Differential control of growth, cell cycle progression, and expression of NF-κB in human breast cancer cells MCF-7, MCF-10A, and MDA-MB-231 by ponicidin and oridonin, diterpenoids from the chinese herb Rabdosia rubescens

doi:10.1016/j.bbrc.2005.09.040

Biochemical and Biophysical Research Communications

Volume 337, Issue 1, 11 November 2005, Pages 224-231

https://doi.org/10.1016/j.bbrc.2005.09.040 Get rights and content

Abstract

Ponicidin and oridonin are novel diterpenoids isolated from Rabdosia rubescens. We tested their effects in MCF-7 and MDA-MB-231 cells, as representing low and high invasive breast carcinoma, with normal MCF-10A cells. Clonogenicity and proliferation in MCF-7 cells were inhibited more significantly by ponicidin than oridonin, while the reverse was observed in MCF-10A cells. Ponicidin and oridonin induced S/G₂M arrest and G₁/S block in MCF-7 cells. In MCF-10A cells treated with either diterpenoid, induction of apoptosis was observed. Moreover, oridonin almost completely blocked MCF-10A progression from S to G₂/M phase; in contrast, ponicidin-treated MCF-10A cells showed no discernable changes in cell cycle phase distribution. Neither diterpenoid affected growth of MDA-MB-231 cells, at the dose range effective for MCF-7 or MCF-10A cells. Ponicidin-treated MCF-7 cells expressed reduced levels of cyclin B1, cdc2, transcription factor E2F, and Rb including phosphorylation at S780. Less pronounced effects were found in cells treated with oridonin. Neither compound altered cyclin D1 and cdk4 in MCF-7 cells. In MCF-10A cells, oridonin was more active than ponicidin in inhibiting the expression of cyclin B1, cdc2, S780-phosphorylated Rb, and E2F. To further investigate induction of apoptosis in MCF-10A cells, we measured changes in NF-κB. Decreases in p65 or p50 forms of NF-κB and its upstream regulator I-κB were found in oridonin-treated MCF-10A and not MCF-7 cells. Taken together, these results provide a mechanistic framework for the cellular effects of ponicidin and oridonin in different stage breast cancer cells.

Section snippets

Materials and methods

Cell cultures. Breast adenocarcinoma cell lines were purchased from American Type Culture Conditions (ATCC, Rockville, MD) and cultured using the following conditions. MCF-7 cells were maintained in Eagle’s minimum essential medium supplemented with 2 mM glutamine and Earle’s BSS adjusted to contain 1.5 g/L sodium bicarbonate, 0.1 mM non-essential amino acids, and 1 mM sodium pyruvate, and supplemented with 0.01 mg/ml bovine insulin and 10% fetal bovine serum. MCF-10A cells were maintained in MEGM

Effects of ponicidin and oridonin on colony formation in MCF-7 cells

We first determined the effects of ponicidin and oridonin on colony formation, also referred to as clonogenicity, in MCF-7, MCF-10 A, and MDA-MB-231 cells. This assay measures the ability of tumor cells to grow and form foci, while normal cells become growth contact inhibited. Clonogenicity is therefore an indirect estimation of the propensity of tumor cells to undergo neoplastic transformation. Clonogenicity was determined by plating a fixed number of MCF-7 cells onto multiple well tissue

Discussion

Rabdosia rubescens is one of several Chinese herbs present in the once popular dietary supplement, denoted PC-SPES. Use of the entire plant of this Chinese herb purportedly shows digestive benefit systems and anecdotal evidence of efficacy for esophageal carcinoma, with little toxicity [27]. The main active components of R. rubescens are tetracyclic diterpenoid compounds, such as oridonin and ponicidin [28]. More recently, one of its active constituents, oridonin has been evaluated as adjunct

Acknowledgment

This research was supported in part by NCI Clinical Nutrition Research Unit Grant CA 29502 to T.C.H.

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Differential control of growth, cell cycle progression, and expression of NF-κB in human breast cancer cells MCF-7, MCF-10A, and MDA-MB-231 by ponicidin and oridonin, diterpenoids from the chinese herb Rabdosia rubescens

Abstract

Section snippets

Materials and methods

Effects of ponicidin and oridonin on colony formation in MCF-7 cells

Discussion

Acknowledgment

J. Nutr.

Eur. J. Cancer

Biochem. Biophys. Res. Commun.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

Cell

J. Pharmacol. Sci.

Surg. Oncol.

Epidemiology. Search for a killer: focus shifts from fat to hormones

Science

Multi-ethnic differences in breast cancer: current concepts and future directions

Int. J. Cancer

Cancer statistics, 2002, CA

Cancer J. Clin.

Estimating the world cancer burden: Globocan 2000

Int. J. Cancer

Mammary gland development, reproductive history, and breast cancer risk

Cancer Res.

Steroid hormone receptor signaling in tumorigenesis

J. Cell Biochem.

Energetic factors, ovarian steroids and the risk of breast cancer

Eur. J. Cancer Prev.

A systematic overview of chemotherapy effects in breast cancer

Acta Oncol.

Alternative therapies used by women with breast cancer in four ethnic populations

J. Natl. Cancer Inst.

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Nat. Rev. Cancer

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Carcinogenesis

Chinese herbal mixture PC SPES in treatment of prostate cancer (review)

Int. J. Oncol.