制细胞糖酵解来发挥抗肿瘤作用。
Objective To investigate the role of MCT1 in enhancing the sensitivity of breast cancer cells to 3-bromopyruvate (3-Br PA) and to provide a new idea for the treatment of breast cancer. Methods The inhibitory effect of 3-Br PA on the proliferation of breast cancer cells was detected by MTT assay. Apoptosis was detected by flow cytometry with propidium iodide staining. The levels of hexokinase Ⅱ, lactate dehydrogenase, , The level of adenosine triphosphate (ATP), the expression of MCT1 protein by Western blot, and the effect of 3-Br PA on the proliferation of MDA-MB-231 cells and the level of adenosine triphosphate (ATP). Results The proliferation and apoptosis of MDA-MB-231 cells were not significantly affected by 3-Br PA. The proliferation and apoptosis rates of MDA-MB-231 cells treated with 200μmol / L were only 8.72% and 7.8% The proliferation and apoptosis rates of PA-treated MCF-7 cells after 24 h were 84.6% and 82.3%, respectively. After MCT1 was over-expressed in MDA-MB-231 cells, the inhibition rate of 200μmol / L 3-Br PA was 72.44%, which was significantly higher than that of the control group (P <0.05); 25,50,100,200μmol / L After 6 h, the levels of adenosine triphosphate (ATP) in cells were 96.98%, 88.44%, 43.3% and 27.56% compared with the control group respectively. Conclusion MCT1 can enhance the sensitivity of breast cancer cells to 3-Br PA. Its mechanism may be to transport 3-Br PA into cells and inhibit the cell’s glycolysis to exert anti-tumor effect.