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Preventive effects of Dendrobium hybrid extract on free radical activity and melanoma cell proliferation by inducing apoptotic gene expression

  • Authors:
    • Rungpailin Khonsap
    • Pilairath Meksangsee
    • Pichamon Kiatwuthinon
    • Nattanan Panjaworayan T‑Thienprasert
    • Napachanok Mongkoldhumrongkul Swainson
    • Wannarat Phonphoem
    • Pakorn Wattana‑Amorn
    • Attawan Aramrak
  • View Affiliations / Copyright

    Affiliations: Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand, Department of Chemistry, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand
    Copyright: © Khonsap et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 21
    |
    Published online on: November 27, 2025
       https://doi.org/10.3892/br.2025.2094
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Abstract

Dendrobium species are used in folk medicines due to their medicinal properties by acting as antioxidants and anticancer agents. Derived from parent plants used in dermatological treatments and cosmetics, Dendrobium Pearl Vera hybrid (DH) was extracted and investigated for its inhibitory effects on free radicals and skin cancer cell proliferation. Based on colorimetric spectrophotometry, the propanolic extract (DH‑P) from the whole plant contained flavonoids, phenolics and alkaloids at 80.11±0.69, 50.51±0.44 and 38.75±1.77 µg/g fresh weight, respectively. Using high‑resolution mass spectrometry, eight compounds involved in skin protection were annotated: Fuanosine, L‑glutamine, threonic acid, quercetagetin, shikimic acid, mannose, p‑coumaric acid and L‑malic acid. DH‑P scavenged free radicals in a concentration-dependent manner and inhibited the activity of collagenase and hyaluronidase, but not in a concentration‑dependent manner. Treatment using DH‑P for 96 h significantly decreased the proliferation of SK‑MEL‑28 melanoma cells in a dose‑dependent manner, with a half‑maximal inhibitory concentration of 20.23±1.04 µg/ml. Thus, DH‑P at 20 µg/ml was used in further studies. Cell migration was not inhibited by DH‑P. However, DH‑P may induce SK‑MEL‑28 cell death through apoptosis, as nuclear condensation and fragmentation was observed after 48 h DH‑P treatment. Gene expression analysis was performed to evaluate key signaling molecules involved in cell apoptosis, such as bax, bcl‑2, cytochrome c, caspases (cas)‑3 and ‑9 and p53. The expression levels of cas‑3 and cas‑9, executors in the apoptosis pathway, were significantly upregulated in the SK‑MEL‑28 cells treated with DH‑P for 48 h, further confirming that cell death may occur through an apoptosis mechanism.
View Figures

Figure 1

Determination of secondary
metabolites in DH-P by thin layer chromatography. (A) Chromatogram
of phenolic and flavonoid compounds in DH-P (10 mg/ml). (B) Amounts
of total phenolics, flavonoids and alkaloids in DH-P. n=6. DH-P,
propanolic extract from Dendrobium Pearl Vera hybrid; Q,
quercetin; FW, fresh weight.

Figure 2

Structures of tentative compounds
identified in propanolic extract from Dendrobium Pearl Vera
hybrid. (A) Guanosine. (B) L-glutamine. (C) Threonic acid. (D)
Quercetagetin. (E) Shikimic acid. (F) D-(+)-mannose. (G)
p-coumaric acid. (H) L-malic acid.

Figure 3

Scavenging effect of DH-P on free
radicals at various concentrations. Radical scavenging capacity of
DH-P for (A) DPPH and (B) ABTS radicals. (C) Reducing power on
ferric ions (Fe3+). n=6. DH-P, propanolic extract from
Dendrobium Pearl Vera hybrid; DPPH,
2,2-diphenyl-1-picrylhydrazyl; ABTS,
2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid); VCEAC,
vitamin C equivalent antioxidant capacity.

Figure 4

Effect of DH-P on extracellular
matrix enzyme activity at various concentrations. Inhibitory
effects of DH-P on (A) collagenase and (B) hyaluronidase enzyme
activity. Inhibitor controls were 1 M 1,10-PN and 1 mg/ml OA. n=6.
*P<0.05; **P<0.01;
****P<0.0001 vs. 800 or 25 µg/ml DH-P. OA, oleanolic
acid; DH-P, propanolic extract from Dendrobium Pearl Vera
hybrid; 1,10-PN, 1,10-phenanthroline.

Figure 5

Inhibitory effects of DH-P on cell
viability and migration. (A) IC50 value determined from
dose-response cytotoxicity analysis of DH-P (0-100 µg/ml). (B)
Quantitative analysis of SK-MEL-28 cell migration following
treatment with 20 µg/ml DH-P or 5% DMSO. n=6.
*P<0.05; **P<0.01; vs. DMSO. (C)
Transwell migration assay of SK-MEL-28 cells treated with DH-P for
72 h (n=4). DH-P, propanolic extract from Dendrobium Pearl
Vera hybrid; IC50, half-maximal inhibitory
concentration.

