Progesterone has been reported to inhibit the proliferation of breast cancer and osteosarcoma cells; however, its inhibitory mechanism has not yet been clarified. The aim of the present study was to clarify the effects of progesterone on apoptosis in breast cancer (MCF-7) and human osteosarcoma (MG-63) cells.
In this experimental study the cytotoxic effect of progesterone was measured in MCF-7 and MG-63 cells exposed to different concentrations of progesterone using MTT assay, and effective concentrations were identified. The expression levels of the Bax, P53 and Bcl-2 genes were evaluated by real-time PCR, and caspase-3, 8 and 9 activity levels were determined using a colorimetric method. Hoechst staining and flow cytometry were used to confirm apoptosis. The data were statistically analyzed using one-way analysis of variance (ANOVA) and independent-samples t-test.
Compared to the control group, we observed a significant increase in the expression levels of the Bax and P53 genes and the activity levels of caspase-3 and 9, and a significant decrease in the expression level of the Bcl-2 gene in MCF-7 and MG-63 treated with effective concentration of progesterone. The caspase-8 activity level did not change significantly in treated MG-63 but increased in treated MCF-7 cells. Hoechst staining and flow cytometry results confirmed apoptosis in the cells exposed to effective concentration of progesterone.
The cytotoxic effect of progesterone on breast cancer and osteosarcoma cells was mediated by apoptotic pathways. In this context, progesterone triggers the extrinsic and intrinsic apoptotic pathways in MCF-7 cells and induces the intrinsic apoptotic pathway in MG-63 cells.
Nagini S. Breast cancer: Current molecular therapeutic targets and new players. Anticancer Agents Med Chem 2017; 17: 152–63.
Kindblom L. Bone tumors: epidemiology, classification, pathology. In: Davies A, Sundaram M, James S, editors. Imaging of bone tumors and tumor-like lesions. Medical radiology. Berlin, Heidelberg: Springer; 2009. p. 1–15.
Gadducci A, Biglia N, Sismondi P, Genazzani AR. Breast cancer and sex steroids: critical review of epidemiological, experimental and clinical investigations on etiopathogenesis, chemoprevention and endocrine treatment of breast cancer. Gynecol Endocrinol 2005; 20: 343–60.
Farach-Carson MC, Lin S-H, Nalty T, Satcher RL. Sex differences and bone metastases of breast, lung, and prostate cancers: do bone homing cancers favor feminized bone marrow?. Front Oncol 2017; 7: 163.
Gonzalez Ricarte M, Castro Pérez Ad, Tarín JJ, Cano A. Progestogens and risk of breast cancer: a link between bone and breast?. Gynecol Endocrinol 2016; 32: 6–8.
Chaiwongwatanakul S, Yanatatsaneejit P, Tongsima S, Mutirangura A, Boonyaratanakornkit V. Sex steroids regulate expression of genes containing long interspersed elements-1s in breast cancer cells. Asian Pac J Cancer Prev 2016; 17: 4003–7.
Ruan X, Neubauer H, Yang Y, Schneck H, Schultz S, Fehm T, et al.. Progestogens and membrane-initiated effects on the proliferation of human breast cancer cells. Climacteric 2012; 15: 467–72.
Vares G, Ory K, Lectard B, Levalois C, Altmeyer-Morel S, Chevillard S, et al.. Progesterone prevents radiation-induced apoptosis in breast cancer cells. Oncogene 2004; 23: 4603.
Tian JM, Ran B, Zhang CL, Yan DM, Li XH. Estrogen and progesterone promote breast cancer cell proliferation by inducing cyclin G1 expression. Braz J Med Biol Res 2018; 51: 1–7.
Chen F-P, Chien M-H, Chen H-Y, Huang T-S, Ng Y-T. Effects of estradiol and progestogens on human breast cells: Regulation of sex steroid receptors. Taiwan J Obstet Gynecol 2013; 52: 365–73.
Im JY, Kim TH, Lee YJ, Kim IY, Kwack SJ, Byung Mu Lee CSM, et al.. Molecular mechanism of progesterone-induced apoptosis in human breast cancer T47D cells. Cancer prevention research 2008; 13: 177–83.
Luo G, Abrahams VM, Tadesse S, Funai EF, Hodgson EJ, Gao J, et al.. Progesterone inhibits basal and tnf-α-induced apoptosis in fetal membranes: a novel mechanism to explain progesterone-mediated prevention of preterm birth. Reprod Sci 2010; 17: 532–9.
Nguyen H, Syed V. Progesterone inhibits growth and induces apoptosis in cancer cells through modulation of reactive oxygen species. Gynecol Endocrinol 2011; 27: 830–6.
Kon A, Yuan B, Hanazawa T, Kikuchi H, Sato M, Furutani R, et al.. Contribution of membrane progesterone receptor α to the induction of progesterone-mediated apoptosis associated with mitochondrial membrane disruption and caspase cascade activation in Jurkat cell lines. Oncol Rep 2013; 30: 1965–70.
Wang Q-P, Xie H, Yuan L-Q, Luo X-H, Li H, Wang D, et al.. Effect of progesterone on apoptosis of murine MC3T3-E1 osteoblastic cells. Amino Acids 2009; 36: 57–63.
Yao W, Dai W, Shahnazari M, Pham A, Chen Z, Chen H, et al.. Inhibition of the progesterone nuclear receptor during the bone linear growth phase increases peak bone mass in female mice. PLoS One 2010; 5: e11410.
Chlebowski RT. Bone health in women with early-stage breast cancer. Clin Breast Cancer 2005; 5: S35–S40.
Bravo D, Shogren KL, Zuo D, Wagner ER, Sarkar G, Yaszemski MJ, et al.. 2‐methoxyestradiol‐mediated induction of frzb contributes to cell death and autophagy in MG-63 osteosarcoma cells. J Cell Biochem 2017; 118: 1497–504.
