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. 2015 Jan;56(1):16-23.
doi: 10.3349/ymj.2015.56.1.16.

Anthocyanin induces apoptosis of DU-145 cells in vitro and inhibits xenograft growth of prostate cancer

U Syn Ha  1 , Woong Jin Bae  2 , Su Jin Kim  1 , Byung Il Yoon  1 , Sung Hoo Hong  1 , Ji Youl Lee  1 , Tae Kon Hwang  1 , Sung Yeoun Hwang  3 , Zhiping Wang  4 , Sae Woong Kim  5
Affiliations

Anthocyanin induces apoptosis of DU-145 cells in vitro and inhibits xenograft growth of prostate cancer

U Syn Ha et al. Yonsei Med J. 2015 Jan.

Abstract

Purpose: To investigate the effects of anthocyanins extracted from black soybean, which have antioxidant activity, on apoptosis in vitro (in hormone refractory prostate cancer cells) and on tumor growth in vivo (in athymic nude mouse xenograft model).

Materials and methods: The growth and viability of DU-145 cells treated with anthocyanins were assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and apoptosis was assessed by DNA laddering. Immunoblotting was conducted to evaluate differences in the expressions of p53, Bax, Bcl, androgen receptor (AR), and prostate specific antigen (PSA). To study the inhibitory effects of anthocyanins on tumor growth in vivo, DU-145 tumor xenografts were established in athymic nude mice. The anthocyanin group was treated with daily oral anthocyanin (8 mg/kg) for 14 weeks. After 2 weeks of treatment, DU-145 cells (2×10⁶) were inoculated subcutaneously into the right flank to establish tumor xenografts. Tumor dimensions were measured twice a week using calipers and volumes were calculated.

Results: Anthocyanin treatment of DU-145 cells resulted in 1) significant increase in apoptosis in a dose-dependent manner, 2) significant decrease in p53 and Bcl-2 expressions (with increased Bax expression), and 3) significant decrease in PSA and AR expressions. In the xenograft model, anthocyanin treatment significantly inhibit tumor growth.

Conclusion: This study suggests that anthocyanins from black soybean inhibit the progression of prostate cancer in vitro and in a xenograft model.

Keywords: Prostatic neoplasms; anthocyanins; apoptosis.

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Conflict of interest statement

The authors have no financial conflicts of interest.

Figures

Fig. 1
Fig. 1
Effect of anthocyanin on the viability and apoptosis of DU-145 cells. (A) DU-145 cells were treated with specified anthocyanin concentrations at each treatment time, and cell viability was determined by MTT assay. (B) Viability of DU-145 cells in specified anthocyanin concentrations after 24 hr. (C) Results of a DNA laddering assay in DU-145 cells treated with specified anthocyanin concentrations.
Fig. 2
Fig. 2
The effect of anthocyanin on p53, Bax, and Bcl-2 Expression in DU-145 cells. (A) Western blot analysis of p53, Bax, and Bcl-2. (B) Densitometric analysis of Bax relative to β-actin. (C) Densitometric analysis of Bcl-2 relative to β-actin. (D) The relative ratio of Bax to Bcl-2. (E) Densitometric analysis of p53 relative to β-actin.
Fig. 3
Fig. 3
The effect of anthocyanin on PSA and androgen receptor expression in DU-145 cells. (A) Western blot analysis of PSA and AR. (B) Densitometric analysis of PSA relative to β-actin. (C) Densitometric analysis of AR relative to β-actin. PSA, prostate specific antigen; AR, androgen receptor.
Fig. 4
Fig. 4
The effect of anthocyanin on the ratio of NAD+ to NADH in DU-145 cells. NAD+, nicotinamide adenine dinucleotide.
Fig. 5
Fig. 5
The effect of anthocyanin on tumor growth.
See this image and copyright information in PMC

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