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Andrographolide inhibits hypoxia-inducible factor-1 through phosphatidylinositol 3-kinase/AKT pathway and suppresses breast cancer growth.

Li J, Zhang C, Jiang H, Cheng J - Onco Targets Ther (2015)

Bottom Line: Further, Andro inhibited T47D and MDA-MB-231 cell proliferation and colony formation.In addition, it exhibited significant in vivo efficacy and antitumor potential against the MDA-MB-231 xenograft in nude mice.In conclusion, these results highlighted the potential effects of Andro, which inhibits HIF-1, and hence may be developed as an antitumor agent for breast cancer therapy in future.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery, Beijing Chao-Yang Hospital, Beijing, People's Republic of China.

ABSTRACT
Hypoxia-inducible factor-1 (HIF-1) is a master regulator of the transcriptional response to hypoxia. HIF-1α is one of the most compelling anticancer targets. Andrographolide (Andro) was newly identified to inhibit HIF-1 in T47D cells (a half maximal effective concentration [EC50] of 1.03×10(-7) mol/L), by a dual-luciferase reporter assay. It suppressed HIF-1α protein and gene accumulation, which was dependent on the inhibition of upstream phosphatidylinositol 3-kinase (PI3K)/AKT pathway. It also abrogated the expression of HIF-1 target vascular endothelial growth factor (VEGF) gene and protein. Further, Andro inhibited T47D and MDA-MB-231 cell proliferation and colony formation. In addition, it exhibited significant in vivo efficacy and antitumor potential against the MDA-MB-231 xenograft in nude mice. In conclusion, these results highlighted the potential effects of Andro, which inhibits HIF-1, and hence may be developed as an antitumor agent for breast cancer therapy in future.

No MeSH data available.


Related in: MedlinePlus

Andro specifically inhibited HIF-1α protein and gene accumulation under hypoxia.Notes: T47D and MDA-MB-231 cells were treated with or without Andro under hypoxia for 8 hours. The protein extracts from these cells were subjected to Western blotting for HIF-1α, HIF-2α, and HIF-1β expression. β-Actin was used as a loading control. Representative Western blotting images of T47D cells (A) and MDA-MB-231 cells (B) are shown. The quantitative data (mean ± SD) of T47D cells (C) and MDA-MB-231 (D) cells were calculated from at least three separate experiments, taking the ratio of HIF-1α, HIF-2α, and HIF-1β bands to the β-actin band in 1% O2 group. *P<0.05 and **P<0.01 compared with hypoxia group (1% O2). (E) Total RNAs were extracted and HIF-1α mRNA levels were analyzed by qPCR. *P<0.05 and **P<0.01 compared with hypoxia group (1% O2).Abbreviations: Andro, Andrographolide; HIF-1, hypoxia-inducible factor-1; SD, standard deviation; mRNA, messenger RNA; qPCR, real-time polymerase chain reaction.
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f2-ott-8-427: Andro specifically inhibited HIF-1α protein and gene accumulation under hypoxia.Notes: T47D and MDA-MB-231 cells were treated with or without Andro under hypoxia for 8 hours. The protein extracts from these cells were subjected to Western blotting for HIF-1α, HIF-2α, and HIF-1β expression. β-Actin was used as a loading control. Representative Western blotting images of T47D cells (A) and MDA-MB-231 cells (B) are shown. The quantitative data (mean ± SD) of T47D cells (C) and MDA-MB-231 (D) cells were calculated from at least three separate experiments, taking the ratio of HIF-1α, HIF-2α, and HIF-1β bands to the β-actin band in 1% O2 group. *P<0.05 and **P<0.01 compared with hypoxia group (1% O2). (E) Total RNAs were extracted and HIF-1α mRNA levels were analyzed by qPCR. *P<0.05 and **P<0.01 compared with hypoxia group (1% O2).Abbreviations: Andro, Andrographolide; HIF-1, hypoxia-inducible factor-1; SD, standard deviation; mRNA, messenger RNA; qPCR, real-time polymerase chain reaction.

Mentions: The ability of Andro to inhibit HIF-1 expression was tested by evaluating the HIF-1 expression levels in the breast cancer cells T47D and MDA-MB-231 under hypoxia. As shown in Figure 2A–D, hypoxia induced HIF-1α expression in both T47D and MDA-MB-231 cells, and incubation with Andro (8 hours) led to a dose-dependent decrease of HIF-1α, but not HIF-2α or HIF-1β. The HIF-1α protein level was reduced to 0.3-fold (T47D) and 0.2-fold (MDA-MB-231) in the 1% O2 group (P<0.01) after 1 μM Andro treatment. To elucidate whether this decrease in HIF-1α protein level was caused by the effect of Andro on mRNA expression under our experimental conditions, qPCR assay was employed. Indeed, the addition of Andro significantly reduced HIF-1α mRNA level in a dose-dependent manner under hypoxia (Figure 2E). Thus, Andro blocks the hypoxia-induced accumulation of HIF-1α at the transcriptional stage by reducing HIF-1α mRNA level.


