Ann Dermatol.  2016 Jun;28(3):327-334. 10.5021/ad.2016.28.3.327.

Epigallocatechin Gallate-Mediated Alteration of the MicroRNA Expression Profile in 5α-Dihydrotestosterone-Treated Human Dermal Papilla Cells

• Shin S
• Kim K
• Lee MJ
• Lee J
• Choi S
• Kim KS
• Ko JM
• Han H
• Kim SY
• Youn HJ
• Ahn KJ
• An IS
• An S
• Cha HJ

• KMID: 2164640
• DOI: http://doi.org/10.5021/ad.2016.28.3.327

Abstract

BACKGROUND
Dihydrotestosterone (DHT) induces androgenic alopecia by shortening the hair follicle growth phase, resulting in hair loss. We previously demonstrated how changes in the microRNA (miRNA) expression profile influenced DHT-mediated cell death, cell cycle arrest, cell viability, the generation of reactive oxygen species (ROS), and senescence. Protective effects against DHT have not, however, been elucidated at the genome level.
OBJECTIVE
We showed that epigallocatechin gallate (EGCG), a major component of green tea, protects DHT-induced cell death by regulating the cellular miRNA expression profile.
METHODS
We used a miRNA microarray to identify miRNA expression levels in human dermal papilla cells (DPCs). We investigated whether the miRNA expression influenced the protective effects of EGCG against DHT-induced cell death, growth arrest, intracellular ROS levels, and senescence.
RESULTS
EGCG protected against the effects of DHT by altering the miRNA expression profile in human DPCs. In addition, EGCG attenuated DHT-mediated cell death and growth arrest and decreased intracellular ROS levels and senescence. A bioinformatics analysis elucidated the relationship between the altered miRNA expression and EGCG-mediated protective effects against DHT.
CONCLUSION
Overall, our results suggest that EGCG ameliorates the negative effects of DHT by altering the miRNA expression profile in human DPCs.

Keyword

Epigallocatechin gallate; Human dermal papilla cells; MicroRNAs

MeSH Terms

Aging
Alopecia
Cell Cycle Checkpoints
Cell Death
Cell Survival
Computational Biology
Dihydrotestosterone
Genome
Hair
Hair Follicle
Humans*
MicroRNAs*
Reactive Oxygen Species
Tea
Dihydrotestosterone
MicroRNAs
Reactive Oxygen Species
Tea

Figure

• 

  Fig. 1 Effects of epigallocatechin gallate (EGCG) on human dermal papilla cell (DPC) viability and cell cycle progression. Human DPCs were treated with different concentrations of EGCG (A) or EGCG and dihydrotestosterone (DHT) (B) for 24 hours before water soluble tetrazolium salt (WST-1) assays to measure viability. (C) Human DPCs were treated with 10 µM EGCG or 10–4 M DHT for 24 hours before they were washed and stained with propidium iodide (PI; Sigma-Aldrich, St. Louis, MO, USA). The fluorescence intensity distributions of the stained cells were analyzed by flow cytometry (left) and the proportions of the cells in Sub-G1 phase were determined (right). Cells in the Sub-G1, G1, S, and G2/M phases were separated using the gates M1, M2, M3, and M4, respectively. Viability data are expressed as percentages of the viability of the internal control (untreated sample) and are shown as the mean±standard error of the mean of three independent experiments. #p<0.05 compared with vehicle, *p<0.05 compared with DHT.

• 

  Fig. 2 Effects of epigallocatechin gallate (EGCG) on intracellular reactive oxygen species (ROS) levels and cellular senescence in human dermal papilla cells (DPCs). (A) Human DPCs were grown with or without 10 µM EGCG and 10–4 M dihydrotestosterone (DHT). To detect intracellular ROS levels, we performed 2',7'-dichlorofluorescein diacetate staining and detected cell cycle changes using a FACScalibur flow cytometer (BD Biosciences, San Jose, CA, USA). Changes in intracellular ROS levels were determined with the M1 gate. (B) Cells were treated with 10 µM EGCG or 10–4 M DHT for 48 hours. Senescence was measured using senescence- associated β-gal assays. Senescence data are expressed as percentages of the senescence of the internal control (untreated sample) and are shown as the mean±standard error of the mean of three independent experiments. PI: propidium iodide (Sigma-Aldrich, St. Louis, MO, USA). #p<0.05 compared with vehicle, *p<0.05 compared with DHT.

• 

  Fig. 3 Gene ontology classification of predicted target genes of microRNAs affected by epigallocatechin gallate in dihydrotestosterone-treated human dermal papilla cells. GO terms 1~5 are related to anti-oxidation: 1-oxidoreduction coenzyme metabolic process (GO:0006733); 2-response to oxidative stress (GO:0006979); 3-response to hydrogen peroxide (GO:0042542); 4-energy derivation by oxidation of organic compounds (GO:0015980); 5-oxidation reduction (GO:0055114). GO terms 6~22 are related to apoptosis and cell death: 6-anti-apoptosis (GO:0006916); 7-induction of apoptosis by extracellular signals (GO:0008624); 8-regulation of apoptosis (GO:0042981); 9-negative regulation of apoptosis (GO:0043066); 10-positive regulation of apoptosis (GO:0043065); 11-induction of apoptosis (GO:0006917); 12-apoptosis (GO:0006915); 13-regulation of programmed cell death (GO:0043067); 14-regulation of cell death (GO:0010941); 15-negative regulation of programmed cell death (GO:0043069); 16-negative regulation of cell death (GO:0060548); 17-positive regulation of programmed cell death (GO:0043068); 18-positive regulation of cell death (GO:0010942); 19-induction of programmed cell death (GO:0012502); 20-cell death (GO:0008219); 21-death (GO:0016265); 22-programmed cell death (GO:0012501). GO terms 23~33 are related to proliferation and cell growth: 23-cell proliferation (GO:0008283); 24-positive regulation of cell proliferation (GO:0008284); 25-regulation of cell proliferation (GO:0042127); 26-negative regulation of cell proliferation (GO:0008285); 27-negative regulation of cell growth (GO:0030308); 28-regulation of growth (GO:0040008); 29-regulation of developmental growth (GO:0048638); 30-negative regulation of growth (GO:0045926); 31-regulation of cell growth (GO:0001558); 32-positive regulation of growth (GO:0045927); 33-growth (GO:0040007). GO term 34 is related to aging (34-aging (GO:0007568). GO terms 35~48 are related to the cell cycle: 35-regulation of cell cycle (GO:0051726); 36-negative regulation of cell cycle (GO:0045786); 37-M phase of meiotic cell cycle (GO:0051327); 38-meiotic cell cycle (GO:0051321); 39-negative regulation of mitotic cell cycle (GO:0045930); 40-negative regulation of cell cycle process (GO:0010948); 41-regulation of mitotic cell cycle (GO:0007346); 42-cell cycle phase (GO:0022403); 43-regulation of cell cycle process (GO:0010564); 44-cell cycle (GO:0007049); 45-cell cycle process (GO:0022402); 46-mitotic cell cycle (GO:0000278); 47-cell cycle checkpoint (GO:0000075); 48-M phase of mitotic cell cycle (GO:0000087).

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