【作者】 巴一； 吴雄志； 谢广茹； 陈丹； 王伟；

【Author】 Ba Yi Wu Xiong-Zhi Xie Guang-Ru Chen Dan Wang Wei Department of Medical Oncology, Tianjin Medical University Cancer Institute and Hospital, Tianjin, 300060, China Department of Integrative Oncology, Tianjin Medical University Cancer Institute and Hospital, Tianjin, 300060, China Tianjin Infection diseases Hospital, Tianjin 300192, China Tianjin Medical University, Tianjin, 300060, China.

【机构】 天津市肿瘤医院内科； 天津市肿瘤医院中西医结合科； 天津市传染病医院； 天津医科大学；

【摘要】 目的:守宫为壁虎科动物无蹼壁虎(Gekko swinhonis G u enther)或其它几种壁虎的全体,是传统的成寒软坚药,中医长期以来用以治疗癌肿。作为细胞外基质的重要组成部分,多糖在细胞的分化与增殖中具有重要作用。我们从传统抗癌中药守宫里分离出硫酸多糖Gepsin,观察其对人肝癌B el-7402细胞生长与分化的作用。方法:人肝癌 BEL-7402细胞、人早幼粒白血病HL-60细胞与人肝L-02 细胞在含10％小牛血清的RPMI-1640培养基中培养,分别于加药后30分钟、1-6天收集细胞,台盼蓝染色,计数活细胞与死细胞。取对数生长期B el-7402细胞,加药4天后,光学显微镜下观察细胞形态,收集细胞,透射电子显微镜观察细胞超微结构改变,流式细胞仪分析细胞凋亡与细胞周期, 收集上清,分别用放免法与比色法检测AFP与ALB细胞,细胞用不含胰酶的EDTA消化至镜下观察培养瓶中无细胞贴壁,台盼蓝染色,计数活细胞。结果:Gepsin可显著抑制肝癌Bel-7402细胞的生长而不抑制人早幼粒白血病HL-60细胞与人肝L-02细胞的增殖。Gepsin可使肝癌Bel-7402细胞的存活率有轻度的下降,但不影响L—02细胞的存活率。 Gepsin不能诱导B el-7402细胞凋亡(空白对照组与Gepsin 处理组凋亡率:12．00％±0．85％,13．10％±2．40％,P=0． 63)。Gepsin可使G2／M期细胞增加(空白对照组与Gepsin处理组G2／M期细胞比例:13．55％±0．07％,16．70％±0．28％, P=0．03)。光学显微镜下Gepsin作用后的B el-7402细胞呈纺锤状,透射电子显微镜下细胞线粒体减少,异染色质增加, 常染色质减少,核异型性减轻。Gepsin作用后的Bel-7402细胞AFP分泌下降,ALB分泌上升。Gepsin使Bel-7402细胞 G2／M期细胞增加。结论:Gepsin可显著抑制肝癌Bel-7402 细胞的增殖并诱导其分化,且对正常肝细胞无细胞毒作用。 Gepsin是中药守宫抗肿瘤的重要物质基础。更多还原

【Abstract】 Objective: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide. As it is difficulty to give sufficient dose due to the poor liver function and low sensitivity for the anti-cancer agents, chemotherapy adds little to the overall survival of HCC patients. Gekko swinhonis G_enther has been used as the anti-cancer drug in traditional Chinese medicine for hundreds of years. However, its anticancer components have not been reported. We separated sul-fated polysaccharide from Gekko swinhonis G_enther. Methods: HCC cell line (Bel-7402) and liver cell line (L-02) were exposed to Gekko sulfated polysaccharide (Gepsin) (100g/ml and 10g/ml). Cell proliferation and livability were determined by trypan blue stain. Apoptosis and cell cycle were detected by flow cytometry. Morphologic and ultrastructural variations were determined by optic and electronic microscopy. The secretion of alpha-fetoprotein (AFP) and albumin (ALB) was detected by rayeroimmunology. Results: Gepsin didn’ t suppress the proliferation of L-02 cells but inhibited that of Bel-7402 cells strongly. The livability of Bel-7402 cells was suppressed by Gepsin lightly, whereas Gepsin didn’ t inhibit the livability of L-02 cells. Significant blockade in the G2/M phase occurred in Bel-7402 cells 4 days after their exposure to Gepsin. Apoptosis percentage of Bel-7402 cells showed no significant difference between Gepsin-treated cells and PBS-treated cells. Treatment with Gepsin, Bel-7402 cells showed ultrastructural features of differentiation. The AFP secretion decreased while the ALB secretion increased markedly on Gepsin-treated cells (Table 1). Conclusion: Gepsin suppressed the proliferation and induced differentiation of liver cancer cells, but the toxicity to normal liver cells was negligible.更多还原