目的研究乳腺癌组织中差异性表达的miRNAs以及犀黄丸提取液对体外乳腺癌细胞系候选miRNAs的表达影响。方法采用miRNAs表达谱芯片检测石蜡包埋(FFPE)乳腺癌组织、癌旁组织以及不同分子分型的癌组织之间的miRNAs差异性表达谱,非监督聚类分析软件分析芯片结果;利用实时荧光定量PCR(qPCR)技术检测106例乳腺癌组织、22例癌旁组织以及66例乳腺良性病变组织候选miRNAs的表达水平;犀黄丸提取液干预乳腺癌T-47D细胞及MDA-MB-231细胞,采用qPCR技术检测细胞中候选miRNAs的表达水平。结果 miRNAs表达谱芯片结果显示与癌旁组织相比,miR-130b在乳腺癌组织中呈现高表达(P <0. 05),miR-205在恶性程度更高的Basal-like型癌组织中表达显著低于Luminal型(P <0. 05);qPCR结果显示miR-130b在癌组织和癌旁组织中的表达均显著高于良性病变组织(t=6. 123,P=0. 000;t=-2. 073,P=0. 041),miR-205在癌组织中表达水平显著低于良性病变组织(t=-3. 375,P=0. 001);体外细胞实验显示,犀黄丸提取液干预后,miR-130b在T-47D细胞中表达降低(t=4. 341,P=0. 012),而miR-205在MDA-MB-231细胞中表达升高(t=-3. 266,P=0. 031)。结论 miR-130b的升高和miR-205的降低与乳腺癌的发生发展相关;miR-130b的升高和miR-205的降低可能是犀黄丸发挥抗癌作用途径之一。
Objective To study the differentially expressed microRNAs (miRNAs) in breast cancer formalin-fixed, paraffin-embedded (FFPE) tissue and the effect of Xihuang Pill extract on the expression of candidate miRNAs in breast cancer cell line in vitro. Methods Microarray was used to detect the differentially expressed miRNAs in breast cancer tissues, adjacent tissues and cancer tissues with different molecular types, and the results of unsupervised cluster analysis were processed by cluster software. Real-time fluorescence quantitative polymerase chain reaction (qPCR) was used to detect the expression of candidate miRNAs in 106 breast cancer tissues, 22 paracancerous tissues and 66 benigh breast lesions. The extract of Xihuang Pill interfered with T-47D cells and MDA-MB-231 cells, then the expression of candidate miRNAs in those cells were detected by qPCR. Results The microarray results showed that the expression of miR-130b was higher in cancer tissues than in adjacent tissues (P<0.05). The expression of miR-205 in basal-like cancer tissues with higher malignancy was significantly lower than that in luminal cancer tissues (P<0.05). qPCR showed that the expression miR-130b in cancer tissues (t=6.123, P=0.000) and adjacent tissues (t=-3.375, P=0.001) were significantly higher than that in benigh lesion tissues, and the expression of miR-205 in cancer tissues was significantly lower than that in benigh lesion tissues (t=-2.073, P=0.041). Cell experiment in vitro showed that the expression of miR-130b in T-47D cells decreased after the intervention of Xihuang Pill extract (t=4.341, P=0.012), while the expression of miR-205 in MDA-MB-231 cells increased after the intervention of Xihuang Pill extract (t=-3.266, P=0.031). Conclusions The increase of miR-130b and the decrease of miR-205 are related to the occurrence and development of breast cancer. The increase of miR-130b and the decrease of miR-205 may be the one of anti-tumor ways of Xihuang Pill.
Journal of Chinese Physician