Pre-operative progesterone benefits operable breast cancer patients by modulating surgical stress
- 146 Downloads
We have reported a survival benefit of single injection of hydroxyprogesterone prior to surgery for primary tumour in patients with node-positive operable breast cancer. Hydroxyprogesterone was meant to recapitulate the luteal phase of menstrual cycle in these women. We wanted to understand the molecular basis of action of hydroxyprogesterone on primary breast tumours in a peri-operative setting.
We performed whole transcriptome sequencing (RNA-Seq) of primary breast tumour samples collected from patients before and after hydroxyprogesterone exposure and controls. Paired breast cancer samples were obtained from patients who were given hydroxyprogesterone before surgery and a group of patients who were subjected to only surgery.
A test of significance between the two groups revealed 207 significantly altered genes, after correction for multiple hypothesis testing. We found significantly contrasting gene expression patterns in exposed versus unexposed groups; 142 genes were up-regulated post-surgery among exposed patients, and down-regulated post-surgery among unexposed patients. Significantly enriched pathways included genes that respond to progesterone, cellular stress, nonsense-mediated decay of proteins and negative regulation of inflammatory response. These results suggest that cellular stress is modulated by hydroxyprogesterone. Network analysis revealed that UBC, a mediator of stress response, to be a major node to which many of the significantly altered genes connect.
Our study suggests that pre-operative exposure to progesterone favourably modulates the effect of surgical stress, and this might underlie its beneficial effect when administered prior to surgery.
KeywordsOperable breast cancer Hydroxyprogesterone RNA-Seq Network analysis
Authors would like to acknowledge Ms. Sriparna Biswas, Dr. Subrata Patra and Mr. Sumanta Sarkar for their help in performing the RNA-Seq laboratory experiments. We would like to thank Prof. Bidyut Roy, Mr. Badal De and Ms. Anindita Ray for helping in qPCR experiments. We would also like to thank Drs Nita Nair, Shalaka Joshi, Vani Parmar, Rohini Hawaldar and Vaibhav Vanmali for sample acquisition.
PP, PPM, RAB and SG conceived, designed and executed the study. RAB, RC, NG, AD and SG provided clinical samples for the study. SC and AM generated RNA-Seq data. SC, PP and PPM performed RNA-Seq data analysis. SC, RC, NG, AM, AD, PP, SG and PPM interpreted the results. SC, RC, NG, AD, SG, RAB, PPM and PP participated in writing of manuscript.
Authors acknowledge funding support from Department of Atomic Energy and Tata Memorial Centre for conducting the study. SC would like to acknowledge Indian Council for Medical Research for her doctoral research fellowship. RC is supported by a research fellowship from the Homi Bhabha National Institute (HBNI), Tata Memorial Centre. NG is supported by Prime Minister’s Fellowship Scheme for Doctoral Research, a public–private partnership between Science and Engineering Research Board (SERB), Department of Science and Technology, Government of India and Confederation of Indian Industry (CII). PPM was supported by the J. C. Bose Fellowship of the Government of India, Department of Science and Technology.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This study was approved by the Institutional Ethics Committees of the Tata Memorial Centre, Mumbai, and the National Institute of Biomedical Genomics, Kalyani. Biospecimens were collected from breast cancer patients with written informed consent.
- 3.Lemon HM, Rodriguez-Sierra JF (1996) Timing of breast cancer surgery during the luteal menstrual phase may improve prognosis. Nebraska Med J 81(3):73–78Google Scholar
- 5.Badwe R, Hawaldar R, Parmar V, Nadkarni M, Shet T, Desai S, Gupta S, Jalali R, Vanmali V, Dikshit R, Mittra I (2011) Single-injection depot progesterone before surgery and survival in women with operable breast cancer: a randomized controlled trial. J Clin Oncol 29(21):2845–2851. https://doi.org/10.1200/jco.2010.33.0738 CrossRefPubMedGoogle Scholar
- 6.Trapnell C, Roberts A, Goff L, Pertea G, Kim D, Kelley DR, Pimentel H, Salzberg SL, Rinn JL, Pachter L (2012) Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks. Nat Protoc 7(3):562–578. https://doi.org/10.1038/nprot.2012.016 CrossRefPubMedPubMedCentralGoogle Scholar
- 8.Trapnell C, Williams BA, Pertea G, Mortazavi A, Kwan G, van Baren MJ, Salzberg SL, Wold BJ, Pachter L (2010) Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation. Nat Biotechnol 28(5):511–515. https://doi.org/10.1038/nbt.1621 CrossRefPubMedPubMedCentralGoogle Scholar
- 9.Benjamini Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J Roy Stat Soc 57(1):289–300Google Scholar
- 10.Kuleshov MV, Jones MR, Rouillard AD, Fernandez NF, Duan Q, Wang Z, Koplev S, Jenkins SL, Jagodnik KM, Lachmann A, McDermott MG, Monteiro CD, Gundersen GW, Ma’ayan A (2016) Enrichr: a comprehensive gene set enrichment analysis web server 2016 update. Nucleic Acids Res 44(W1):W90–W97. https://doi.org/10.1093/nar/gkw377 CrossRefPubMedPubMedCentralGoogle Scholar
- 14.Glenisson M, Vacher S, Callens C, Susini A, Cizeron-Clairac G, Le Scodan R, Meseure D, Lerebours F, Spyratos F, Lidereau R, Bieche I (2012) Identification of new candidate therapeutic target genes in triple-negative breast cancer. Genes Cancer 3(1):63–70. https://doi.org/10.1177/1947601912449832 CrossRefPubMedPubMedCentralGoogle Scholar
- 20.Sun X, Glynn DJ, Hodson LJ, Huo C, Britt K, Thompson EW, Woolford L, Evdokiou A, Pollard JW, Robertson SA, Ingman WV (2017) CCL2-driven inflammation increases mammary gland stromal density and cancer susceptibility in a transgenic mouse model. Breast Cancer Research 19(1):4. https://doi.org/10.1186/s13058-016-0796-z CrossRefPubMedPubMedCentralGoogle Scholar
- 35.Ryu KY, Maehr R, Gilchrist CA, Long MA, Bouley DM, Mueller B, Ploegh HL, Kopito RR (2007) The mouse polyubiquitin gene UbC is essential for fetal liver development, cell-cycle progression and stress tolerance. EMBO J 26(11):2693–2706. https://doi.org/10.1038/sj.emboj.7601722 CrossRefPubMedPubMedCentralGoogle Scholar