Amorphigenin联合顺铂对人肺腺癌A549/DDP细胞的协同抗肿瘤作用

Synergistic Antitumor Effect of Amorphigenin Combined with Cisplatin in Human Lung Adenocarcinoma A549/DDP Cells

背景与目的 Amorphigenin是从紫穗槐属植物的种子中分离提取的鱼藤酮类化合物,研究发现amorphigenin对多种肿瘤细胞具有增殖抑制作用。本研究拟探讨amorphigenin对人肺腺癌耐顺铂细胞株A549/DDP的抗肿瘤作用及其可能的分子机制。方法采用CCK-8法测定A549/DDP细胞的增殖;克隆形成实验测定A549/DDP细胞的克隆形成;流式细胞术检测细胞的凋亡率;Western blot技术检测caspase-3、PARP和LRP蛋白的表达。结果 Amorphigenin可抑制A549/DDP细胞的增殖48 h[半数抑制浓度(half maximal inhibitory concentration,IC_(50))]为(2.19±0.92)μmol/L、抑制克隆形成及诱导细胞凋亡。此外,Amorphigenin与顺铂联合可协同地抑制A549/DDP细胞生长和促进凋亡;降低耐药蛋白LRP蛋白的表达。结论 Amorphigenin可抑制A549/DDP细胞增殖和促进细胞凋亡;amorphigenin可能是通过抑制耐药蛋白LRP蛋白表达,进而与顺铂对A549/DDP细胞产生协同抑制作用。

Background and objective Amorphigenin, a rotenoid compouns, from seeds ofAmorpha fruticosa, has been shown to possess anti-proliferation activities in several cancer cells. To explore the antitumor effects of amorphigenin on cisplatin-resistant human lung adenocarcinomaA549/DDP cells and explore the underlying mechanisms. Methods CCK-8 assay was used to measure the proliferation of A549/DDP cells; Colony formation assay was used to measure the colony formation of A549/DDP cells; Flow cytometry assay was used to detect the apoptosis rates; Western blot analysis was used to explore the expression of apoptosis-related proteins （caspase-3 protein, PARP protein） and lung resistance protein （LRP）. Results Our results demonstrated that amorphigenin could inhibit the proliferation of A549/DDP cells with a inhibition con- centration of 50% cell growth （ICs0） at 48 h of （2.19±0.92） μmol/L. Amorphigenin could inhibit the colony formation ability and induce apoptosis of A549/DDP cells; Furthermore, amorphigenin combined with cisplatin showed synergistic prolifera- tion-inhibitory effect and apoptosis-promoting effect in A549/DDP cells; reduced the expression ofLRP of A549/DDP cells. Conclusion Amorphigenin remarkably inhibits the proliferation and induces apoptosis in A549/DDP cells. Combination of amorphigenin with cisplatin had the synergistic inhibitory effect on A549/DDP cells by downregulating the expression of LRP.