莪术醇对乏氧微环境中人乳腺癌细胞株MCF-7生长转移机制的研究  

Research on Growth and Metastatic Mechanism of Curcumol on Human Breast Cancer Cell Line MCF-7 in Hypoxic Microenvironment

作　　者：王丽莎[1] 夏欣欣[1] 李萍萍 刘永惠[1] WANG Lisha;XIA Xinxin;LI Pinging;LIU Yonghui(The First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,Shaanxi,China;Centre for Translational Medicine,The First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,Shaanxi,China)

机构地区：[1]西安交通大学第一附属医院,陕西西安710061 [2]西安交通大学第一附属医院转化医学中心,陕西西安710061

出　　处：《中华中医药学刊》2024年第12期164-170,I0058-I0060,共10页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81702631);国家中医药管理局全国名老中医药专家传承工作室建设项目(国中医药人教函〔2022〕75号);陕西省中医药管理局基金项目(je104)。

摘　　要：目的以乳腺癌为研究对象,研究乏氧微环境下莪术醇对人乳腺癌细胞株MCF-7增殖活性的影响,并进一步通过促肿瘤生长转移因子HIF-1α、CXCR4、CXCL12的mRNA及蛋白水平的表达,来研究莪术醇对乏氧微环境中MCF-7细胞的生长转移机制的影响。方法在常氧条件下,用含有不同浓度的莪术醇用MTT法检测各莪术醇浓度及时间条件下MCF-7细胞的增殖活性。RT-PCR的方法检测乏氧与常氧环境下莪术醇对人乳腺癌细胞株MCF-7中HIF-1α、CXCL12、CXCR4因子mRNA表达的影响。Westen blotting检测乏氧与常氧环境下莪术醇对MCF-7中HIF-1α、CXCL12、CXCR4因子蛋白表达的影响。免疫组化检测乏氧与常氧环境下莪术醇对MCF-7中HIF-1α、CXCL12、CXCR4因子蛋白表达的影响并检测其表达位置。结果莪术醇可明显抑制人乳腺癌细胞株MCF-7细胞的增殖,且药物作用时间越长、浓度越大,抑制率也越高(P<0.05)。HIF-1α的mRNA及蛋白的表达:常氧条件下HIF-1α几乎无表达,乏氧条件下HIF-1α表达明显;且莪术醇处理的乏氧加药组mRNA及蛋白的表达少于乏氧对照组(P<0.05)。免疫组化结果中可见HIF-1α主要表达在MCF-7的胞核中。乏氧条件下较常氧条件下CXCR4和CXCL12的mRNA及蛋白的表达明显增高(P<0.05),对比莪术醇作用后的乏氧加药组与常氧加药组,CXCR4和CXCL12的mRNA及蛋白在乏氧加药组的表达要明显低于常氧加药组(P<0.05)。CXCR4主要表达MCF-7细胞的胞膜及胞质中。CXCL12主要表达在MCF-7细胞的胞膜和胞质中。结论莪术醇对人乳腺癌细胞株MCF-7细胞的增殖有抑制作用,且有浓度及时间梯度依赖关系。HIF-1α在乏氧条件诱导下表达增高,通过HIF-1α增高,可上调CXCR4、CXCL12因子的表达。莪术醇可明显抑制HIF-1α、CXCL12、CXCR4因子表达,且莪术醇在乏氧条件下的抑制能力更强。Objective In this study,the effect of curcumol on the proliferative activity of human breast cancer cell line MCF-7 in a hypoxic microenvironment was investigated with breast cancer as the research object.The effect of curcumol on the growth and metastatic mechanism of MCF-7 cells in a hypoxic microenvironment was further investigated through the expression of tumor growth promoting metastatic factors[hypoxia inducible factor-1α(HIF-1α),C-X-C chemokine receptor type 4(CXCR4)and Chemokine(C-X-C Motif)Ligand 12(CXCL12)]at the level of mRNA and protein.Methods The proliferative activity of MCF-7 cells was measured by MTT assay under normoxic conditions with different concentrations of curcumol,and the effects of curcumol on the mRNA expressions of HIF-1α,CXCL1 and CXCR4 in human breast cancer cell line MCF-7 under hypoxic and normoxic environments were detected by RT-PCR,and the mRNA expressions of HIF-1α,CXCL12 and CXCR4 in human breast cancer cell line MCF-7 were detected by Western blot.The effect of curcumol on protein expressions of HIF-1α,CXCL12 and CXCR4 in MCF-7 cells under hypoxia and normoxia was detected by blotting.Immunohistochemistry method was performed to detect the effects of curcumol on the protein expression of HIF-1α,CXCL12 and CXCR4 in MCF-7 under hypoxia and normoxia.Results Curcumol significantly inhibited the value-added of human breast cancer cell line MCF-7 cells,and the longer the duration of drug action and the higher the concentration,the higher the inhibition rate(P<0.05).The mRNA and protein expression of HIF-1α:There was almost no expression of HIF-1αunder the condition of normoxia,and the expression of HIF-1αwas obvious under the condition of hypoxia,and the expression of mRNA and protein in the hypoxia dosing group treated with curcumol was less than that in the hypoxia control group.The expression was less than that of the anoxic control group(P<0.05).The immunohistochemical results showed that HIF-1αwas mainly expressed in the nucleus of MCF-7.The mRNA and protein expres

关 键 词：莪术醇 乳腺癌 乏氧微环境 HIF-1Α CXCL12/CXCR4 

分 类 号：R285.1[医药卫生—中药学]