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Gene expression profiling reveals effects of Cimicifuga racemosa (L.) NUTT. (black cohosh) on the estrogen receptor positive human breast cancer cell line MCF-7.

Gaube F, Wolfl S, Pusch L, Kroll TC, Hamburger M - BMC Pharmacol. (2007)

Bottom Line: Also several transcripts coding for oxidoreductases were induced, as for example the cytochrome P450 family members 1A1 and 1B1.In addition, some transcripts associated with antitumor but also tumor-promoting activity were regulated.The effects may be results of the activation of different pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Pharmacy, Department of Pharmaceutical Biology, University of Jena, Semmelweisstr, 10, 07743 Jena, Germany. Friedemann.Gaube@uni-jena.de

ABSTRACT

Background: Extracts from the rhizome of Cimicifuga racemosa (black cohosh) are increasingly popular as herbal alternative to hormone replacement therapy (HRT) for the alleviation of postmenopausal disorders. However, the molecular mode of action and the active principles are presently not clear. Previously published data have been largely contradictory. We, therefore, investigated the effects of a lipophilic black cohosh rhizome extract and cycloartane-type triterpenoids on the estrogen receptor positive human breast cancer cell line MCF-7.

Results: Both extract and purified compounds clearly inhibited cellular proliferation. Gene expression profiling with the extract allowed us to identify 431 regulated genes with high significance. The extract induced expression pattern differed from those of 17beta-estradiol or the estrogen receptor antagonist tamoxifen. We observed a significant enrichment of genes in an anti-proliferative and apoptosis-sensitizing manner, as well as an increase of mRNAs coding for gene products involved in several stress response pathways. These functional groups were highly overrepresented among all regulated genes. Also several transcripts coding for oxidoreductases were induced, as for example the cytochrome P450 family members 1A1 and 1B1. In addition, some transcripts associated with antitumor but also tumor-promoting activity were regulated. Real-Time RT-PCR analysis of 13 selected genes was conducted after treatment with purified compounds - the cycloartane-type triterpene glycoside actein and triterpene aglycons - showing similar expression levels compared to the extract.

Conclusion: No estrogenic but antiproliferative and proapoptotic gene expression was shown for black cohosh in MCF-7 cells at the transcriptional level. The effects may be results of the activation of different pathways. The cycloartane glycosides and - for the first time - their aglycons could be identified as an active principle in black cohosh.

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Cell cycle pathway diagram obtained from GenMAPP (Gene Map Annotator and Pathway Profiler, Gladstone Institutes, University of San Francisco, San Francisco, CA [38]. Proteins involved in cell cycle control are displayed from left to right as cell cycle progresses from G1 through S and G2 to M-phase. Genes with boxes marked black were upregulated. Genes marked with grey boxes were downregulated.
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Figure 5: Cell cycle pathway diagram obtained from GenMAPP (Gene Map Annotator and Pathway Profiler, Gladstone Institutes, University of San Francisco, San Francisco, CA [38]. Proteins involved in cell cycle control are displayed from left to right as cell cycle progresses from G1 through S and G2 to M-phase. Genes with boxes marked black were upregulated. Genes marked with grey boxes were downregulated.

Mentions: In agreement with the anti-proliferative effect of the black cohosh extract, genes involved in proliferation control were significantly overrepresented. Transcripts related to cell cycle regulation and DNA replication were regulated in a manner supporting cell cycle arrest. Genes, whose products are involved in the transition from G1 to S-phase appeared to be downregulated, such as cyclins (CCNA2, CCNE2, CCNF), cyclin-dependent kinase 2 (CDK2) and transcription regulators (E2F2, PCNA, SKP2), whereas transcription of inhibitory genes cyclin G2 (CCNG2), GADD45A (growth arrest and DNA-damage-inducible, alpha) and p21cip1 (cyclin-dependent kinase inhibitor 1A, CDKN1A) was increased. Elevated levels of CCNG2, cyclin B1 interacting protein 1 (CCNB1IP1), forkhead box O3A (FOXO3A), GADD45A and p21cip1 genes as well as downregulation of cyclin A2 (CCNA2) and CDK2 provided evidence that cell cycle progression might be additionally arrested at the G2/M-checkpoint. The level of various DNA replication related genes (CDC6, CDT1, FEN1, MCM2, MCM3, MCM4, MCM5, MCM7, MCM10, Pfs2, RFC3) was also reduced, thereby suggesting a reduction in the replication rate. All cell cycle related effects are summarized in Figure 5, which shows a cell cycle diagram from GenMAPP (Gene Map Annotator and Pathway Profiler, Gladstone Institutes, University of San Francisco, San Francisco, CA) [38], in which up- and downregulated genes are marked.


