毛蕊花糖苷通过下调TGF-β抑制胶质母细胞瘤上皮间质转化的实验研究

来源 :中华神经医学杂志 | 被引量 : 0次 | 上传用户:a5477011266
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目的:探讨毛蕊花糖苷(VB)对胶质母细胞瘤细胞上皮间质转化的作用及分子机制。方法:(1)实验1:常规培养胶质母细胞瘤T98和U251细胞并各自分为4组,依次添加0、20、40、80 μmol/L的VB处理24 h。采用CCK-8实验检测4组细胞存活率;采用Transwell实验检测4组细胞迁移、侵袭情况;采用RT-PCR实验及Western blotting实验检测4组细胞转化生长因子-β(TGF-β)、波形蛋白、Snail蛋白mRNA及蛋白表达情况。(2)实验2:T98、U251细胞各分为4组,依次为对照组、VB组、TGF-β组和TGF-β+VB组。对照组及VB组细胞转染空载质粒、TGF-β组及TGF-β+VB组细胞转染TGF-β质粒;转染24 h后VB组、TGF-β+VB组细胞均加入40 μmol/L VB继续培养24 h,随后进行后续实验(迁移和侵袭实验、Western blotting实验,实验方法及检测指标同实验1)。(3)体内实验:裸鼠皮下种植U251细胞后分为2组,每组8只。实验组每天腹腔注射100 mg/kg的VB,对照组注射等量的PBS。记录小鼠肿瘤体积大小和小鼠体质量的变化;实验终点(移植后21 d)时,检测小鼠肿瘤重量并采用Western blotting实验检测肿瘤中TGF-β、波形蛋白、Snail的表达。结果:(1)实验1:与0 μmol/L VB组相比较,20 μmol/L、40 μmol/L VB组的细胞存活率差异无统计学意义(n P>0.05),而80 μmol/L VB组细胞存活率下降,差异有统计学意义(n P0.05), while that in the 80 μmol/L VB group showed significant difference (n P<0.05). As compared with that in the 0 μmol/L VB group, the cell number migrating and invading into the lower chamber in the 20 and 40 μmol/L VB groups was significantly smaller as concentration increased, and the mRNA and protein expressions of TGF-β, Snail and vimentin were also downregulated successively, with signficant differences (n P<0.05). (2) As compared with the control group, the cell number that migrating and invading into the lower chamber, and protein expressions of TGF-β, vimentin, and Snail in the VB group were significantly decreased (n P<0.05); while as compared with the control group, the cell number that migrating and invading into the lower chamber, and protein expressions of TGF-β, vimentin, and Snail in the TGF-β group were significantly increased (n P<0.05); as compared with those in the VB group, the cell number that migrating and invading into the lower chamber, and protein expressions of TGF-β, vimentin, and Snail in the TGF-β and TGF-β+VB groups were significantly increased (n P<0.05). (3) As compared with the control group, the tumor volume and tumor weight in the VB group were statistically decreased, and the TGF-β, vimentin and Snail protein expressions in the VB group were significantly downregulated (n P0.05).n Conclusion:VB inhibits epithelial-mesenchymal transformation in glioblastoma by suppressing the TGF-β expression.
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