Rickard DJ, Iwaniec UT, Evans G, Hefferan TE, Hunter JC, Waters KM, et al.. Bone growth and turnover in progesterone receptor knockout mice. Endocrinology 2008; 149: 2383–90.
Liang M, Liao Ey, Xu X, Luo Xh, Xiao Xh. Effects of progesterone and 18‐methyl levonorgestrel on osteoblastic cells. Endocr Res 2003; 29: 483–501.
Dohi O, Hatori M, Suzuki T, Ono K, Hosaka M, Akahira JI, et al.. Sex steroid receptors expression and hormone‐induced cell proliferation in human osteosarcoma. Cancer Sci 2008; 99: 518–23.
Zhang N, Wang W, Li W, Liu C, Wang Y, Sun K. Reduction of progesterone, estradiol and hCG secretion by perfluorooctane sulfonate via induction of apoptosis in human placental syncytiotrophoblasts. Placenta 2015; 36: 575–80.
Li Y, Wang H, Zhou D, Shuang T, Zhao H, Chen B. Up-regulation of long noncoding rna sra promotes cell growth, inhibits cell apoptosis, and induces secretion of estradiol and progesterone in ovarian granular cells of mice. Med Sci Monit 2018; 24: 2384.
Chang W-T, Cheng H-L, Hsieh B-S, Chiu C-C, Lee K-T, Chang K-L. Progesterone increases apoptosis and inversely decreases autophagy in human hepatoma HA22T/VGH cells treated with epirubicin. Sci World J 2014: 2014.
Araki T, Yamamoto A, Yamada M. Accurate determination of DNA content in single cell nuclei stained with Hoechst 33258 fluorochrome at high salt concentration. Histochemistry 1987; 87: 331–8.
Yao G, Ling L, Luan J, Ye D, Zhu P. Nonylphenol induces apoptosis of Jurkat cells by a caspase-8 dependent mechanism. Int Immunopharmacol 2007; 7: 444–53.
Grott M, Karakaya S, Mayer F, Baertling F, Beyer C, Kipp M, et al.. Progesterone and estrogen prevent cisplatin-induced apoptosis of lung cancer cells. Anticancer Res 2013; 33: 791–800.
Hu Z, Deng X. The effect of progesterone on proliferation and apoptosis in ovarian cancer cell. Zhonghua Fu Chan Ke Za Zhi 2000; 35: 423–6.
Chávez-Riveros A, Garrido M, Apan MTR, Zambrano A, Díaz M, Bratoeff E. Synthesis and cytotoxic effect on cancer cell lines and macrophages of novel progesterone derivatives having an ester or a carbamate function at C-3 and C-17. Eur J Med Chem 2014; 82: 498–505.
Zhou L, Zhou W, Zhang H, Hu Y, Yu L, Zhang Y, et al.. Progesterone suppresses triple-negative breast cancer growth and metastasis to the brain via membrane progesterone receptor α. Int J Mol Med 2017; 40: 755–61.
Godbole M, Tiwary K, Badwe R, Gupta S, Dutt A. Progesterone suppresses the invasion and migration of breast cancer cells irrespective of their progesterone receptor status-a short report. Cell Oncol (Dordr) 2017; 40: 411–7.
MacNamara P, O'Shaughnessy C, Manduca P, Loughrey H. Progesterone receptors are expressed in human osteoblast-like cell lines and in primary human osteoblast cultures. Calcif Tissue Int 1995; 57: 436–41.
Fang D, Yang H, Lin J, Teng Y, Jiang Y, Chen J, et al.. 17beta-estradiol regulates cell proliferation, colony formation, migration, invasion and promotes apoptosis by upregulating miR-9 and thus degrades MALAT-1 in osteosarcoma cell MG-63 in an estrogen receptor-independent manner. Biochem Biophys Res Commun 2015; 457: 500–6.
Quinkler M, Kaur K, Hewison M, Stewart P, Cooper M. Progesterone is extensively metabolized in osteoblasts: implications for progesterone action on bone. Hormone Metab Res 2008; 40: 679–84.
Domínguez-Malagón HR, González-Conde E, Cano-Valdez AM, Luna-Ortiz K, Mosqueda-Taylor A. Expression of hormonal receptors in osteosarcomas of the jaw bones: Clinico-pathological analysis of 21 case. Medicina oral, patologia oral y cirugia bucal 2014; 19: e44.
Syed V, Ho S-M. Progesterone-induced apoptosis in immortalized normal and malignant human ovarian surface epithelial cells involves enhanced expression of FasL. Oncogene 2003; 22: 6883.
Han S, Lee K-M, Choi J-Y, Park SK, Lee J-Y, Lee JE, et al.. CASP8 polymorphisms, estrogen and progesterone receptor status, and breast cancer risk. Breast Cancer Res Treat 2008; 110: 387–93.
Pu X, Storr SJ, Zhang Y, Rakha EA, Green AR, Ellis IO, et al.. Caspase-3 and caspase-8 expression in breast cancer: caspase-3 is associated with survival. Apoptosis 2017; 22: 357–68.
Himuro T, Horimoto Y, Arakawa A, Matsuoka J, Tokuda E, Tanabe M, et al.. Activated caspase 3 expression in remnant disease after neoadjuvant chemotherapy may predict outcomes of breast cancer patients. Ann Surg Oncol 2016; 23: 2235–41.
Xiong J, Zhao J, Peng L, Wang H, Liang W. BRCA1 inhibits progesterone-induced proliferation and migration of breast cancer cells. Nan Fang Yi Ke Da Xue Xue Bao 2012; 32: 1105–10.