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守宫硫酸多糖对肝癌SMMC-7721细胞分化和增殖的影响

辛亮1,吴雄志1,谢广茹1,2   

  1. 1. 天津医科大学附属肿瘤医院
    2.
  • 收稿日期:2011-03-07 修回日期:2011-06-21 出版日期:2011-12-15 发布日期:2011-12-15
  • 通讯作者: 辛亮

Effects of Gekko sulfated polysaccharide on the proliferation and differentiation of hepatic cancer cell line SMMC-7721

  • Received:2011-03-07 Revised:2011-06-21 Published:2011-12-15 Online:2011-12-15

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摘要： 目的：研究守宫硫酸多糖（Gepsin）对肝癌SMMC-7721细胞分化和增殖的影响，并进一步探讨其可能的机制。方法：将不同浓度的Gepsin加入对数生长的细胞SMMC-7721进行培养，在不同的时间利用台盼蓝染色观察细胞的增殖情况。同时，收集细胞培养上清液，利用联合放免，以溴钾酚绿法分别观察Gepsin对甲胎蛋白（AFP）、白蛋白（ALB）分泌的影响；利用酶联免疫吸附试验法观察Gepsin对转化生长因子（TGF）-β1、血管内皮生长因子（VEGF）的分泌的影响；利用光镜观察Gepsin对SMMC-7721细胞形态的影响。结果：随着处理组Gepsin的浓度升高，SMMC-7721细胞的增殖明显受到抑制；而对细胞活率无明显影响。Gepsin可增加培养上清中的ALB分泌量，降低AFP分泌量。光学显微镜下可见加入Gepsin后细胞的形态由圆形变为纺锤形。Gepsin对培养上清分泌的VEGF无影响，但可使TGF-β1增加。结论：Gepsin可明显抑制肝癌细胞SMMC-7721的增殖，诱导 SMMC-7721细胞分化。其机制可能通过上调TGF-β1来诱导肝癌细胞分化实现的。

关键词: 守宫, 多糖类, 细胞系, 肿瘤, 肝肿瘤, 癌, 甲胎蛋白类, 白蛋白类

Abstract: Abstract Objective: To observe Gepsin on the proliferation and differentiation of hepatic cancer cell line SMMC-7721 .Methods: SMMC-7721 cells were cultured in RPM-1640 medium supplemented with 10% fetal bovine serum at 37℃ in a humidified 5% CO2 incubator. SMMC-7721 cells were seeded in 24-well plates at 1×104 cells/ml .Cells were allowed to grow for one day before being exposed to Gepsin(100μg/ml and 10μg/ml, respectively). SMMC-7721 cells were collected at 30min,then 24, 48, 72, 96, 120, 144h after treatment with Gepsin. Trypan blue stain were used to determine total cell count and viable cell number. SMMC-7721 cells were seeded in 24-well plates at 1×104 cells/ml .Cells were allowed to grow for one day before being exposed to Gepsin(100μg/ml and 10μg/ml, respectively)，At day 7 after treatment with drugs, culture medium was harvested. Then AFP and ALB were detected by rayeroimmunology. At day 7after treatment with drugs, SMMC-7721 cells were examined by light microscopy. The level of VEGF、TGF-β1 supernatant was detected by ELISA. Results: Hepatocarcinoma cell line (SMMC-7721 )were exposed to Gepsin(100μg/ml and 10μg/ml). Gepsin strongly inhibited the proliferation of hepatocarcinoma SMMC-7721 cells. While Gepsin did not have effect on viability of SMMC-7721 cells. Treated with Gepsin, SMMC-7721 cells showed ultrastructural features of differentiation. AFP secretion decreased while ALB secretion increased markedly on Gepsin-treated cells. SMMC-7721 cells showed a round shape under light microscope, while cells changed to spindle shape after exposure to Gepsin. The level of TGF-β1 significantly increased in Gepsin group (100μg/ml) （P<0.01）, the level of VEGF in suramin treated groups significantly increased than control group. Conclusion:⑴Gepsin strongly inhibited the proliferation of hepatocarcinoma SMMC-7721 cells. ⑵ The viability of SMMC-7721 cells was not suppressed by Gepsin. ⑶Gepsin could induce differentiation of SMMC-7721 cells.⑷Gepsin maybe induce the differentiation of SMMC-7721 cells via TGF-β1 ways.

Key words: 白蛋白类