Andrographolide inhibits hypoxia-inducible factor-1 through phosphatidylinositol 3-kinase/AKT pathway and suppresses breast cancer growth.

Li J, Zhang C, Jiang H, Cheng J - Onco Targets Ther (2015)

Andro specifically inhibited HIF-1α protein and gene accumulation under hypoxia.Notes: T47D and MDA-MB-231 cells were treated with or without Andro under hypoxia for 8 hours. The protein extracts from these cells were subjected to Western blotting for HIF-1α, HIF-2α, and HIF-1β expression. β-Actin was used as a loading control. Representative Western blotting images of T47D cells (A) and MDA-MB-231 cells (B) are shown. The quantitative data (mean ± SD) of T47D cells (C) and MDA-MB-231 (D) cells were calculated from at least three separate experiments, taking the ratio of HIF-1α, HIF-2α, and HIF-1β bands to the β-actin band in 1% O2 group. *P<0.05 and **P<0.01 compared with hypoxia group (1% O2). (E) Total RNAs were extracted and HIF-1α mRNA levels were analyzed by qPCR. *P<0.05 and **P<0.01 compared with hypoxia group (1% O2).Abbreviations: Andro, Andrographolide; HIF-1, hypoxia-inducible factor-1; SD, standard deviation; mRNA, messenger RNA; qPCR, real-time polymerase chain reaction.
© Copyright Policy
Related In: Results  -  Collection

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f2-ott-8-427: Andro specifically inhibited HIF-1α protein and gene accumulation under hypoxia.Notes: T47D and MDA-MB-231 cells were treated with or without Andro under hypoxia for 8 hours. The protein extracts from these cells were subjected to Western blotting for HIF-1α, HIF-2α, and HIF-1β expression. β-Actin was used as a loading control. Representative Western blotting images of T47D cells (A) and MDA-MB-231 cells (B) are shown. The quantitative data (mean ± SD) of T47D cells (C) and MDA-MB-231 (D) cells were calculated from at least three separate experiments, taking the ratio of HIF-1α, HIF-2α, and HIF-1β bands to the β-actin band in 1% O2 group. *P<0.05 and **P<0.01 compared with hypoxia group (1% O2). (E) Total RNAs were extracted and HIF-1α mRNA levels were analyzed by qPCR. *P<0.05 and **P<0.01 compared with hypoxia group (1% O2).Abbreviations: Andro, Andrographolide; HIF-1, hypoxia-inducible factor-1; SD, standard deviation; mRNA, messenger RNA; qPCR, real-time polymerase chain reaction.
Mentions: The ability of Andro to inhibit HIF-1 expression was tested by evaluating the HIF-1 expression levels in the breast cancer cells T47D and MDA-MB-231 under hypoxia. As shown in Figure 2A–D, hypoxia induced HIF-1α expression in both T47D and MDA-MB-231 cells, and incubation with Andro (8 hours) led to a dose-dependent decrease of HIF-1α, but not HIF-2α or HIF-1β. The HIF-1α protein level was reduced to 0.3-fold (T47D) and 0.2-fold (MDA-MB-231) in the 1% O2 group (P<0.01) after 1 μM Andro treatment. To elucidate whether this decrease in HIF-1α protein level was caused by the effect of Andro on mRNA expression under our experimental conditions, qPCR assay was employed. Indeed, the addition of Andro significantly reduced HIF-1α mRNA level in a dose-dependent manner under hypoxia (Figure 2E). Thus, Andro blocks the hypoxia-induced accumulation of HIF-1α at the transcriptional stage by reducing HIF-1α mRNA level.

Bottom Line: Further, Andro inhibited T47D and MDA-MB-231 cell proliferation and colony formation.In addition, it exhibited significant in vivo efficacy and antitumor potential against the MDA-MB-231 xenograft in nude mice.In conclusion, these results highlighted the potential effects of Andro, which inhibits HIF-1, and hence may be developed as an antitumor agent for breast cancer therapy in future.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery, Beijing Chao-Yang Hospital, Beijing, People's Republic of China.

ABSTRACT
Hypoxia-inducible factor-1 (HIF-1) is a master regulator of the transcriptional response to hypoxia. HIF-1α is one of the most compelling anticancer targets. Andrographolide (Andro) was newly identified to inhibit HIF-1 in T47D cells (a half maximal effective concentration [EC50] of 1.03×10(-7) mol/L), by a dual-luciferase reporter assay. It suppressed HIF-1α protein and gene accumulation, which was dependent on the inhibition of upstream phosphatidylinositol 3-kinase (PI3K)/AKT pathway. It also abrogated the expression of HIF-1 target vascular endothelial growth factor (VEGF) gene and protein. Further, Andro inhibited T47D and MDA-MB-231 cell proliferation and colony formation. In addition, it exhibited significant in vivo efficacy and antitumor potential against the MDA-MB-231 xenograft in nude mice. In conclusion, these results highlighted the potential effects of Andro, which inhibits HIF-1, and hence may be developed as an antitumor agent for breast cancer therapy in future.

No MeSH data available.


Related in: MedlinePlus