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目的:初步探讨岩黄连总碱对人肺腺癌A549细胞增殖抑制及诱导凋亡的作用,及其可能的作用机制。方法:将肺癌A549细胞株在体外条件下加入不同质量浓度岩黄连总碱。用四甲基偶氮唑蓝比色(MTT)法检测不同浓度(0.1,0.05,0.01,0.005 g·L-1)岩黄连总碱分别作用24,48,72 h后,对人肺腺癌A549细胞的生长抑制情况。采用Hoechst染色法检测岩黄连总碱处理A549细胞48 h的生长情况。流式细胞术检测岩黄连总碱不同药物浓度诱导凋亡的情况。RT-PCR法检测岩黄连及顺铂对A549细胞中Caspase-3,Survivin mRNA的表达影响变化情况。结果:MTT结果提示岩黄连总碱对肺癌A549细胞增殖具有一定的抑制作用,且抑制作用呈现时间-剂量关系。MTT及Hoechst染色显示岩黄连总碱组较阴性组明显抑制A549细胞增殖(P<0.05)。流式细胞仪检测结果显示岩黄连诱导A549细胞凋亡作用随着药物浓度的增加,凋亡率升高。RT-PCR结果提示岩黄连组较阴性组上调了Caspase-3 mRNA的表达,下调了Survivin mRNA的表达。结论:岩黄连总碱具有一定的抑制人肺腺癌A549细胞增殖和诱导凋亡作用,并呈时间-剂量关系。其机制可能与上调Caspase-3的表达和下调Survivin的表达有关。
OBJECTIVE: To investigate the inhibitory effect of total rhizoma coptidis on proliferation and apoptosis of human lung adenocarcinoma A549 cells and its possible mechanism. Methods: The A549 lung cancer cell lines were added with different concentrations of total flavonoids of Rhizoma Coptidis in vitro. The MTT assay was used to determine the inhibitory effect of total alkaloids of Rhizoma Coptidis (0.1,0.05,0.01,0.005 g · L -1) for 24 h, 48 h and 72 h, respectively. A549 cell growth inhibition. Hoechst staining was used to detect the growth of A549 cells treated with total alkaloids of Rhizoma Coptidis. Flow cytometry was used to detect the apoptosis induced by different concentrations of total alkaloids from I. The changes of mRNA expression of Caspase-3 and Survivin in A549 cells were detected by RT-PCR. Results: MTT results suggest that total alkaloids of Rhizoma Coptidis can inhibit the proliferation of lung cancer A549 cells in a dose-dependent manner. The results of MTT and Hoechst staining showed that compared with the negative group, the inhibitory effect on the proliferation of A549 cells was significantly inhibited by the total alkaloids group (P <0.05). The results of flow cytometry showed that the apoptosis rate of A549 cells was increased with the increase of drug concentration. The results of RT-PCR indicated that the ixoproliferative group up-regulated the expression of Caspase-3 mRNA and down-regulated the expression of Survivin mRNA. Conclusion: The total alkaloids of Rhizoma Coptidis can inhibit the proliferation and induce the apoptosis of human lung adenocarcinoma A549 cells in a time-dose relationship. The mechanism may be related to the up-regulation of Caspase-3 expression and down-regulation of Survivin expression.