In HEK293 cells expressing human ERα, levonorgestrel was a full reporter agonist with lower potency than estradiol. This overexpression result should not be treated as evidence of clinically important estrogenic activity. [2]
In mouse mammary organoids, levonorgestrel increased Rankl and Wnt4 transcripts. Enzalutamide reduced the Rankl response at 100 µM, while Wnt4 induction was not significantly affected; the Rankl trend at 10 µM was not significant. [3]
PR-B-dependent reporter activation
Cell-based study · COS-1 monkey kidney cells transiently expressing human PR-B and pTAT-2xPRE-E1b-luciferase; empty-vector control [1]
LNG was a full PR-B agonist relative to R5020 in this assay. Reporter activation was negligible without transfected PR-B. Efficacy is distinct from potency; results do not supply a universal binding IC50.
Literature-Reported Treatment Conditions
- Treatment concentration
- Increasing ligand concentrations (dose-response; Figure 1)
- Exposure duration
- 24 hours
- Method
- Luciferase normalized to protein; transfection condition #1
- Vehicle
- 0.1% EtOH
- Reported limits
- No exact EC50 transcribed: supplementary numeric tables were not retrieved. The displayed curves are not used to estimate decimals.
Methods and source locations
Compound reported in the paper: Levonorgestrel; Compound label in the paper: LNG
Reported levonorgestrel material and experimental system; no verification of Adooq A10529 or lot equivalence.
Sections 2.1–2.4 and 3.1; Figure 1A–D and caption
PR-B reporter transactivation: R5020 maximum set to 100%; at least three independent experiments, each in triplicate; error bars SEM — Sections 2.1–2.4 and 3.1; Figure 1A–D and caption
Androgen-receptor reporter agonism
Cell-based study · COS-1 cells expressing human AR and pTAT-2xPRE-E1b-luciferase [2]
LNG activated the AR reporter with agonist efficacy and potency reported as similar to DHT under these conditions. Receptor-free transcriptional activity was negligible (reported data not shown).
Literature-Reported Treatment Conditions
- Treatment concentration
- Increasing concentrations (agonist dose-response)
- Exposure duration
- 24 hours
- Method
- Luciferase normalized to protein; MIB reference agonist
- Reported limits
- Vehicle identity/concentration for Figure 2 is not specified in its caption or assay subsection. EtOH explicitly specified for Figure 3 is not transferred here. Supplementary EC50 table not retrieved.
Methods and source locations
Compound reported in the paper: Levonorgestrel; Compound label in the paper: LNG
Reported levonorgestrel material and experimental system; no verification of Adooq A10529 or lot equivalence.
Sections 2.1–2.6 and 3.2; Figure 2A–C and caption
AR-mediated reporter transactivation: MIB maximum set as 100%; pooled independent experiments performed in triplicate, general methods specify at least two; SEM — Sections 2.1–2.6 and 3.2; Figure 2A–C and caption
Estrogen-receptor-alpha reporter agonism
Cell-based study · HEK293 human embryonic kidney cells expressing human ERα and pGL3-2xERE-pS2-luciferase [2]
LNG was a full agonist for ERα reporter transactivation but less potent than estradiol. This is a receptor-overexpression assay and is not evidence of clinically important estrogenic activity.
Literature-Reported Treatment Conditions
- Treatment concentration
- Increasing concentrations (agonist dose-response)
- Exposure duration
- 24 hours
- Method
- Luciferase normalized to protein; estradiol reference agonist
- Reported limits
- Exact EC50 and dose series not transcribed because supplementary numeric tables were not retrieved. Vehicle not specified for this panel in the retrieved assay description.
Methods and source locations
Compound reported in the paper: Levonorgestrel; Compound label in the paper: LNG
Reported levonorgestrel material and experimental system; no verification of Adooq A10529 or lot equivalence.
Sections 2.1–2.6 and 3.2; Figure 2G–I and caption
ERα-mediated reporter transactivation: Estradiol maximum set as 100%; pooled independent experiments in triplicate; at least two independent experiments in general methods; SEM — Sections 2.1–2.6 and 3.2; Figure 2G–I and caption
Mammary organoid transcription and AR-antagonist control
Cell-based study · Ex vivo C57Bl/6 mouse mammary organoids [3]
LNG increased Rankl and Wnt4 transcripts. Enzalutamide at 100 µM reduced LNG-induced Rankl expression, but did not significantly affect Wnt4 induction; 10 µM showed a nonsignificant trend for Rankl.
Literature-Reported Treatment Conditions
- Treatment concentration
- 20 nM LNG; antagonist-control experiment with 10 or 100 µM enzalutamide
- Exposure duration
- 6 hours
- Method
- RT-PCR transcript readout normalized to 36B4
- Vehicle
- Ethanol control; final concentration not specified in the read passage
- Co-treatment
- LNG alone for induction; LNG plus enzalutamide for AR-antagonist control. Combination effects are attributed separately.
- Reported limits
- Do not transfer BalbC time-course fold changes into this C57Bl/6 record. High antagonist concentration and n=3 limit inference.
Methods and source locations
Compound reported in the paper: Levonorgestrel; Compound label in the paper: LNG
Reported levonorgestrel material and experimental system; no verification of Adooq A10529 or lot equivalence.
Results: Effect of progestins on Rankl and Wnt4 transcript levels; AR activity is required for Rankl induction; Figure 1F,I and caption; Methods: Mouse tissue processing and hormone stimulation
Rankl and Wnt4 mRNA induction relative to vehicle: RT-PCR normalized to 36B4; caption n=6 for LNG; means ± SD — Figure 1F and associated Results paragraph
Effect of enzalutamide on LNG-induced transcription: n=3 independent experiments; means ± SD; caption specifies random-intercept linear model — Figure 1I and Results: AR activity is required for Rankl induction
