Mucin—4在百里醌抑制肝癌生长中的作用.docVIP

Mucin—4在百里醌抑制肝癌生长中的作用 　　[摘要] 目的 探讨百里醌对体内外肝癌生长的影响，并对其机制进行初步探讨。 方法 不同浓度百里醌作用人肝癌细胞株SMMC-7721后，采用四甲基偶氮唑蓝（MTT）比色法检测其对细胞生长的抑制作用；Hoechst染色后荧光显微镜下观察细胞形态学变化；RT-PCR检测Mucin-4 mRNA在SMMC-7721细胞中的表达；Western blotting检测肝癌细胞中Mucin-4蛋白表达；建立裸鼠肝癌皮下移植模型，完全随机法分为对照组和低（10 μmol/L）、中（20 μmol/L）、高剂量（40 μmol/L）百里醌组，观察不同剂量百里醌对裸鼠肝癌皮下移植瘤生长的影响；免疫组织化学法检测肿瘤组织中Mucin-4的阳性表达。 结果 不同浓度（10、20、40 μmol/L）百里醌作用人肝癌SMMC-7721 24 h后，细胞存活率分别为（80.14±9.84）%、（71.68±6.51）%和（64.58±5.24）%；百里醌作用后，SMMC-7721细胞出现典型凋亡形态学改变；百里醌干预后SMMC-7721细胞中Mucin-4蛋白和mRNA的表达均明显下调，呈浓度依赖性；实验结束后，对照组和低、中、高剂量百里醌组肿瘤体积分别为（1056.3±236.3）、（672.3±178.8）、（529.4±165.7）、（473.1±141.5） mm2，各百里醌组肿瘤体积明显低于对照组（P0.05），低、中、高剂量百里醌组抑瘤率分别为36.5%、49.9%和30.3%；百里醌可明显降低肿瘤组织中Mucin-4的阳性表达。 结论 百里醌可显著抑制体内外肝癌生长，该作用可能通过抑制Mucin-4的表达而实现。 　　[关键词] 肝癌；百里醌；Mucin-4 　　[中图分类号] R735.7 [文献标识码] A [文章编号] 1674-4721（2013）11（a）-0010-04 　　The role of Mucin-4 in the anti-tumor effect of thymoquinone on liver cancer 　　TANG Zhen 　　Yueyang Hospital，the Medical College of Xiangya，the Central South University，Yueyang 414000，China 　　[Abstract] Objective To investigate the effect and the mechanism of thymoquinone in the growth inhibition of liver cancer.Methods The human liver cancer cell line SMMC-7721 was treated with different concentrations of thymoquinone，the cellular proliferation was assessed by methyl thiazolyl tetrazolium （MTT） method.SMMC-7721 cells morphological changes were observed under the fluorescence microscopy after Hoechst staining.The expression of Mucin-4 mRNA was detected by RT-PCR.Western blotting was used to detect the expression of Mucin-4 in liver cancer cells.Furthermore，human liver cancer model was set up by using SMMC-7721 cells.While the tumors grew with diameter of approximate 3 mm，mice were randomized into the control group，low dose thymoquinone group，middle dose thymoquinone group，high dose thymoquinone group.All treatment lasted for three weeks，thrice per week.Six weeks after implantation，tumor weight and inhibition rate was evaluated respectively after th