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贺 丽, 李梦龙, 申 越, 等.干扰乳酸脱氢酶A表达对ErbB2高表达乳腺癌细胞迁移和侵袭的影响.四川大学学报(医学版),2019,50(1):55-60
干扰乳酸脱氢酶A表达对ErbB2高表达乳腺癌细胞迁移和侵袭的影响
Effect of LDHA Knockdown by siRNA on Migration and Invasion of ErbB2 Overexpressing Breast Cancer Cell Line
  
中文关键词:  乳腺癌 小干扰 RNA 乳酸脱氢酶 A 糖酵解 迁移与侵袭
英文关键词:Breast cancer siRNA Lactate dehydrogenase A Glycolysis Migration and invasion
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中文摘要:
      目的 探讨体外干扰乳酸脱氢酶A(LDHA)表达对人类表皮生长因子受体2(ErbB2)高表达乳腺癌SK-BR-3和MDA-MB-453细胞迁移和侵袭的影响及其分子机制。方法 用siLDHA转染(以转染scramble siRNA为阴性对照)SK-BR-3和MDA-MB-453细胞干扰LDHA的表达;Western blot法检测LDHA蛋白表达水平;Transwell小室检测细胞的迁移和侵袭能力;采用乳酸脱氢酶(LDH)试剂盒测定细胞的LDH活性;采用葡萄糖和乳酸检测试剂盒分别测定培养基中的葡萄糖浓度和乳酸浓度,计算细胞对葡萄糖的摄取量和乳酸生成量。结果 与阴性对照组比较,siLDHA下调SK-BR-3和MDA-MB-453细胞LDHA的蛋白水平( P<0.01);降低细胞的迁移和侵袭能力( P<0.001);下调SK-BR-3细胞的LDH活性,减少两种细胞葡萄糖摄取量和乳酸生成量,差异均有统计学意义( P<0.05)。结论 干扰LDHA表达通过降低糖酵解从而抑制ErbB2高表达乳腺癌细胞的迁移和侵袭。
英文摘要:
      Objective To investigate the effect of small interfering RNA of lactate dehydrogenase A (siLDHA) on migration and invasion of epidermal growth factor receptor 2 (ErbB2) over expressing breast cancer cell line SK-BR-3, MDA-MB-453 and its molecular mechanism. Methods SK-BR-3 and MDA-MB-453 cells were transfected with siLDHA to interfere with the expression of LDHA. The transfection of scramble siRNA was used as negative control. The LDHA protein levels were detected by Western blot ( P<0.01). Cell migration and invasion was detected by Transwell assays. Lactate dehydrogenase (LDH) activity was measured by LDH assay kit. The glucose and lactate concentration in the culture media was determined by glucose and lactate assay kit, respectively, and then glucose uptake and lactate production by the cells were calculated. Results siLDHA downregulated LDHA protein levels in SK-BR-3 and MDA-MB-453 cells ( P<0.01). Compared with negative control group, siLDHA significantly decreased migration and invasion of SK-BR-3 and MDA-MB-453 cells ( P<0.001). siLDHA reduced LDH activity in SK-BR-3 cells, glucose uptake and lactate production in SK-BR-3 and MDA-MB-453 cells, the difference was significant ( P<0.05). Conclusion Knockdown of LDHA by siRNA inhibits the migration and invasion via downregulation of glycolysis in ErbB2 over expressing breast cancer cell line.
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