Figure 6

Nuclear morphology of SK-MEL-28 cells
treated with DH-P at 20 µg/ml for 0, 48, and 96 h. Nuclei were
stained using Hoechst 33342 (magnification, x40). Scale bar, 20 µm.
Green arrow, nuclei of healthy cells. Red arrow, condensed or
fragmented nuclei of apoptotic cells. DH-P, propanolic extract from
Dendrobium Pearl Vera hybrid.

Figure 7

Expression of apoptotic-related genes
in SK-MEL-28. Fold-change in cells treated with DH-P at 20 µg/ml
for 48 h were calculated relative to control group. (A)
cytc. (B) cas-3. (C) cas-9. (D) p53.
(E) bax. (F) bcl-2. n=6. **P<0.01;
****P<0.0001 vs. control. DH-P, propanolic extract
from Dendrobium Pearl Vera hybrid; cytc, cytochrome
c; cas, caspase.
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Spandidos Publications style
Khonsap R, Meksangsee P, Kiatwuthinon P, T‑Thienprasert NP, Swainson NM, Phonphoem W, Wattana‑Amorn P and Aramrak A: Preventive effects of <em>Dendrobium</em> hybrid extract on free radical activity and melanoma cell proliferation by inducing apoptotic gene expression. Biomed Rep 24: 21, 2026.
APA
Khonsap, R., Meksangsee, P., Kiatwuthinon, P., T‑Thienprasert, N.P., Swainson, N.M., Phonphoem, W. ... Aramrak, A. (2026). Preventive effects of <em>Dendrobium</em> hybrid extract on free radical activity and melanoma cell proliferation by inducing apoptotic gene expression. Biomedical Reports, 24, 21. https://doi.org/10.3892/br.2025.2094
MLA
Khonsap, R., Meksangsee, P., Kiatwuthinon, P., T‑Thienprasert, N. P., Swainson, N. M., Phonphoem, W., Wattana‑Amorn, P., Aramrak, A."Preventive effects of <em>Dendrobium</em> hybrid extract on free radical activity and melanoma cell proliferation by inducing apoptotic gene expression". Biomedical Reports 24.2 (2026): 21.
Chicago
Khonsap, R., Meksangsee, P., Kiatwuthinon, P., T‑Thienprasert, N. P., Swainson, N. M., Phonphoem, W., Wattana‑Amorn, P., Aramrak, A."Preventive effects of <em>Dendrobium</em> hybrid extract on free radical activity and melanoma cell proliferation by inducing apoptotic gene expression". Biomedical Reports 24, no. 2 (2026): 21. https://doi.org/10.3892/br.2025.2094
Copy and paste a formatted citation
x
Spandidos Publications style
Khonsap R, Meksangsee P, Kiatwuthinon P, T‑Thienprasert NP, Swainson NM, Phonphoem W, Wattana‑Amorn P and Aramrak A: Preventive effects of <em>Dendrobium</em> hybrid extract on free radical activity and melanoma cell proliferation by inducing apoptotic gene expression. Biomed Rep 24: 21, 2026.
APA
Khonsap, R., Meksangsee, P., Kiatwuthinon, P., T‑Thienprasert, N.P., Swainson, N.M., Phonphoem, W. ... Aramrak, A. (2026). Preventive effects of <em>Dendrobium</em> hybrid extract on free radical activity and melanoma cell proliferation by inducing apoptotic gene expression. Biomedical Reports, 24, 21. https://doi.org/10.3892/br.2025.2094
MLA
Khonsap, R., Meksangsee, P., Kiatwuthinon, P., T‑Thienprasert, N. P., Swainson, N. M., Phonphoem, W., Wattana‑Amorn, P., Aramrak, A."Preventive effects of <em>Dendrobium</em> hybrid extract on free radical activity and melanoma cell proliferation by inducing apoptotic gene expression". Biomedical Reports 24.2 (2026): 21.
Chicago
Khonsap, R., Meksangsee, P., Kiatwuthinon, P., T‑Thienprasert, N. P., Swainson, N. M., Phonphoem, W., Wattana‑Amorn, P., Aramrak, A."Preventive effects of <em>Dendrobium</em> hybrid extract on free radical activity and melanoma cell proliferation by inducing apoptotic gene expression". Biomedical Reports 24, no. 2 (2026): 21. https://doi.org/10.3892/br.2025.2094
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