吉祥草乙醇提取物Gracillin对肺癌A549细胞 侵袭、迁移及耐药的影响及机制研究

Effect and Mechanism of Gracillin from Reineckia carnea Ethanol Extract on Invasion, Migration and Drug Resistance of Lung Cancer A549 Cells

摘要: 目的：探讨吉祥草乙醇提取物活性成分Gracillin对肺癌A549细胞及其顺铂耐药株A549/DDP的侵袭、迁移及耐药性的影响及可能机制。方法：采用CCK8法测定Gracillin对A549及A549/DDP细胞的半数抑制浓度(IC50)；通过Transwell小室实验检测细胞侵袭和迁移能力；采用荧光定量PCR检测上皮间质转化(EMT)相关分子(E-cadherin、MMP2)及耐药相关分子(MDR1)的mRNA表达变化；通过联合用药及逆转耐药实验评估Gracillin与顺铂的协同作用及对耐药性的逆转效果。结果：Gracillin对A549和A549/DDP细胞的IC50分别为1.799 μmol/L和2.033 μmol/L，呈浓度依赖性抑制细胞增殖。Transwell实验表明，Gracillin显著抑制A549及A549/DDP细胞的侵袭和迁移能力(均P < 0.05)。Q-PCR结果显示，Gracillin可上调E-cadherin表达，下调MMP2及MDR1表达(均P < 0.05)。联合用药显示Gracillin与顺铂具有协同抑制A549细胞增殖的作用(CI < 1)，并能部分逆转A549/DDP细胞对顺铂的耐药性。结论：Gracillin在体外可抑制肺癌A549细胞及其顺铂耐药株的侵袭、迁移能力，增强耐药细胞对顺铂的敏感性，其机制可能与调控EMT相关分子及耐药相关分子MDR1的表达有关。

Abstract: Objective: To investigate the effect and mechanism of Gracillin, an active component from Reineckia carnea ethanol extract, on the invasion, migration and drug resistance of lung cancer A549 cells and cisplatin-resistant A549/DDP cells. Methods: CCK8 assay was used to determine the half-maximal inhibitory concentration (IC50) of Gracillin on A549 and A549/DDP cells. Transwell assay was performed to evaluate cell invasion and migration abilities. The mRNA expression levels of epithelial-mesenchymal transition (EMT)-related molecules (E-cadherin, MMP2) and drug resistance-related molecule (MDR1) were detected by quantitative real-time PCR. Combination index (CI) was calculated to assess the synergistic effect of Gracillin and cisplatin. Results: The IC50 values of Gracillin on A549 and A549/DDP cells were 1.799 μmol/L and 2.033 μmol/L, respectively, showing a concentration-dependent inhibitory effect on cell proliferation. Transwell assay showed that Gracillin significantly inhibited the invasion and migration abilities of both A549 and A549/DDP cells (all P < 0.05). Q-PCR results revealed that Gracillin upregulated E-cadherin expression while downregulating MMP2 and MDR1 expression (all P < 0.05). The combination of Gracillin and cisplatin exhibited synergistic effect on A549 cells (CI < 1), and Gracillin partially reversed the cisplatin resistance of A549/DDP cells. Conclusion: In vitro Gracillin can inhibit the invasion and migration abilities of lung cancer A549 cells and cisplatin-resistant A549/DDP cells, and enhance the sensitivity of resistant cells to cisplatin. The mechanism may involve the regulation of EMT-related molecules and the drug resistance-related molecule MDR1.