高通量测序揭秘中药苦参如何杀死乳腺癌细胞

　　2016年9月1日，美国《肿瘤标靶》在线发表澳大利亚阿德莱德大学、山西中医学院的研究报告，从中国传统草药中得到了复杂混合物，发现其具有杀死乳腺癌细胞的作用，解开了来自一种传统中药的植物化合物复杂混合如何起作用杀死癌细胞，首次描述了传统中药的分子作用，而不是将其分解成其组成部分。

　　在中国，复方苦参注射液被批准用于治疗各种癌症，通常用作辅助西药化疗，但是这种药物的抗癌原理尚未得知。这是第一次有研究指出，植物来源化合物复杂混合物的分子作用方式。

　　大多数中药都是基于其在中国数百年或上千年的使用经验。通常有大量的证据表明，中药在一些疾病的治疗过程中疗效确实不错，但是没有人知道这些药物治疗疾病的具体现代药理学机制。经试验，如果打破中药药方中的成分，分解和测试许多传统中药的成分，会发现单个化合物并没有太多的活性，不会起到很好的治疗效果，通常一些药物的组合才能够显效。多种化合物的组合可能有效，并也意味着很少的副作用。该研究的苦参注射液成分十分复杂，是运用所谓的系统生物学方法，从两种药材（苦参和白土苓）根部提取而来。这是一种分析复杂生物系统的方法，试图考虑系统的所有可测量方面，而不是专注于一个单一的变量。

　　该研究在实验室中利用乳腺癌细胞，采用高通量的新一代测序技术，对复方苦参注射液影响的生物信号通路进行了测定，识别其所靶定的基因和生物通路，发现复方苦参注射液影响了西医化疗中具有共性的若干基因。复方苦参注射液触发的基因表达模式，可影响与西方化疗相同的通路，但是作用于同一通路中的不同基因。这些受影响的基因调节细胞的分裂、生存周期和死亡，并且复方苦参注射液改变了细胞周期被调控的方式，以将癌细胞推向细胞死亡途径，从而杀死细胞，似乎是复方苦参注射液杀死癌细胞的可能机制。这项技术可以用来对其他中国传统药物起效的具体分子生物学机制进行现代化研究，为中药现代化提供新的思路，可能会打开其在西方医学中的使用方式。

Oncotarget. 2016 Sep 1. [Epub ahead of print]

Identification of candidate anti-cancer molecular mechanisms of compound kushen injection using functional genomics.

Qu Z, Cui J, Harata-Lee Y, Aung TN, Feng Q, Raison JM, Kortschak RD, Adelson DL.

The University of Adelaide, Adelaide, South Australia 5005, Australia; Shanxi University of Traditional Chinese Medicine, Shanxi 030619, China.

Compound Kushen Injection (CKI) has been clinically used in China for over 15 years to treat various types of solid tumours. However, because such Traditional Chinese Medicine (TCM) preparations are complex mixtures of plant secondary metabolites, it is essential to explore their underlying molecular mechanisms in a systematic fashion. We have used the MCF-7 human breast cancer cell line as an initial in vitro model to identify CKI induced changes in gene expression. Cells were treated with CKI for 24 and 48 hours at two concentrations (1 and 2 mg/mL total alkaloids), and the effect of CKI on cell proliferation and apoptosis were measured using XTT and Annexin V/Propidium Iodide staining assays respectively. Transcriptome data of cells treated with CKI or 5-Fluorouracil (5-FU) for 24 and 48 hours were subsequently acquired using high-throughput Illumina RNA-seq technology. In this report we show that CKI inhibited MCF-7 cell proliferation and induced apoptosis in a dose-dependent fashion. We integrated and applied a series of transcriptome analysis methods, including gene differential expression analysis, pathway over-representation analysis, de novo identification of long non-coding RNAs (lncRNA) as well as co-expression network reconstruction, to identify candidate anti-cancer molecular mechanisms of CKI. Multiple pathways were perturbed and the cell cycle was identified as the potential primary target pathway of CKI in MCF-7 cells. CKI may also induce apoptosis in MCF-7 cells via a p53 independent mechanism. In addition, we identified novel lncRNAs and showed that many of them might be expressed as a response to CKI treatment.

KEYWORDS: lncRNA; systems biology; traditional chinese medicine; transcriptome

PMID: 27602759

DOI: 10.18632/oncotarget.11788