Nutr Res Pract.  2014 Oct;8(5):487-493. 10.4162/nrp.2014.8.5.487.

D. candidum has in vitro anticancer effects in HCT-116 cancer cells and exerts in vivo anti-metastatic effects in mice

Affiliations
  • 1College of Food Science, Southwest University, 2 Tiansheng Road, Beibei District, Chongqing 400067, China. birget@swu.edu.cn
  • 2Department of Biological and Chemical Engineering, Chongqing University of Education, China.

Abstract

BACKGROUND/OBJECTIVES
D. candidum is a traditional Chinese food or medicine widely used in Asia. There has been little research into the anticancer effects of D. candidum, particularly the effects in colon cancer cells. The aim of this study was to investigate the anticancer effects of D. candidum in vitro and in vivo.
MATERIALS/METHODS
The in vitro anti-cancer effects on HCT-116 colon cancer cells and in vivo anti-metastatic effects of DCME (Dendrobium canidum methanolic extract) were examined using the experimental methods of MTT assay, DAPI staining, flow cytometry analysis, RT-PCR, and Western blot analysis.
RESULTS
At a concentration of 1.0 mg/mL, DCME inhibited the growth of HCT-116 cells by 84%, which was higher than at concentrations of 0.5 and 0.25 mg/mL. Chromatin condensation and formation of apoptotic bodies were observed in cancer cells cultured with DCME as well. In addition, DCME induced significant apoptosis in cancer cells by upregulation of Bax, caspase 9, and caspase 3, and downregulation of Bcl-2. Expression of genes commonly associated with inflammation, NF-kappaB, iNOS, and COX-2, was significantly downregulated by DCME. DCME also exerted an anti-metastasis effect on cancer cells as demonstrated by decreased expression of MMP genes and increased expression of TIMPs, which was confirmed by the inhibition of induced tumor metastasis in colon 26-M3.1 cells in BALB/c mice.
CONCLUSIONS
Our results demonstrated that D. candidum had a potent in vitro anti-cancer effect, induced apoptosis, exhibited anti-inflammatory activities, and exerted in vivo anti-metastatic effects.

Keyword

D. candidum; HCT-116 human colon carcinoma cells; anti-cancer; apoptosis; anti-metastasis

MeSH Terms

Animals
Apoptosis
Asia
Asian Continental Ancestry Group
Blotting, Western
Caspase 3
Caspase 9
Chromatin
Colon
Colonic Neoplasms
Down-Regulation
Flow Cytometry
HCT116 Cells
Humans
Inflammation
Methanol
Mice*
Neoplasm Metastasis
NF-kappa B
Up-Regulation
Caspase 3
Caspase 9
Chromatin
Methanol
NF-kappa B

Figure

  • Fig. 1 Exposure of HCT-116 human colon cancer cells to DCME induces apoptosis. (A) Appearance of apoptotic bodies in HCT-116 cells treated with different concentrations of DCME for 48 h. (B) Treatment with DCME for 48 h increased the number of apoptotic cells as measured by flow cytometry. The profile represents an increased sub-G1 population (apoptotic cells) and each point represents the mean ± SD of three independent experiments.

  • Fig. 2 Effects of DCME on the mRNA and protein expressions of Bax, Bcl-2, and caspases in HCT-116 human colon cancer cells. (A) RT-PCR; (B) Western blot.

  • Fig. 3 Effects of DCME on the mRNA and protein expressions of NF-κB and IκB-α in HCT-116 human colon cancer cells. (A) RT-PCR; (B) Western blot.

  • Fig. 4 Effects of DCME on the mRNA and protein expressions of MMPs and TIMPs in HCT-116 human colon cancer cells. (A) RT-PCR; (B) Western blot.


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