Gene expression profiling reveals effects of Cimicifuga racemosa (L.) NUTT. (black cohosh) on the estrogen receptor positive human breast cancer cell line MCF-7.

Gaube F, Wolfl S, Pusch L, Kroll TC, Hamburger M - BMC Pharmacol. (2007)

Cell cycle pathway diagram obtained from GenMAPP (Gene Map Annotator and Pathway Profiler, Gladstone Institutes, University of San Francisco, San Francisco, CA [38]. Proteins involved in cell cycle control are displayed from left to right as cell cycle progresses from G1 through S and G2 to M-phase. Genes with boxes marked black were upregulated. Genes marked with grey boxes were downregulated.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2194763&req=5

Figure 5: Cell cycle pathway diagram obtained from GenMAPP (Gene Map Annotator and Pathway Profiler, Gladstone Institutes, University of San Francisco, San Francisco, CA [38]. Proteins involved in cell cycle control are displayed from left to right as cell cycle progresses from G1 through S and G2 to M-phase. Genes with boxes marked black were upregulated. Genes marked with grey boxes were downregulated.
Mentions: In agreement with the anti-proliferative effect of the black cohosh extract, genes involved in proliferation control were significantly overrepresented. Transcripts related to cell cycle regulation and DNA replication were regulated in a manner supporting cell cycle arrest. Genes, whose products are involved in the transition from G1 to S-phase appeared to be downregulated, such as cyclins (CCNA2, CCNE2, CCNF), cyclin-dependent kinase 2 (CDK2) and transcription regulators (E2F2, PCNA, SKP2), whereas transcription of inhibitory genes cyclin G2 (CCNG2), GADD45A (growth arrest and DNA-damage-inducible, alpha) and p21cip1 (cyclin-dependent kinase inhibitor 1A, CDKN1A) was increased. Elevated levels of CCNG2, cyclin B1 interacting protein 1 (CCNB1IP1), forkhead box O3A (FOXO3A), GADD45A and p21cip1 genes as well as downregulation of cyclin A2 (CCNA2) and CDK2 provided evidence that cell cycle progression might be additionally arrested at the G2/M-checkpoint. The level of various DNA replication related genes (CDC6, CDT1, FEN1, MCM2, MCM3, MCM4, MCM5, MCM7, MCM10, Pfs2, RFC3) was also reduced, thereby suggesting a reduction in the replication rate. All cell cycle related effects are summarized in Figure 5, which shows a cell cycle diagram from GenMAPP (Gene Map Annotator and Pathway Profiler, Gladstone Institutes, University of San Francisco, San Francisco, CA) [38], in which up- and downregulated genes are marked.

Bottom Line: Also several transcripts coding for oxidoreductases were induced, as for example the cytochrome P450 family members 1A1 and 1B1.In addition, some transcripts associated with antitumor but also tumor-promoting activity were regulated.The effects may be results of the activation of different pathways.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Pharmacy, Department of Pharmaceutical Biology, University of Jena, Semmelweisstr, 10, 07743 Jena, Germany. Friedemann.Gaube@uni-jena.de

ABSTRACT

Background: Extracts from the rhizome of Cimicifuga racemosa (black cohosh) are increasingly popular as herbal alternative to hormone replacement therapy (HRT) for the alleviation of postmenopausal disorders. However, the molecular mode of action and the active principles are presently not clear. Previously published data have been largely contradictory. We, therefore, investigated the effects of a lipophilic black cohosh rhizome extract and cycloartane-type triterpenoids on the estrogen receptor positive human breast cancer cell line MCF-7.

Results: Both extract and purified compounds clearly inhibited cellular proliferation. Gene expression profiling with the extract allowed us to identify 431 regulated genes with high significance. The extract induced expression pattern differed from those of 17beta-estradiol or the estrogen receptor antagonist tamoxifen. We observed a significant enrichment of genes in an anti-proliferative and apoptosis-sensitizing manner, as well as an increase of mRNAs coding for gene products involved in several stress response pathways. These functional groups were highly overrepresented among all regulated genes. Also several transcripts coding for oxidoreductases were induced, as for example the cytochrome P450 family members 1A1 and 1B1. In addition, some transcripts associated with antitumor but also tumor-promoting activity were regulated. Real-Time RT-PCR analysis of 13 selected genes was conducted after treatment with purified compounds - the cycloartane-type triterpene glycoside actein and triterpene aglycons - showing similar expression levels compared to the extract.

Conclusion: No estrogenic but antiproliferative and proapoptotic gene expression was shown for black cohosh in MCF-7 cells at the transcriptional level. The effects may be results of the activation of different pathways. The cycloartane glycosides and - for the first time - their aglycons could be identified as an active principle in black cohosh.

Show MeSH
Related in: MedlinePlus