蛇葡萄素钠联合卡铂对人肺腺癌GLC-82细胞增殖及凋亡的影响

Effect of ampelopsin sodium combined with carboplatin on the proliferation and apoptosis of human lung adenocarcinoma cell GLC-82

目的探讨蛇葡萄素钠(AMP-Na)单用及其与卡铂(CBP)合用对人肺腺癌GLC-82细胞增殖的抑制作用及可能的机制。方法四甲基偶氮唑盐(MTT)比色法测定两药单用及合用对GLC-82细胞的体外杀伤活性;透射电子显微镜(TEM)观察GLC-82细胞超微结构的变化;流式细胞仪(FCM)检测GLC-82细胞凋亡和caspase-3的表达。结果对于GLC-82细胞,AMP-Na与CBP合用,CBP的IC50由(17.10±4.78)mg·L-1降低到<3.12 mg·L-1(P<0.01),表明Amp-Na对卡铂抑制GLC-82细胞增殖的作用具有协同效应(CDI<1)。TEM及FCM检测结果表明,单用AMP-Na及与卡铂合用均可诱导GLC-82细胞凋亡、坏死,两药合用后坏死率由(2.56±0.41)%升高到(71.83±5.43)%(P<0.01)。AMP-Na单用及与卡铂合用作用于GLC-82细胞48 h后,caspase-3的表达均明显增高。结论AMP-Na和CBP具有协同诱导GLC-82细胞凋亡的作用,机制可能与激活细胞内caspase-3介导的信号转导通路相关。

Aim To investigate the cytotoxic effect and mechanism of ampelopsin sodium （ AMP-Na ） on hu-man lung adenocarcinoma cell line GLC-82 alone or combined with carboplatin （ CBP ） . Methods The cytotoxic effect of human lung adenocarcinoma cell line GLC-82 was investigated by 3-（4,5-dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide （ MTT ） colori-metric assay. Ultrastructure change of apoptotic GLC-82 cells was observed with transmission electron micro-scope. The changes of the cell apoptosis and the ex-pression of caspase-3 were analyzed with flow cytome-ter. Results Combined with AMP-Na, the IC50 of CBP decreased from （17. 10 ± 4. 78） mg·L^-1 to 〈3. 12 mg·L^-1（P〈0. 01）, showed that the combina-tion of AMP-Na and CBP had synergistic effect on GLC-82 cells （ CDI 〈1 ） . As with transmission electron microscope and flow cytometric analysis, the apop-tosis and necrosis ratios also increased in the combina-tion group. The necrosis ratios increased from （2. 56 ± 0. 41 ）% to （ 71. 83 ± 5. 43 ）% （ P〈0. 01 ） . The ex-pression of caspase-3 was increased significantly after treated with AMP-Na or combined with CBP. Conclu-sions There is a synergistic cytotoxic effect on GLC-82 cells treated with AMP-Na combined with CBP. Ap-optotic cells and necrotic cells are found in GLC-82 cells treated with AMP-Na alone or combined with CBP. One of the mechanisms to induce apoptosis is probably that activation of caspase-3 mediates signal transduction pathway in cells.