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Briefly, Acinar cell carcinoma (ACC) tumor cells (1 × 105 cells/mL) were grown in Roswell Park Memorial Institute (EPMI) 1640 medium supplemented with 10% fetal bovine serum (FBS) under atmosphere of 5% carbon dioxide at 37☌ for 72 h. MTT method was used for in vitro antitumor test. The WAF sample and the sonicated fractions were finally dried with freeze-dried. To obtain samples with different molecular weights, WAF was dissolved in pure water and ultrasonicated at room temperature with an ultrasonic cleaner (MUS/1004) for different time such as 0 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 16 h corresponding to the samples coded as WAF, WAF1, WAF2, WAF3, WAF4, WAF5, WAF6, and WAF7. WAF was isolated from fruit bodies of Auricularia auricula-judae according to our previously reported method. Male BALB/c mice were got from The Animal Centre, Tongji Medical College Huazhong University of Science and Technology (Wuhan, China). Sarcoma 180 (S-180) was supplied by Department of Pharmacy, Tongji Medical College Huazhong University of Science and Technology (Wuhan, China). Acinar cell carcinoma was provided by School of Stomatology, Wuhan University (Wuhan, China). ExperimentalĪuricularia auricula-judae was obtained from Fangxian (Hubei Province, China). The mechanism of the antitumor activities of the wAF was investigated by the morphological approaches (fluorescence microscopy with Hoechst stain) and immunohistochemical method, attempting to elucidate the antitumor mechanism. In the present work, in vivo antitumor activities of the polysaccharide wAF against xenograft Sarcoma 180 tumor cells and in vitro inhibition ratio to the proliferation of Acinar cell carcinoma(ACC) tumor cells were evaluated. However, its antitumor bioactivities have never been reported. It’s the first time to reveal conformation and structure of the most abundant polysaccharide contained in Auricularia auriculajudae. įigure 1: Molecular structure of polysaccharide wAF. Its chemical structure was determined to be composed of a backbone of α-(1→3)-linked D-mannopyranose residues with pendant side groups of β-D-xylose, β-D-glucose or β-D-glucuronic acid at position O6 or O2 ( Figure 1). In our previous work, the structure of a water soluble acidic heteropolysaccharide isolated from Auricularia auricula-judae was studied. Various polysaccharides isolated from Pleurotus tuber regium, Lentinus Edodes, Poria cocos, Ganoderma tsugae mycelium, Auricularia auricula-judae have been studied, focusing on the effects of the structure and chain properties such as flexibility, or multiplex formations on the bioactivities.
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These lectins have potential functions in immune surveillance by recognizing high mannose structures present on pathogenic organisms such as bacteria and fungi. Some mannose-binding proteins are C-type lectins, found in animal cell surface.
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In addition to the acidic character of the polysaccharide, the mannose rich structure might have beneficial effects as biomedical agents and natural drugs. Īcidic polysaccharide composed of α-1→3-D-mannopyranose as backbone is known to constitute the major part of water soluble fraction of Auricularia auricula-judae, an ear- shaped edible fungus. Many results show that polysaccharides have a broad spectrum of biological effects, such as antibiotic, anti-mutant, anticoagulant and immunostimulant activities. Polysaccharides have attracted more and more attention recently because of their antitumor and immunomodulation effects. These findings confirmed that the wAF polysaccharide had good antitumour activity and could be considered as a potential antitumor agent KeywordsĪuricularia auricula-Judae, Polysaccharide, Antitumor activity, Apoptosis IntroductionĬancer is a leading cause of death worldwide resulting from a mutation in the chromosomal DNA of a normal cell. Immunohistochemistry for apoptosis-related proteins Bax and Bcl-2 expression revealed that wAF3 induced S-180 tumour cell apoptosis by up-regulation of Bax and down-regulation of Bcl- 2. Fluorescence microscopy and tumour tissues cell morphology observation indicated that wAF3 fraction induced apoptosis of tumour cell. Moreover, the enhancement ratios of mice body weight for all wAF fractions were higher than that for 5-Fluorouracil, which is a chemotherapeutic anticancer agent. It significantly inhibited tumour growth in dose-dependent way in BALB/c mice. WAF fractions exhibited strong inhibition against Acinar cell carcinoma proliferation in vitro test. It composed of a backbone of α- (1→3)-linked D-mannopyranose residues. A water soluble acidic heteropolysaccharide named wAF was isolated from Auricularia auricula-judae.