Involvement of the mitochondrial pathway in bruceine D-induced apoptosis in Capan-2 human pancreatic adenocarcinoma cells
School of Chinese Medicine, Faculty of Science, The Chinese University of Hong Kong, Shatin, NT, Hong Kong SAR, P.R. China.International Journal of Molecular Medicine (Impact Factor: 2.09). 04/2012; 30(1):93-9. DOI: 10.3892/ijmm.2012.980
The fruit of Brucea javanica L. is a common herb used in Chinese medicine for the treatment of a variety of cancers. Our research group has previously identified bruceine D (BD), a quassinoid found abundantly in B. javanica, to have potent cytotoxic effect on a number of pancreatic cancer cell lines, including Panc-1, SW1990 and Capan-1 cells. In the present study, we showed that BD was also able to inhibit the growth of the Capan-2 human pancreatic adenocarcinoma cell line, but it exerted only modest cytotoxicity on the WRL68 human hepatocyte cell line and a human pancreatic progenitor cell line. The antiproliferative effects of BD were comparable to those exhibited by camptothecin and gemcitabine in our culture system. We found a dose-dependent decrease of the mitochondrial membrane potential in BD-treated Capan-2 cells as measured by the JC-1 assay. BD exposure was able to attenuate the expression of Bcl-2 protein in Capan-2 cells as detected by western blot analysis. In addition, the expression of both caspase 9 and caspase 3 in BD-treated Capan-2 cells was significantly accentuated. Moreover, BD was capable of inducing the fragmentation of genomic DNA in Capan-2 cells as evidenced by Hoechst staining. Cell cycle analysis demonstrated that BD could increase the percentage of Capan-2 cells in the subG1 phase in a dose-related manner. An increase in the apoptosis of Capan-2 cells was also observed by Annexin V and PI staining. These results unequivocally indicate that BD induces cytotoxicity in Capan-2 cells via the induction of cellular apoptosis involving the mitochondrial pathway.
Conference Paper: Apoptosis Activity of Brucea javanica Extract in Oral Cancer[Show abstract] [Hide abstract]
ABSTRACT: Objectives: The aim of this research is to identify apoptosis activity of Brucea javanica extract on oral cancer cell lines (HSC-2 and HSC-3) through cleaved caspase-3 and -9 Methods: Brucea javanica seed was dried at room temperature and extracted with ethanol for 24 hour. The extract were divided into different concentration (500 μg/ml, 100 μg/ml and 10 μg/ml) by serial dilution. Oral cancer cell lines (HSC-2 and HSC-3) were routine maintain in DMEM high glucose medium supplemented with 10% FBS and 1% penicillin-streptomycin in 5% CO2 and 37 C temperature. Six different treatment on cell lines were done with untreated, treated by DMSO as a negative control, Doxorubicin as a positive control, and Brucea javanica in different concentration respectively. On the next day cells were lysed and the express of cleaved Caspase-3 and 9 were analyzed with Western Blot. Results: Untreated and treated HSC-2 and HSC-3 cell lines with DMSO were grown well. The oral cancer cell lines that were treated by Doxorubicin 10 mM; Brucea javanica 500 g/ml; Brucea javanica 100 g/ml ; Brucea javanica 10 g/ml were floating and losing the attachment of the disk. Brucea javanica extract exhibited a dose dependent augmentation of apoptosis in all three cell lines. Western blot or Immunoblot is used to elucidate the specific anticancer mechanisms of the Brucea javanica extract, cleaved Caspase-3 and cleaved Caspase-9 were expressed/identified which shows the increasing of apoptosis activity. Conclusions: Brucea javanica induces apoptosis on HSC-2 and HSC-3 cell lines through cleaved caspase-3 and -925th IADR-SEA Division Annual Scientific Meeting 2011; 10/2011
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ABSTRACT: Brucea javanica (L.) Merr. is a medicine plant distributed widely throughout Asia where its bitter fruits have been used traditionally in medicine for treating various ailments and controlling some pests. In recent years, concerns over the potential impact of synthetic pesticides on human health and environment have now become more pressing to develop environmentally friendly pesticides. In this paper, brusatol, a quassinoid, was isolated from the fruit of B. javanica, and identified using X-ray crystallographic analysis. Results showed that brusatol has potent contact toxicity (LD50, 2.91 μg/larva, 72 h) and anfieedant activity (AFC50, 17.4 mg/L, 48 h) against the third-instar larvae of Spodoptera exigua. Brusatol demonstrated cytotoxic effects to the tested insect cell lines, IOZCAS-Spex-II and Sf21, in a time- and dose-dependent manner. After brusatol treatment, apoptotic cell death with the DNA fragmentation, activation of caspase-3 and release of cytochrome c was preliminarily observed in both IOZCAS-Spex-II and Sf21. These results indicated the existence of apoptotic death with the mitochondrial-dependent pathway induced by brusatol in Sf21 and IOZCAS-Spex-II cell lines. Our studies will provide important knowledge to understand mechanisms of action of brusatol and to develop brusatol and its derivatives as insecticides.Pesticide Biochemistry and Physiology 09/2013; 107(1):18–24. DOI:10.1016/j.pestbp.2013.04.007 · 2.01 Impact Factor
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ABSTRACT: Pancreatic cancer has a poor prognosis with a 5-year survival rate of <5%. It does not respond well to either chemotherapy or radiotherapy, due partly to cancer cell apoptotic resistance (AR). AR has been attributed to certain genetic abnormalities or defects in apoptotic signaling pathways. In pancreatic cancer, significant mutations of K-ras and p53, constitutive activation of NFκB, over-expression of heat shock proteins (Hsp90, Hsp70), histone deacetylase (HDACs) and the activities of other proteins (COX-2, Nrf2 and bcl-2 family members) are closely linked with resistance to apoptosis and invasion. AR has also been associated with aberrant signaling of MAPK, PI3K-AKT, JAK/STAT, SHH, Notch, and Wnt/β-catenin pathways. Strategies targeting these signaling molecules and pathways provide an alternative for overcoming pancreatic cancer AR. The use of herbal medicines or natural products (HM/NPs) alone or in combination with conventional anti-cancer agents has been shown to produce beneficial effects through actions upon multiple molecular pathways involved in AR. The current standard first-line chemotherapeutic agents for pancreatic cancer are gemcitabine (Gem) or Gem-containing combinations; however, the efficacy is dissatisfied and this limitation is largely attributed to resistance to apoptosis. Meanwhile, emerging data have pointed to a combination of HM/NPs that may augment the sensitivity of pancreatic cancer cells to Gem. Greater understanding of how these compounds affect the molecular mechanisms of apoptosis may propel development of HM/NPs as anti-cancer agents and/or adjuvant therapies forward. In this review, we give a critical appraisal of the use of HM/NPs alone and in combination with anti-cancer drugs. We also discuss the potential regulatory mechanisms whereby AR is involved in these protective pathways.The International Journal of Biochemistry & Cell Biology 05/2014; 53. DOI:10.1016/j.biocel.2014.05.021 · 4.05 Impact Factor
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