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Ursolic acid from Trailliaedoxa gracilis induces apoptosis in medullary thyroid carcinoma cells.

Aguiriano-Moser V, Svejda B, Li ZX, Sturm S, Stuppner H, Ingolic E, Höger H, Siegl V, Meier-Allard N, Sadjak A, Pfragner R - Mol Med Rep (2015)

Bottom Line: Reverse transcription‑quantitative polymerase chain reaction of nuclear factor‑κB essential modifier (NEMO) was performed to delineate the role of the apoptotic pathway following treatment with UA.The observation of early‑onset activation of caspase 8 suggested that the responsible factor was linked to NEMO, an anti‑apoptotic protein.However, no differences in the mRNA transcription levels of NEMO were detected in MTC‑SK cells treated with UA, suggesting that this protein was not associated with the signal transducer and activator of transcription 3 pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathophysiology and Immunology, Center of Molecular Medicine, Medical University of Graz, Graz A‑8010, Austria.

ABSTRACT
Medullary thyroid carcinoma (MTC) originates from the C‑cells of the thyroid and is not sensitive to radiation or chemotherapy. Therefore, surgical removal of the tumor tissue in its entirety is the only curative treatment for MTC. The present study aimed to examine the potential mechanisms of action of extracts of Trailliaedoxa gracilis (TG; WW Smith & Forrest), a plant from the province of Sichuan, China, and of ursolic acid (UA), a pentacyclic triterpen present in TG, on the MTC‑SK MTC cell line. A total of 13 TG fractions and UA were examined in vitro for their effects on cell morphology, cell number, proliferation and rates of apoptosis. Reverse transcription‑quantitative polymerase chain reaction of nuclear factor‑κB essential modifier (NEMO) was performed to delineate the role of the apoptotic pathway following treatment with UA. TG and UA were examined in vivo in xenotransplanted MTC‑bearing severe combined immunodeficient mice. The TG fractions exhibited antiproliferative effects, with inhibition of mitochondrial activity in the tumor cells at concentrations, which caused no impairment of the normal control cells. The apoptotic rates of the MTC‑SK cells treated with the TG fractions and UA were determined, in which no marked tumor inhibition was observed in the treated MTC‑mice, and no change in the expression of NEMO was detected in the treated MTC‑SK cells. The observation of early‑onset activation of caspase 8 suggested that the responsible factor was linked to NEMO, an anti‑apoptotic protein. However, no differences in the mRNA transcription levels of NEMO were detected in MTC‑SK cells treated with UA, suggesting that this protein was not associated with the signal transducer and activator of transcription 3 pathway.

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Morphology of the MTC-SK cells treated with TG and UA. (A-D) Dissociation of cell aggregates following 24 h incubation (magnification, ×400). (B) 10 μg/ml TG-E5-F25 and (D) 10 μM UA. (E-H) Pro-apoptotic cell bodies and nuclei damage following 4 h incubation (magnification, ×400). (F) 10 μg/ml TG-E5-F25 and (H) 10 μM UA. (I-L) Structural damage and disintegration of clusters following 45 h incubation. (J and L) 25 μg/ml TG-E5-F25. (I and J) magnification, ×3,100; (K) magnification, ×13,300; (L) magnification, ×20,000. (A, C, E, G, I and K) Respective controls treated with dimethyl sulfoxide. UA, ursolic acid; TG, Trailliaedoxa gracilis.
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f3-mmr-12-04-5003: Morphology of the MTC-SK cells treated with TG and UA. (A-D) Dissociation of cell aggregates following 24 h incubation (magnification, ×400). (B) 10 μg/ml TG-E5-F25 and (D) 10 μM UA. (E-H) Pro-apoptotic cell bodies and nuclei damage following 4 h incubation (magnification, ×400). (F) 10 μg/ml TG-E5-F25 and (H) 10 μM UA. (I-L) Structural damage and disintegration of clusters following 45 h incubation. (J and L) 25 μg/ml TG-E5-F25. (I and J) magnification, ×3,100; (K) magnification, ×13,300; (L) magnification, ×20,000. (A, C, E, G, I and K) Respective controls treated with dimethyl sulfoxide. UA, ursolic acid; TG, Trailliaedoxa gracilis.

Mentions: The MTC-SK cells treated with the majority of the screened TG fractions and UA revealed altered morphology compared with the DMSO-treated control cells. A dissociation of cell aggregates was observed following incubation for 24 h (Fig. 3A–D). DAPI staining revealed that the MTC-SK cells treated with four of the fractions, particularly TG-E5-F25, exhibited pro-apoptotic cell bodies following incubation for 4 h (Fig. 3E and F), however, the majority of the fractions produced no specific pro-apoptotic cell bodies in the treated MTC-SK cells. The nuclei of the MTC-SK cells treated with either the TG fractions or UA were damaged compared with the DMSO-treated MTC-SK cells (Fig. 3G and H). Transmission electron microscopy revealed structural damage and disintegration of clusters in the MTC-SK cells treated with TG-E5-F28 compared with the DMSO-treated MTC-SK cells. The heterochromatin/euchromatin structure was reduced, cell organelles were damaged and pro-apoptotic vesicles were observed.


