1α,25-Dihydroxyvitamin D3 synergistically enhances anticancer effects of ginsenoside Rh2 in human prostate cancer cells

• Published in: The Journal of steroid biochemistry and molecular biology 2021-05, Vol.209, p.105828-105828, Article 105828
• Main Authors: Ben-Eltriki, Mohamed, Deb, Subrata, Guns, Emma S. Tomlinson
• Format: Article
• Language: English
• Subjects: 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3); Androgen receptors; Androgens; Antitumor activity; Apoptosis; BAX protein; Bcl-2 protein; Calcitriol; Caspase-3; Cell growth; Cell proliferation; Cell viability; Ginsenoside Rh2; Ginsenosides; Nuclear receptors; Prostate cancer; Protein expression; Synergism; Toxicity; Vitamin D; Vitamin D receptor; Vitamin D receptors; Vitamin D3
• ISSN: 0960-0760; 1879-1220
• DOI: 10.1016/j.jsbmb.2021.105828

•1α,25-dihydroxyvitamin D3 (1,25(OH)2D3) and ginsenoside Rh2 have anticancer effects as a single agent.•The combination of 1,25(OH)2D3 and Rh2 synergistically decreases cell viability and proliferation.•1,25(OH)2D3 and Rh2 differentially regulate androgen receptor and vitamin D receptor.•Potential synergistic mechanisms are demonstrated by 1,25(OH)2D3 and Rh2 through modulation of apoptotic proteins.•Anticancer effects of 1,25(OH)2D3 may be achieved at a lower condcentration when combined with Rh2. 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3, commonly known as calcitriol), the most active metabolite of vitamin D3, and ginsenoside Rh2 can regulate cellular differentiation and proliferation proteins. The purpose of the present study was to assess the effect of 1,25(OH)2D3 on the anticancer activities of Rh2 in human prostate cancer cells such as androgen-dependent LNCaP and androgen-independent C4-2 in vitro. The effects of treatment with 1,25(OH)2D3 or Rh2, either alone or in combination, on prostate cancer cells were evaluated through tetrazolium-based cell viability assay, BrdU cell proliferation rate estimation assay, and Western blot protein expression analyses of nuclear receptors (androgen receptor and vitamin D receptors) and apoptotic proteins (Bcl-2, Bax, and Caspase 3). The Combination Indices (CI) and Dose Reduction Indices (DRI) of 1,25(OH)2D3 and Rh2 were calculated to determine synergistic anticancer activity using Calcusyn software (Biosoft, Cambridge, UK). The cell viability assay data indicate that Rh2 treatment alone inhibited cell viability in a concentration-dependent manner and the addition of 10 nM 1,25(OH)2D3 to Rh2 significantly enhanced its ability to reduce cell viability up to 80 % in both the cell lines. Similarly, addition of 10 nM 1,25(OH)2D3 to Rh2 significantly lowered its IC50 values for cell proliferation from the range of 32–65 μM to 14-8 μM in LNCaP and C4-2 cells. In addition, protein expression analyses indicated that the combined treatment with Rh2 and 1,25(OH)2D3 led to greater downregulation of androgen receptor expression compared to single agent exposure. Similarly, the presence of 1,25(OH)2D3 synergistically increased the pro-apoptotic actions of Rh2 in both the cell lines. Overall, 1,25(OH)2D3 augments the Rh2-mediated anticancer effects through stimulating apoptosis and reduced cell proliferation which suggests that synergism of this combination may lead to potential lower need of the active vitamin D3 and limited to