Chen Qun, Yang Gui-Wen, An Li-Guo
Department of Biology, Shandong Normal University, Jinan 250014, Shandong Province, China.
World J Gastroenterol. 2002 Oct;8(5):832-6. doi: 10.3748/wjg.v8.i5.832.
To study the apoptosis of hepatoma cells SMMC-7721 induced by polysaccharide isolated from Ginkgo biloba seed.
Ginkgo biloba seed polysaccharide (GBSP) was isolated by ethanol fractionation of Ginkgo biloba seed and purified by Sephadex G-200 chromatography. The purity of GBSP was verified by reaction with iodine-potassium iodide and ninhydrin and confirmed by UV spectrophotometer, cellulose acetate membrane electrophoresis and Sepharose 4B gel filtration chromatography. The Scanning Electron Microscope (SEM) and Flow Cytometry (FCM) were used to examine the SMMC-7721 cells with and without GBSP treatment at 500 mg/ml for 36 h.
GBSP product obtained was of high purity with the average molecular weight of 1.86 X 10(5). Quantitative analysis of SMMC-7721 cells in vitro with FCM showed that the percentages of G(2)-M cells without and with GBSP treatment were 17.01+/-1.28 % and 11.77+/-1.50% (P<0.05), the debris ratio of the cells were 0.46+/-0.12 % and 0.06+/-0.06 % (P<0.01), and the apoptosis ratio of cells was 3.84+/-0.55 % and 9.13+/-1.48 % (P<0.01) respectively. Following GBSP treatment, microvilli of SMMC-7721 cells appeared thinner and the number of spherical cells increased markedly. Most significantly, the apoptosis bodies were formed on and around the spherical cells treated with GBSP.
GBSP could potentially induce the apoptosis of SMMC-7721 cells.
研究银杏种仁多糖诱导肝癌细胞SMMC - 7721凋亡的作用。
采用乙醇分级沉淀法从银杏种仁中分离银杏种仁多糖（GBSP），并经Sephadex G - 200柱色谱法纯化。通过与碘 - 碘化钾及茚三酮反应、紫外分光光度计、醋酸纤维素膜电泳和Sepharose 4B凝胶过滤色谱法验证GBSP的纯度。运用扫描电子显微镜（SEM）和流式细胞术（FCM）检测500 mg/ml GBSP处理36 h的SMMC - 7721细胞及未处理的细胞。
获得的GBSP产品纯度高，平均分子量为1.86×10⁵。FCM对体外培养的SMMC - 7721细胞进行定量分析显示，未处理组和GBSP处理组G₂ - M期细胞百分比分别为17.01±1.28%和11.77±1.50%（P<0.05），细胞碎片率分别为0.46±0.12%和0.06±0.06%（P<0.01），细胞凋亡率分别为3.84±0.55%和9.13±1.48%（P<0.01）。GBSP处理后，SMMC - 7721细胞的微绒毛变细，球形细胞数量显著增加。最显著的是，GBSP处理的球形细胞上及周围形成了凋亡小体。
GBSP可能诱导SMMC - 7721细胞凋亡。
