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The in vitro and in vivo anti-cancer activities of a standardized quassinoids composition from Eurycoma longifolia on LNCaP human prostate cancer cells.

Tong KL, Chan KL, AbuBakar S, Low BS, Ma HQ, Wong PF - PLoS ONE (2015)

Bottom Line: Quassinoids are a group of diterpenoids found in plants from the Simaroubaceae family.Therefore, the present study investigates the effects of a standardized total quassinoids composition (SQ40) containing 40% of the total quassinoids found in E. longifolia on LNCaP human prostate cancer cell line.Moreover, SQ40 also inhibited androgen receptor translocation to nucleus which is important for the transactivation of its target gene, prostate-specific antigen (PSA) and resulted in a significant reduction of PSA secretion after the treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.

ABSTRACT
Quassinoids are a group of diterpenoids found in plants from the Simaroubaceae family. They are also the major bioactive compounds found in Eurycoma longifolia which is commonly used as traditional medicine in South East Asia to treat various ailments including sexual dysfunction and infertility. These uses are attributed to its ability to improve testosterone level in men. Chronic consumption of E. longifolia extracts has been reported to increase testosterone level in men and animal model but its effect on prostate growth remains unknown. Therefore, the present study investigates the effects of a standardized total quassinoids composition (SQ40) containing 40% of the total quassinoids found in E. longifolia on LNCaP human prostate cancer cell line. SQ40 inhibited LNCaP cell growth at IC50 value of 5.97 μg/mL while the IC50 on RWPE-1 human prostate normal cells was 59.26 μg/mL. SQ40 also inhibited 5α-dihydrotestosterone-stimulated growth in LNCaP cells dose-dependently. The inhibitory effect of SQ40 in anchorage-independent growth of LNCaP cells was also demonstrated using soft agar assay. SQ40 suppressed LNCaP cell growth via G0/G1 phase arrest which was accompanied by the down-regulation of CDK4, CDK2, Cyclin D1 and Cyclin D3 and up-regulation of p21Waf1/Cip1 protein levels. SQ40 at higher concentrations or longer treatment duration can cause G2M growth arrest leading to apoptotic cell death as demonstrated by the detection of poly(ADP-ribose) polymerase cleavage in LNCaP cells. Moreover, SQ40 also inhibited androgen receptor translocation to nucleus which is important for the transactivation of its target gene, prostate-specific antigen (PSA) and resulted in a significant reduction of PSA secretion after the treatment. In addition, intraperitoneal injection of 5 and 10 mg/kg of SQ40 also significantly suppressed the LNCaP tumor growth on mouse xenograft model. Results from the present study suggest that the standardized total quassinoids composition from E. longifolia promotes anti-prostate cancer activities in LNCaP human prostate cancer cells.

No MeSH data available.


Related in: MedlinePlus

Quantitative measurement of the level of nuclear AR and PSA secretion in SQ40-treated LNCaP cells.LNCaP cells were first cultured in growth media supplemented with 5% CSS for 48 hours and then treated with 3, 6 and 12 μg/mL of SQ40 in the presence or absence of 100 nM DHT for 72 hours. Nuclear fraction of AR obtained from the cell lysate and concentrations of PSA secreted into culture medium were measured using commercial available ELISA kit. (A) AR level was normalized to the total nuclear protein level while (B) the concentration of PSA were normalised to the total cell number. Data were expressed as means ± SEM of three independent experiments and indicated as percentage of 100 nM DHT-stimulated cells set at 100%. ** indicates p<0.01; ***, p<0.001 versus unstimulated control. ###, p<0.001 versus 100 nM DHT-stimulated cells.
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pone.0121752.g007: Quantitative measurement of the level of nuclear AR and PSA secretion in SQ40-treated LNCaP cells.LNCaP cells were first cultured in growth media supplemented with 5% CSS for 48 hours and then treated with 3, 6 and 12 μg/mL of SQ40 in the presence or absence of 100 nM DHT for 72 hours. Nuclear fraction of AR obtained from the cell lysate and concentrations of PSA secreted into culture medium were measured using commercial available ELISA kit. (A) AR level was normalized to the total nuclear protein level while (B) the concentration of PSA were normalised to the total cell number. Data were expressed as means ± SEM of three independent experiments and indicated as percentage of 100 nM DHT-stimulated cells set at 100%. ** indicates p<0.01; ***, p<0.001 versus unstimulated control. ###, p<0.001 versus 100 nM DHT-stimulated cells.

Mentions: The effect of SQ40 on the AR protein of LNCaP was investigated next. Nuclear AR extract in 100 nM DHT stimulated-LNCaP cells increased by 25% compared to the unstimulated cells (Fig. 7A). SQ40 treatment at 3 and 6 μg/mL alone decreased the basal nuclear AR level by 25% as well when compared to the unstimulated cells (Fig. 7A; p>0.05). Therefore, combination treatment of DHT and the quassinoids composition at 3 and 6 μg/mL did not significantly affect the nuclear AR level when compared to the unstimulated cells (Fig. 7A; p>0.05). However, treatment with 12 μg/mL of SQ40 significantly suppressed the basal nuclear AR level and its suppression could not be overcome by the presence of DHT (Fig. 7A; p<0.001).