Ursolic acid from Trailliaedoxa gracilis induces apoptosis in medullary thyroid carcinoma cells.

Aguiriano-Moser V, Svejda B, Li ZX, Sturm S, Stuppner H, Ingolic E, Höger H, Siegl V, Meier-Allard N, Sadjak A, Pfragner R - Mol Med Rep (2015)

Morphology of the MTC-SK cells treated with TG and UA. (A-D) Dissociation of cell aggregates following 24 h incubation (magnification, ×400). (B) 10 μg/ml TG-E5-F25 and (D) 10 μM UA. (E-H) Pro-apoptotic cell bodies and nuclei damage following 4 h incubation (magnification, ×400). (F) 10 μg/ml TG-E5-F25 and (H) 10 μM UA. (I-L) Structural damage and disintegration of clusters following 45 h incubation. (J and L) 25 μg/ml TG-E5-F25. (I and J) magnification, ×3,100; (K) magnification, ×13,300; (L) magnification, ×20,000. (A, C, E, G, I and K) Respective controls treated with dimethyl sulfoxide. UA, ursolic acid; TG, Trailliaedoxa gracilis.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581794&req=5

f3-mmr-12-04-5003: Morphology of the MTC-SK cells treated with TG and UA. (A-D) Dissociation of cell aggregates following 24 h incubation (magnification, ×400). (B) 10 μg/ml TG-E5-F25 and (D) 10 μM UA. (E-H) Pro-apoptotic cell bodies and nuclei damage following 4 h incubation (magnification, ×400). (F) 10 μg/ml TG-E5-F25 and (H) 10 μM UA. (I-L) Structural damage and disintegration of clusters following 45 h incubation. (J and L) 25 μg/ml TG-E5-F25. (I and J) magnification, ×3,100; (K) magnification, ×13,300; (L) magnification, ×20,000. (A, C, E, G, I and K) Respective controls treated with dimethyl sulfoxide. UA, ursolic acid; TG, Trailliaedoxa gracilis.
Mentions: The MTC-SK cells treated with the majority of the screened TG fractions and UA revealed altered morphology compared with the DMSO-treated control cells. A dissociation of cell aggregates was observed following incubation for 24 h (Fig. 3A–D). DAPI staining revealed that the MTC-SK cells treated with four of the fractions, particularly TG-E5-F25, exhibited pro-apoptotic cell bodies following incubation for 4 h (Fig. 3E and F), however, the majority of the fractions produced no specific pro-apoptotic cell bodies in the treated MTC-SK cells. The nuclei of the MTC-SK cells treated with either the TG fractions or UA were damaged compared with the DMSO-treated MTC-SK cells (Fig. 3G and H). Transmission electron microscopy revealed structural damage and disintegration of clusters in the MTC-SK cells treated with TG-E5-F28 compared with the DMSO-treated MTC-SK cells. The heterochromatin/euchromatin structure was reduced, cell organelles were damaged and pro-apoptotic vesicles were observed.

Bottom Line: Reverse transcription‑quantitative polymerase chain reaction of nuclear factor‑κB essential modifier (NEMO) was performed to delineate the role of the apoptotic pathway following treatment with UA.The observation of early‑onset activation of caspase 8 suggested that the responsible factor was linked to NEMO, an anti‑apoptotic protein.However, no differences in the mRNA transcription levels of NEMO were detected in MTC‑SK cells treated with UA, suggesting that this protein was not associated with the signal transducer and activator of transcription 3 pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathophysiology and Immunology, Center of Molecular Medicine, Medical University of Graz, Graz A‑8010, Austria.

ABSTRACT
Medullary thyroid carcinoma (MTC) originates from the C‑cells of the thyroid and is not sensitive to radiation or chemotherapy. Therefore, surgical removal of the tumor tissue in its entirety is the only curative treatment for MTC. The present study aimed to examine the potential mechanisms of action of extracts of Trailliaedoxa gracilis (TG; WW Smith & Forrest), a plant from the province of Sichuan, China, and of ursolic acid (UA), a pentacyclic triterpen present in TG, on the MTC‑SK MTC cell line. A total of 13 TG fractions and UA were examined in vitro for their effects on cell morphology, cell number, proliferation and rates of apoptosis. Reverse transcription‑quantitative polymerase chain reaction of nuclear factor‑κB essential modifier (NEMO) was performed to delineate the role of the apoptotic pathway following treatment with UA. TG and UA were examined in vivo in xenotransplanted MTC‑bearing severe combined immunodeficient mice. The TG fractions exhibited antiproliferative effects, with inhibition of mitochondrial activity in the tumor cells at concentrations, which caused no impairment of the normal control cells. The apoptotic rates of the MTC‑SK cells treated with the TG fractions and UA were determined, in which no marked tumor inhibition was observed in the treated MTC‑mice, and no change in the expression of NEMO was detected in the treated MTC‑SK cells. The observation of early‑onset activation of caspase 8 suggested that the responsible factor was linked to NEMO, an anti‑apoptotic protein. However, no differences in the mRNA transcription levels of NEMO were detected in MTC‑SK cells treated with UA, suggesting that this protein was not associated with the signal transducer and activator of transcription 3 pathway.

Show MeSH
Related in: MedlinePlus