Andrographolide inhibits breast cancer through suppressing COX-2 expression and angiogenesis via inactivation of p300 signaling and VEGF pathway

Abstract

BackgroundAndrographolide (Andro), a diterpenoid lactone, has been used for treatment of various cancers with less adverse effects. However, the underlying mechanisms regarding its anti-tumor mechanism still remain unclear.MethodsCell viability and proliferation were measured by CCK8 and CFSE dilution assay. The localization of p50/p65 or cytochrome c was determined using confocal immunofluorescence. Streptavidin-agarose pulldown or ChIP assays were used to detect the binding of multiple transactivators to COX-2 promoter. The promoter activity was examined by a dual-Luciferase reporter assay. The functions of Andro on COX-2-mediated angiogenesis were also investigated using human HUVEC cells through tube formation and spheroids sprouting assay. The in vivo anti-tumor efficacy of Andro was analyzed in xenografts nude mice.ResultsThe results indicated that Andro could significantly inhibit the proliferation of human breast cancers, and suppress COX-2 expression at both protein and mRNA levels. Furthermore, Andro could dose-dependently inhibit COX-2-mediated angiogenesis in human endothelial cells. We have also found that Andro significantly promoted the activation of cytochrome c and activated caspase-dependent apoptotic signaling pathway. Our further explorations demonstrated that Andro inhibited the binding of the transactivators CREB2, C-Fos and NF-κB and blocked the recruitment of coactivator p300 to COX-2 promoter. Moreover, Andro could effectively inhibit the activity of p300 histone acetyltransferase (HAT), thereby attenuating the p300-mediated acetylation of NF-κB. Besides, Andro could also dramatically inhibit the migration, invasion and tubulogenesis of HUVECs in vitro. In addition, Andro also exhibited effective anti-tumor efficacy as well as angiogenesis inhibition in vivo.ConclusionIn current study, we explore the potential effects of Andro in suppressing breast cancer growth and tumor angiogenesis, as well as the precise mechanisms. This work demonstrated the potential anti-cancer effects of Andro, indicating that Andro could inhibit COX-2 expression through attenuating p300 HAT activity and suppress angiogenesis via VEGF pathway, and thereby could be developed as an antitumor agent for the treatment of breast cancer.

Andrographolide induces apoptosis in human osteosarcoma cells via the ROS/JNK pathway

“…Thus, novel treatments are required to improve the outcomes of patients with osteosarcoma. Andrographolide has been reported to be a promising therapeutic for treatment of multiple types of cancer, with potent antitumor effects and minimal toxicity (11,29,(39)(40)(41)(42). However, little is known about the effects and biochemical mechanisms of action of andrographolide in osteosarcoma.…”

Bruceine D inhibits tumor growth and stem cell‐like traits of osteosarcoma through inhibition of STAT3 signaling pathway

“…Carboxyfluorescein Succinimidyl Ester (CFSE) dilution assay was employed to detect cell proliferation as described previously . Osteosarcoma cells were labeled by CFSE using the CellTrace CFSE cell proliferation kit obtained from Thermo Fisher Scientific following the manufacturer's instructions and seeded into six‐well plates.…”

“…Carboxyfluorescein Succinimidyl Ester (CFSE) dilution assay was employed to detect cell proliferation as described previously. 21 Osteosarcoma cells were labeled by CFSE using the CellTrace CFSE cell proliferation kit obtained from Thermo Fisher Scientific following the manufacturer's instructions and seeded into six-well plates. After being cultured overnight, the labeled cells were stimulated with BD for 24, 48, or 72 hours, and cell proliferation index was detected and analyzed by flow cytometry (BD Biosciences).…”

CDK4 as a phytochemical based anticancer drug target

“…), mangiferin (Núñez Selles, Daglia, and Rastrelli 2016), voacangine (Y. Kim, Jung, and Kwon 2012), quinic acid (Singh, Chauhan, and Tripathi 2018), andrographolide (Peng et al 2018), luteolin (Cook 2018), apigenin (Yan et al 2017), rutin (Khan et al 2019), and gallic acid (Liu et al 2012) are already reported for their anticancer activities. This suggests that the screening process from the traditionally used medicinal plants contain both potential anticancer properties.…”