Effect of 3α-dihydroprogesterone and 5α-dihydroprogesterone on DCIS cells and possible impact for postmenopausal women

Abstract

Objective

Progesterone metabolites 5α-dihydroprogesterone (5αP) and 3α-dihydroprogesterone (3αP) have opposite effects on proliferation, apoptosis and metastasis in the breast. Evidence regarding their influence on ductal carcinoma in situ (DCIS) lesions is lacking.

Methods

MCF10DCIS.com cells were cultured in a 3D culture system and treated with 5αP or 3αP. After 5 and 12 days of treatment, polymerase chain reaction (PCR) of proliferation, invasion/metastasis, anti-apoptotic or other markers was performed. Cells treated with the tumor-promoting 5αP were observed under the light and confocal microscopes to reveal possible morphological changes that could indicate a transition from an in situ to an invasive phenotype. As a control, the morphology of the MDA-MB-231 invasive cell line was examined. The invasive potential after exposure to 5αP was also assessed using a detachment assay.

Results

The PCR analysis of the chosen markers showed no statistically significant difference between naive cells and cells treated with 5αP or 3αP. DCIS spheroids retained their in situ morphology after treatment with 5αP. The detachment assay showed no increased potential for invasion after exposure to 5αP. Progesterone metabolites 5αP and 3αP do not facilitate or prohibit tumor promotion/invasion in MCF10DCIS.com cells, respectively.

Conclusion

As oral micronized progesterone has been proved effective for hot flushes in postmenopausal women, first in vitro data propose that progesterone-only therapy could possibly be considered for women after DCIS suffering from hot flushes.

摘要

目的：孕酮代谢产物5α-二氢孕酮(5αP)和3α-二氢孕酮(3αP)对乳腺细胞增殖、凋亡和转移的影响相反。关于其对导管原位癌(DCIS)病变影响的证据尚缺乏。

方法：MCF10DCIS.com细胞在3D培养体系中培养, 分别用5αP或3αP处理。治疗5、12 d后, 采用聚合酶链式反应(PCR)检测细胞增殖、侵袭、转移、抗凋亡等标志物。用促进肿瘤的5αP处理的细胞在光和共聚焦显微镜下观察, 以揭示可能的形态变化, 这些变化可能表明从原位表型转变为侵袭性表型。作为对照, 检测MDA-MB-231侵袭细胞系的形态。暴露于5aP后的侵袭潜能也采用分离试验进行评估。

结果：PCR对所选标记进行分析结果显示, 未处理细胞与经5αP和3αP处理的细胞之间无统计学差异。5αP处理后, DCIS保持原位形态。分离试验显示暴露于5αP后没有增加侵袭性。孕酮代谢物5αP和3αP不促进或者抑制MCF10DCIS.com细胞的肿瘤进展、侵袭。

结论：由于口服微孕酮已被证明对绝经后女性潮热有效, 首次体外数据表明, 可考虑仅应用孕酮治疗DCIS后女性潮热。

Keywords:

• Ductal carcinoma in situ
• progesterone metabolites
• postmenopause

Acknowledgements

This work was partially conducted during participation in the mentoring program Campus GE by BESINS Health. With Campus GE, a mentor–mentee program based on essential professional skills was initiated, where Ludwig Kiesel as a mentor and Marina Sourouni as a mentee worked together. However, BESINS Health had no role in the design of the project, analyses or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Potential conflict of interest

The authors report no conflict of interests. The authors alone are responsible for the content and writing of the article.

Source of funding

This research received no external funding.