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目的:结合体内外实验研究南蛇藤提取物对人肝癌细胞Hep G2上皮间质转化的作用及分子机制。方法:取对数生长期Hep G2细胞,除对照组外,分别给予南蛇藤提取物10、20、40、80、160μg·m L-1培养,采用Western Blotting法检测上皮间质转化(EMT)相关蛋白的表达水平;斑马鱼胚胎,于受精后48 h开始给药,正常组给予E3缓冲液,DMSO组为含1%DMSO的E3缓冲液,南蛇藤组各药物浓度分别为10、20、40、80、160μg·m L-1;给药24 h后,以Western Blotting法检测m TOR信号通路相关蛋白的表达水平。结果:与对照组相比,南蛇藤提取物各浓度组明显增加E-cadherin的表达量,降低vimentin和m TOR信号通路相关蛋白的表达水平。结论:南蛇藤提取物能在一定程度上逆转人肝癌细胞EMT,其分子机制可能与m TOR信号通路相关,m TOR可作为临床治疗肝癌的新靶点。
OBJECTIVE: To study the effect and molecular mechanism of Celastrus orbiculatus extract on Hep G2 epithelial-mesenchymal transition in vitro and in vivo. Methods: The Hep G2 cells in logarithmic growth phase were cultured except 10, 40, 40, 160 and 160μg · m L-1 of Celastrus orbiculatus respectively. Western Blotting was used to detect the expression of epithelial-mesenchymal transition (EMT ) Zebrafish embryos were injected 48h after fertilization, E3 buffer was given to the normal group, E3 buffer contained 1% DMSO in the DMSO group, and the drug concentration in the Celastrus group was 10, 20, 40, 80 and 160μg · m L-1 respectively. After 24 h, the expression of mTOR signaling pathway related protein was detected by Western Blotting. Results: Compared with the control group, the concentration of E-cadherin in each concentration of Celastrus ornithocarpa extract significantly decreased the expression of vimentin and mTOR signaling pathway related proteins. CONCLUSION: The extract of Celastrus orbiculatus can reverse the EMT of human hepatocellular carcinoma cells to a certain extent. Its molecular mechanism may be related to the mTOR signaling pathway. M TOR can be used as a new target for clinical treatment of hepatocellular carcinoma.