姜黄素脂质体通过调节miRNA的表达改变乳腺癌细胞对阿霉素的化疗敏感性

Curcumin-Loaded Liposome Alters Chemosensitivity of Breast Cancer Cells to Adriamycin by Regulating the Expression of MicroRNAs

目的初步研究姜黄素脂质体(CUR)在人乳腺癌细胞株MCF-7中对阿霉素(ADR)药物敏感性的影响,并分析其通过调控miRNA途径的作用机制。方法以人乳腺癌细胞株MCF-7为亲本细胞,采用低浓度逐步加量法诱导阿霉素耐药株MCF-7/ADR,利用薄膜法制备姜黄素脂质体,MTT法检测姜黄素脂质体增加MCF-7/ADR耐药株敏感性的性能及与阿霉素的协同增效作用,miRNA和mRNA微阵列分析筛选出差异表达的miRNA和mRNA,联合生物信息学预测软件和mRNA微阵列预测差异表达的miRNA可能的靶基因,应用荧光定量PCR验证部分微阵列结果。结果阿霉素与姜黄素脂质体对MCF-7/ADR的细胞抑制率有协同增效作用。芯片结果显示,与耐药株MCF-7/ADR细胞相比,miRNA和mRNA在亲本MCF-7/ADR细胞和经姜黄素脂质体处理后的MCF-7/ADR细胞中的表达差异明显,共发现67个差异表达的miRNA和323个差异表达的mRNA。根据预测软件和mRNA芯片结果发现20个共同靶基因。qPCR进一步验证hsa-miR-29b-1-5p,hsamiR-29b-3p,hsa-miR-6068,hsa-miR-6790-5p,has-miR-4417和DDIT4,EPAS1,VEGFA,RPS14,DCDC2在3株细胞中的表达与芯片结果基本一致。结论姜黄素脂质体能在一定程度上逆转人乳腺癌对阿霉素的耐药性。通过调控miRNA的表达来影响相关信号通路可能是姜黄素脂质体调节乳腺癌细胞对阿霉素药物敏感性的一条重要分子途径。

OBJECTIVE To investigate the effect of curcumin-loaded liposome on the chemosensitivity of breast cancer cells to adriamycin via regulating miRNA pathway,and to study its possible mechanism.METHODS Breast cancer cell line MCF-7/ADR was established in vitro by gradually increasing the concentration of adriamycin from the parent cell line MCF-7.The effect of curcumin-loaded liposome altering chemosensitivity of ADR-resistant MCF-7 human breast cancer（MCF-7/ADR）and the synergistic effect index（CI）of ADR and CUR were observed by MTT assay.Differentially expressed miRNAs and mRNAs were detected using miRNA and mRNA microarray.Candidate genes of dysregulated miRNAs were identified by prediction algorithms based on gene expression profiles.RESULTS The combination use of ADR and curcumin-loaded liposome markedly increased inhibition ratio of MCF-7/ADR cells.The array results showed significant differential expression of miRNA and mRNA among MCF-7/S,MCF-7/ADR and curcumin-treated MCF-7/ADR.There were 67 miRNAs and 323 mRNA were differentially expressed in three cell lines.20 target genes of each dysregulated miRNA not only predicted by prediction software but also differentially expressed in the microarray were discovered.The expression status of hsa-miR-29 b-1-5 p,hsa-miR-29 b-3 p,hsa-miR-6068,hsa-miR-6790-5 p,has-miR-4417 and DDIT4,EPAS1,VEGFA,RPS14,DCDC2 was validated using qPCR.The results were basically consistent with the analysis results of miRNA microarray.CONCLUSION Curcumin-loaded liposome can rescue part of adriamycin resistance in breast cancer.An altered miRNA expression pattern is involved in acquiring resistance to ADR and miRNA signaling pathways maybe an important molecular pathway through which curcumin-loaded liposome can change the resistance to ADR in breast cancer MCF-7 cells.