Estrone and progesterone inhibit the growth of murine MC38 colon cancer line

doi:10.1016/j.jsbmb.2008.11.007

The Journal of Steroid Biochemistry and Molecular Biology

Volume 113, Issues 1–2, January 2009, Pages 75-79

https://doi.org/10.1016/j.jsbmb.2008.11.007 Get rights and content

Abstract

The unsatisfactory effectiveness of reference chemotherapy in colon cancer (fluorouracil – FU) results in continuous search for agents, which could enhance the action of FU. Some epidemiological data such as a decreased risk of colorectal cancer among menopausal women receiving hormonal replacement therapy indicate the role of female sex hormones in the pathogenesis of this disease.

The aim of this study was to examine the direct effects of various concentrations of estrone and progesterone (10⁻⁴ to 10⁻¹² M) applied alone or together with FU on the growth of murine MC38 colon cancer in vitro.

Estrone inhibited MC38 cancer growth in a wide range of concentrations (10⁻¹² to 10⁻⁴ M) with similar potency and at some concentrations (10⁻⁶ and 10⁻⁴ M) augmented also the cytotoxic action of FU. Progesterone induced MC38 cancer growth inhibition at high concentrations (10⁻⁵ to 10⁻⁴ M) in dose- and time-dependent manner but it did not intensify antineoplastic effect of FU. A weak inhibitory effect of progesterone was also observed for lower concentrations (10⁻⁵ to 10⁻¹⁰ M) in long lasting cultures (72 h).

The results indicate that estrone and progesterone inhibit the MC38 cancer growth and that estrone increases also the cytotoxic effect of FU, what confirms the role of female sex steroids in modulation of colon cancer growth.

Introduction

In the developed countries colorectal cancer is one of the most common cancers [1]. Chemotherapy is an important part of treatment in this neoplasm, because approximately 30% of all patients with colon cancer have metastatic disease at diagnosis, and 50% of early-stage patients will eventually develop metastatic or advanced disease [2]. For about 50 years, fluorouracil (FU) has remained the main chemotherapeutic agent in this cancer. Unfortunately, the response to FU is observed only in 10–15% patients [3]. Its combination with leucovorin (LV) enhanced the mean response ratio to 23% and became the standard regimen in adjuvant and palliative chemotherapy of this disease [4]. During the last decade FDA (Food and Drug Administration) has approved six new drugs for treatment of advanced stages of colon cancer. Three of them belong to cytotoxic agents: irinotecan (1996), oxaliplatin (2002), oral formulation of fluorouracil – capecitabine (1998), and the other three to monoclonal antibodies: bevacizumab targeting vascular endothelial growth factor (2004), cetuximab (2004) [5] and panitumumab (2006) [6], both directed against the epithelial growth factor receptor. Although the new drugs prolonged the median survival from 12 months (FU + LV) to about 21 months, the effectiveness of the therapy is still unsatisfactory and the treatment of metastatic disease remains palliative [5]. Therefore, there are still researches carried on for new substances enhacing the antineoplastic effect of FU, including cytostatic drugs as well as other biomodulators.

The role of estrogens in colon carcinogenesis has been discussed for many years. This association was suggested by some epidemiological data such as an age-specific occurrance of colorectal cancer in women [7], protective influence of increasing parity and sex differences in site specific incidences of the neoplastic lesion in bowels [8]. Moreover, a lot of observational studies [9] and the last randomized primary prevention trial – the Women's Health Initiative (WHI) [10] showed a decreased risk of colorectal cancer among menopausal women receiving hormonal replacement therapy (HRT). Although generally most of these protective effects are attributed to the action of estrogens, it could not be excluded that the positive effects are connected also with the other female sex hormone – progesterone. Such a hypothesis could be supported by the fact that in the WHI study the reduction in colon cancer risk was observed only in users of combined estrogen and progestin HRT [10], whereas in women after hysterectomy, receiving only estrogens, the protective effect was not noticed [11].

In spite of many facts implying involvement of female sex steroids in colorectal carcinogenesis, the number of studies examining the direct effect of these hormones on the colon cancer growth is limited and gives often opposing results. Studies in vitro [12] and experiments with ovariectomized animals [13], [14] have shown that estradiol (E2), which is the most often examined estrogen, can inhibit as well as stimulate the growth of this neoplasm. The influence of the weaker estrogen – estrone (E1) and progesterone on the colon cancer growth has been hardly ever a matter of study.

Thus, the aim of this paper was to examine the direct effects of estrone and progesterone applied alone or together with FU on the murine MC38 colon cancer growth in vitro assessed by two colorimetric methods reflecting changes in proliferation and apoptosis.

Section snippets

Materials and methods

Murine Colon 38 cancer cells were used in the study. The cells were routinely grown in a humidified incubator at 37 °C with 5% CO₂ in RPMI 1640 medium (Sigma), supplemented with: 25 nM Hepes buffer (Sigma), 4 mM L-glutamine (Sigma), 100 U/ml penicillin and 100 μg/ml streptomycin solution (Sigma), 2 g/l sodium bicarbonate (Sigma) and 5% fetal calf serum (FCS, Biochrom). The cells were passaged every 7 days with 0.05% trypsin/0.02% EDTA (Trypsin-EDTA, Sigma) and the medium was changed every 3–4 days.

Results

Estrone in a wide range of concentrations (10⁻¹² to 10⁻⁴ M) induced similar, moderately strong inhibition of the MC38 cancer growth, which was slightly more potent after 72 h (up to 72% of control group) than after 24 h incubation (up to 81% of control group) (Fig. 1). The inhibitory effect of estrone was not observed only for the concentration of 10⁻⁶ M in 24 h culture and for the concentration of 10⁻⁵ M in 72 h culture (Fig. 1). The beginning of action was noticed as early as after 4 h incubation in

Discussion

In the present study we have shown that estrone and progesterone inhibited MC38 colon cancer growth. To our knowledge, we have also revealed for the first time that estrone can potentiate the cytotoxic action of FU. In our study, estrone induced MC38 growth inhibition in a wide range of concentrations (10⁻¹²–10⁻⁴ M) with the exception of 10⁻⁶ M in 24 h culture and 10⁻⁵ M in 72 h culture. Such an effect could be connected with a different mechanism of action for the lower (i.e. under the ineffective

Conclusions

Summing up, we have shown in this paper that estrone and progesterone inhibited MC38 colon cancer growth and that estrone intensified cytotoxic effect of FU. Our results indicate that female sex steroids are involved in modulation of colon cancer growth. It suggests the potential use of these hormones, especially estrone, in colon cancer therapy, which could enhance the efficacy of FU. Further studies are needed to elucidate whether this hypothesis is true in human colon cancer and whether it

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Estrone and progesterone inhibit the growth of murine MC38 colon cancer line

Abstract

Introduction

Section snippets

Materials and methods

Results

Discussion

Conclusions

Am. J. Med.

Biochem. Biophys. Res. Commun.

Gastroenterology

J. Steroid. Biochem. Mol. Biol.

J. Nutr.

Cancer Lett.

Global cancer statistics 2002

C. A. Cancer J. Clin.

Metastatic colorectal cancer: systemic treatment in the new millennium

Cancer Control.

5-Fluorouracil: mechanisms of action and clinical strategies

Nat. Rev. Cancer

Colorectal cancer