The in vitro and in vivo anti-cancer activities of a standardized quassinoids composition from Eurycoma longifolia on LNCaP human prostate cancer cells.

Tong KL, Chan KL, AbuBakar S, Low BS, Ma HQ, Wong PF - PLoS ONE (2015)

Quantitative measurement of the level of nuclear AR and PSA secretion in SQ40-treated LNCaP cells.LNCaP cells were first cultured in growth media supplemented with 5% CSS for 48 hours and then treated with 3, 6 and 12 μg/mL of SQ40 in the presence or absence of 100 nM DHT for 72 hours. Nuclear fraction of AR obtained from the cell lysate and concentrations of PSA secreted into culture medium were measured using commercial available ELISA kit. (A) AR level was normalized to the total nuclear protein level while (B) the concentration of PSA were normalised to the total cell number. Data were expressed as means ± SEM of three independent experiments and indicated as percentage of 100 nM DHT-stimulated cells set at 100%. ** indicates p<0.01; ***, p<0.001 versus unstimulated control. ###, p<0.001 versus 100 nM DHT-stimulated cells.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4380335&req=5

pone.0121752.g007: Quantitative measurement of the level of nuclear AR and PSA secretion in SQ40-treated LNCaP cells.LNCaP cells were first cultured in growth media supplemented with 5% CSS for 48 hours and then treated with 3, 6 and 12 μg/mL of SQ40 in the presence or absence of 100 nM DHT for 72 hours. Nuclear fraction of AR obtained from the cell lysate and concentrations of PSA secreted into culture medium were measured using commercial available ELISA kit. (A) AR level was normalized to the total nuclear protein level while (B) the concentration of PSA were normalised to the total cell number. Data were expressed as means ± SEM of three independent experiments and indicated as percentage of 100 nM DHT-stimulated cells set at 100%. ** indicates p<0.01; ***, p<0.001 versus unstimulated control. ###, p<0.001 versus 100 nM DHT-stimulated cells.
Mentions: The effect of SQ40 on the AR protein of LNCaP was investigated next. Nuclear AR extract in 100 nM DHT stimulated-LNCaP cells increased by 25% compared to the unstimulated cells (Fig. 7A). SQ40 treatment at 3 and 6 μg/mL alone decreased the basal nuclear AR level by 25% as well when compared to the unstimulated cells (Fig. 7A; p>0.05). Therefore, combination treatment of DHT and the quassinoids composition at 3 and 6 μg/mL did not significantly affect the nuclear AR level when compared to the unstimulated cells (Fig. 7A; p>0.05). However, treatment with 12 μg/mL of SQ40 significantly suppressed the basal nuclear AR level and its suppression could not be overcome by the presence of DHT (Fig. 7A; p<0.001).

Bottom Line: Quassinoids are a group of diterpenoids found in plants from the Simaroubaceae family.Therefore, the present study investigates the effects of a standardized total quassinoids composition (SQ40) containing 40% of the total quassinoids found in E. longifolia on LNCaP human prostate cancer cell line.Moreover, SQ40 also inhibited androgen receptor translocation to nucleus which is important for the transactivation of its target gene, prostate-specific antigen (PSA) and resulted in a significant reduction of PSA secretion after the treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.

ABSTRACT
Quassinoids are a group of diterpenoids found in plants from the Simaroubaceae family. They are also the major bioactive compounds found in Eurycoma longifolia which is commonly used as traditional medicine in South East Asia to treat various ailments including sexual dysfunction and infertility. These uses are attributed to its ability to improve testosterone level in men. Chronic consumption of E. longifolia extracts has been reported to increase testosterone level in men and animal model but its effect on prostate growth remains unknown. Therefore, the present study investigates the effects of a standardized total quassinoids composition (SQ40) containing 40% of the total quassinoids found in E. longifolia on LNCaP human prostate cancer cell line. SQ40 inhibited LNCaP cell growth at IC50 value of 5.97 μg/mL while the IC50 on RWPE-1 human prostate normal cells was 59.26 μg/mL. SQ40 also inhibited 5α-dihydrotestosterone-stimulated growth in LNCaP cells dose-dependently. The inhibitory effect of SQ40 in anchorage-independent growth of LNCaP cells was also demonstrated using soft agar assay. SQ40 suppressed LNCaP cell growth via G0/G1 phase arrest which was accompanied by the down-regulation of CDK4, CDK2, Cyclin D1 and Cyclin D3 and up-regulation of p21Waf1/Cip1 protein levels. SQ40 at higher concentrations or longer treatment duration can cause G2M growth arrest leading to apoptotic cell death as demonstrated by the detection of poly(ADP-ribose) polymerase cleavage in LNCaP cells. Moreover, SQ40 also inhibited androgen receptor translocation to nucleus which is important for the transactivation of its target gene, prostate-specific antigen (PSA) and resulted in a significant reduction of PSA secretion after the treatment. In addition, intraperitoneal injection of 5 and 10 mg/kg of SQ40 also significantly suppressed the LNCaP tumor growth on mouse xenograft model. Results from the present study suggest that the standardized total quassinoids composition from E. longifolia promotes anti-prostate cancer activities in LNCaP human prostate cancer cells.

No MeSH data available.


Related in: MedlinePlus