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目的:本文旨在超声提取法汉黄芩素并研究汉黄芩素在体外对人肝癌肿SMMC-7721细胞的增殖影响及其分子作用机制。方法 :超声提取法提纯汉黄芩素;应用CCK8法检测不同浓度汉黄芩素对K562、PC-3、MDA-MB-231和SMMC-7721细胞增殖的影响;DAPI荧光法检测SMMC-7721细胞凋亡;流式细胞仪检测细胞中钙离子浓度变化;流式细胞仪检测细胞膜电位变化;westernblot法检测细胞中ERK1/2蛋白的表达水平。结果 :(1)0~300μmol/L的汉黄芩素对K562、PC-3、MDA-MB-231和SMMC-7721肿瘤组织细胞具有显著的生长增殖抑制作用,且剂量越大时间越长抑制效果越好。(2)DAPI荧光显微镜检测结果表明,汉黄芩素作用后的SMMC-7721的细胞形状发生明显变化,细胞核破裂,染色质浓缩,可见明显的细胞碎片。结论 :汉黄芩素能够显著性抑制SMMC-7721肿瘤组织细胞的增殖,且分子作用机制为影响ERK1/2蛋白的表达水平的信号通路诱导凋亡。
OBJECTIVE: The purpose of this study is to study the effects of wogonin on the proliferation of human hepatoma SMMC-7721 cells and its molecular mechanism in vitro. Methods: Ultrasonic extraction was used to purify wogonin. The effects of different concentrations of wogonin on the proliferation of K562, PC-3, MDA-MB-231 and SMMC-7721 cells were detected by CCK8 assay. The apoptosis of SMMC-7721 cells was detected by DAPI The changes of Ca2 + concentration in cells were detected by flow cytometry. The changes of cell membrane potential were detected by flow cytometry. The expression of ERK1 / 2 protein was detected by western blot. Results: (1) Wogonin at 0 ~ 300μmol / L had significant inhibitory effects on the growth of K562, PC-3, MDA-MB-231 and SMMC-7721 tumor cells, and the longer the dose, the longer the inhibitory effect The better. (2) The results of DAPI fluorescence microscopy showed that the cell shape of SMMC-7721 after Wogonin treatment significantly changed, the nucleus ruptured, and the chromatin condensed, showing obvious cell debris. Conclusion: Wogonin can significantly inhibit the proliferation of SMMC-7721 tumor cells, and the molecular mechanism of apoptosis is signal pathway that induces the expression of ERK1 / 2 protein.