摘要
目的研究表观遗传药物组蛋白去乙酰化抑制剂曲古霉素A(TSA)对miR-34家族(miR-34a/b/c)在肝癌细胞中表达的影响及其抑制肝癌细胞糖酵解的分子机制。方法在肝癌细胞系中分别转染miR-34模拟物或相应阴性对照,用real-time PCR法检测miR-34a/b/c的表达以及糖酵解途径关键酶的表达;在肝癌细胞中分别转染miR-34b模拟物、相应阴性对照,或用TSA、DMSO处理肝癌细胞,用乳酸检测试剂盒、葡萄糖检测试剂盒分析miR-34b及TSA对肝癌细胞Hep G2、PLC/PRF/5糖酵解途径的影响;设计拯救实验在Hep G2细胞中研究TSA、miR-34b与肝癌细胞糖酵解调控的关系。结果 TSA可以诱导Hep G2、PLC/PRF/5肝癌细胞内源性miR-34b的表达上调(P<0.01);过表达miR-34b可以抑制糖酵解关键酶LDH-A的表达(P<0.05);miR-34 b可以抑制含有LDH-A 3'非翻译区的报告基因活性;敲低miR-34 b的表达可以降低TSA对肝癌细胞Hep G2糖酵解途径的抑制作用。结论 TSA通过诱导miR-34 b表达上调发挥抑制肝癌细胞的代谢转换。
Objective To investigate the effect of TSA on the expression of miR-34 family（ miR-34 a / b / c） and to explore the molecular mechanism of TSA for inhibiting glycolysis in human hepatocellular carcinoma（ HCC）cells. Methods Real-time PCR analysis was conducted to evaluated the expression levels of miR-34 family in HCC cells with TSA treatment. The expression of specific genes involved in the regulation of glycolysis was determined by using Real-time PCR in HCC cells with miR-34 b overexpression or TSA treatment. The effect of miR-34 b or TSA treatment on the glycolysis in HCC cells was investigated by detecting the cellular lactate production and glucose consumption. Rescue assay was performed to clarify the correlation between TSA,miR-34 b,and the regulation of glycolysis in HepG2 cells. Results The expression of miR-34 b was increased in HepG2 and PLC /PRF /5 HCC cells with TSA treatment（ P 〈0. 01）. Enforced expression of miR-34 b led a to reduced level of LDH-A in these two cells（ P〈0. 05）. Rescue assay revealed that inhibition of miR-34 b to prevent the TSA induction resulted in suppressed glycolysis in HepG2 HCC cells. Conclusions TSA inhibits the metabolic shift inHCC cells through inducing the expression of miR-34 b.
出处
《基础医学与临床》
CSCD
2016年第6期772-776,共5页
Basic and Clinical Medicine
基金
国家自然科学基金(81570780
81372578)
