Phenolic alkaloids from Menispermum dauricum inhibits BxPC-3 pancreatic cancer cells by blocking of Hedgehog signaling pathway

Abstract

Background:The Hedgehog (Hh) signaling pathway plays an important role in pancreatic cancer (PC) cells. Phenolic alkaloids from Menispermum dauricum (PAMD), a traditional Chinese medicine used for the treatment of immune disorders, have been reported to have antitumor activity recently.Objective:To investigate the efficacy and mechanism of PAMD against PC cell BxPC-3.Materials and Methods:F assay was used to assess cell proliferation inhibition of PAMD; the apoptotic induction and cell cycle arrest was detected by flow cytometry; the BxPC-3 xenograft was established to evaluate the tumor growth inhibition of PAMD; hematoxylin-eosin staining was applied to analyze the pathological morphology of tumor tissues; immunohistochemistry (IHC) and Western blot was adopted to detect the protein levels; quantitative real-time polymerase chain reaction was used to determine the mRNA expressions.Results:PAMD shows time-and dose-dependent proliferation inhibition on the BxPC-3 cell, induced G0/G1 phase arrest and cell apoptosis in vitro. PAMD also showed better inhibition of tumor growth and a preferable safety profile compared with chemotherapeutic regimen 5-fluoro-2, 4 (1 H, 3 H) pyrimidinedione in BxPC-3 xenograft in vivo. Furthermore, PAMD directly decreases the protein and mRNA levels of Sonic Hedgehog (Shh) and its downstream transcription factor Gli-1 in the BxPC-3 tumor tissues.Conclusion:The treatment of PAMD displayed Hh signaling pathway blockade through decreasing the protein and mRNA levels of Shh and its downstream transcription factor Gli-1, suggesting a promising strategy in treating human PC.

Dauricine suppresses the growth of pancreatic cancer in�vivo by modulating the Hedgehog signaling pathway

“…The tumor volume was calculated using the following formula: Volume=(4π/3) × (L/2) 3 , where L is the mean tumor length measured in 3 dimensions. It was observed that maximum tumor diameter was ~1.2 cm, and the tumor volume in the saline control group reached ~1,000 mm 3 at 21 days, which was in accordance with the results from our previous study (30). The weights of the tumor and spleen were measured.…”

“…Dauricine, a bioactive component extracted from the moonseed rhizome, has demonstrated promising pharmacological attributes in clinical settings (24,30). It has long been used as a traditional Chinese medicine remedy for inflammatory disorders.…”

“…A previous study on colon cancer in mice used 40 mg/kg dauricine injected every 2 days until day 9 (24), which showed that dauricine significantly suppressed colonic tumor growth. Our previous study demonstrated the significant in vivo effects of a 21-day treatment with 10 and 20 mg/kg phenolic alkaloid mixture on pancreatic cancer (30). Therefore, a 21-day treatment with two doses of dauricine (6 and 12 mg/kg body weight) was tested in the present study.…”

“…The pancreatic cancer BxPC-3 ×enograft animal model was established according to previously described methods (30). Cell suspension (0.2 ml; 5×10 6 cells/ml) was inoculated subcutaneously into the right armpit of the nude mice.…”

“…Dauricine also attenuates inflammatory reactions (25), exerts neuroprotective effects (26,27), and blocks potassium and calcium channels (28,29). A previous study by our group examined the effects of a mixture of phenolic alkaloids containing daurisolin, dauricine, daurinoline and dauricicoline in pancreatic cancer BxPC-3 cells and BxPC-3 ×enograft mice (30). The mixture exhibited a time-and dose-dependent inhibitory effect on the proliferation of the BxPC-3 cells, and induced G 0 /G 1 phase arrest and cell apoptosis; it also inhibited tumor growth in the BxPC-3 ×enograft in vivo .…”