Polysaccharide of Boschniakia rossica induces apoptosis on laryngeal carcinoma Hep2 cells |
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Authors: | Zhenghui Wang Chuangxin Lu Caiqin Wu Min Xu Xiaohui Kou Demin Kong Gangli Jing |
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Institution: | 1. Department of Otolaryngology-Head and Neck Surgery, The Second Hospital, Xi''an Jiaotong University, Xi''an 710004, China;2. Cancer Center of Henan Province People''s Hospital, Zhengzhou 450000, China;3. Department of Surgery and Anesthesia, The Second Hospital, Xi''an Jiaotong University, Xi''an 710004, China |
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Abstract: | The aim of this study was to explore the anti-tumor potential of a polysaccharide isolated from Boschniakia rossica (BRP) in Hep2 human larynx squamous carcinoma cells. High performance size-exclusion chromatography analysis showed that BRP was a homogeneous polysaccharide and had a molecular weight of 22 kDa. Total carbohydrate content in BRP was determined to be 96.9%, without the presence of protein and nucleic acid. BRP suppressed the proliferation of Hep2 cells in a time- and dose-dependent manner. Cell cycle analysis revealed that exposure to BRP (200 μg/ml) caused a G0/G1 cell cycle arrest in Hep2 cells. Moreover, treatment with BRP at 100–400 μg/ml for 24 h induced a significant apoptosis Hep2 cells compared to untreated control cells, as determined by flow cytometry with annexin-V/propidium iodide double staining. Additionally, BRP treatment promoted the cleavage of pro-caspase-3, pro-caspase-8, and pro-caspase-9, coupled with increased expression of death receptor DR5 and Bax and reduced expression of Bcl-2. Taken together, our data demonstrate that BRP shows potent anti-tumor activity in human larynx squamous carcinoma, largely through induction of G0/G1 cell cycle arrest and apoptosis. Activation of both mitochondria-mediated and death receptor-mediated apoptosis pathways is involved in the cytotoxicity of BRP. |
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Keywords: | BRP boschniakia rossica SCC squamous cell carcinoma TCMs traditional Chinese medicines B rossica boschniakia rossica OVA ovalbumin MTT 3-(4 5-dimethylthiazol-2-yl)-2 5-diphenyltetrazolium bromide ConA concanavalin A LPS lipopolysaccharide BSA bovine serum albumin DMSO dimethyl sulfoxide FCS fetal calf serum DEAE diethylaminoethyl FITC fluorescein isothiocyanate TBS Tris buffered solution HPGPC high performance size-exclusion chromatography |
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