金复康优化方联合吉非替尼对非小细胞肺癌细胞H1975的影响及作用机制(1)
【摘 要】 目的：考察金復康优化方(ALG-X-012)联合吉非替尼对非小细胞肺癌H1975的增殖影响，探究其可能的作用机制。方法：通过MTT法、划痕实验、流式细胞术实验考察ALG-X-012、吉非替尼单独用药以及两者联合用药对H1975细胞的增殖抑制、细胞迁移与凋亡的影响；Western blot法检测两者单独用药及联合用药对 p-EGFR、p-AKT、p-mTOR相关蛋白表达的影响。结果：MTT结果显示低于5μmol/L浓度吉非替尼对H1975细胞增殖无抑制作用，但与ALG-X-012联用给药后，对H1975细胞即表现出明显抑制作用。划痕实验结果表明与1μmol/L吉非替尼单独用药相比，联用ALG-X-012可明显抑制H1975细胞迁移。流式细胞术实验结果显示，ALG-X-012、吉非替尼及两者联合用药干预H1975细胞后凋亡率分别是(3.93±0.15)%、(4.47±0.28)%与(11.43±0.96)%，联合用药可显著提高吉非替尼诱导细胞凋亡率(P<0.01)。Western blot结果显示，吉非替尼组和ALG-X-012组对各项蛋白指标均无明显影响，联合用药组却能明显下调H1975细胞p-EGFR、p-AKT、p-mTOR蛋白表达。结论：ALG-X-012联合低浓度吉非替尼即可显著抑制H1975细胞的增殖，诱导肿瘤细胞凋亡，抑制肿瘤细胞迁移，其抗肿瘤活性的机制可能与阻断EGFR-PI3k-AKT信号通路有关。
【关键词】 ALG-X-012；吉非替尼；非小细胞肺癌；表皮生长因子受体酪氨酸激酶抑制剂；EGFR-PI3K-AKT信号传导通路
【中图分类号】R-33 【文献标志码】 A 【文章编号】1007-8517(2018)07-0021-06
Abstract：Objective To observe the effects of optimized ALG-X-012 combined gefitinib on non small cell lung cancer H1975 growth， to explore its possible mechanism. Methods The MTT assay， scratch assay， flow cytometry， ALG-X-012 experiment to observe the effects of gefitinib monotherapy and combination of both on H1975 cell proliferation inhibition， cell migration and apoptosis； The expression of p-mTOR，p-EGFR and p-AKT of monotherapy and combination treatment were detected by Western blot.Results The results of MTT showed that gefitinib did not inhibit the growth of H1975 cells when the concentration was less than 5μmol/L， but it was significantly inhibited by H1975 combined with ALG-X-012. The scratch test showed that compared with 1μmol/L gefitinib， the combination of ALG-X-012 with gefitinib could significantly inhibit the migration of H1975 cells. Flow cytometry results showed that ALG-X-012， gefitinib and combination intervention after H1975 cell apoptosis rate were (3.93+0.15)%， (4.47+0.28)% and (11.43+0.96)%， combination therapy can significantly improve gefitinib induced apoptosis rate (P<0.01). Western blot results showed that the gefitinib group and ALG-X-012 group had no significant effect on the protein indexes， while the combination group could significantly reduce the expression of p-EGFR， p-AKT， p-mTOR and p-Met protein in H1975 cells. Conclusion ALG-X-012 combined with gefitinib can significantly inhibit the proliferation of H1975 cells， induce tumor cell apoptosis and inhibit tumor cell migration. The mechanism of its antitumor activity may be related to blocking EGFR-PI3k-AKT signaling pathway.
Keywords：ALG-X-012； Gefitinib； Non-small Cell Lung Cancer； Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors； EGFR-PI3K-AKT Signaling Pathway, 百拇医药(杜梅 杨蕾 刘丹)
【关键词】 ALG-X-012；吉非替尼；非小细胞肺癌；表皮生长因子受体酪氨酸激酶抑制剂；EGFR-PI3K-AKT信号传导通路
【中图分类号】R-33 【文献标志码】 A 【文章编号】1007-8517(2018)07-0021-06
Abstract：Objective To observe the effects of optimized ALG-X-012 combined gefitinib on non small cell lung cancer H1975 growth， to explore its possible mechanism. Methods The MTT assay， scratch assay， flow cytometry， ALG-X-012 experiment to observe the effects of gefitinib monotherapy and combination of both on H1975 cell proliferation inhibition， cell migration and apoptosis； The expression of p-mTOR，p-EGFR and p-AKT of monotherapy and combination treatment were detected by Western blot.Results The results of MTT showed that gefitinib did not inhibit the growth of H1975 cells when the concentration was less than 5μmol/L， but it was significantly inhibited by H1975 combined with ALG-X-012. The scratch test showed that compared with 1μmol/L gefitinib， the combination of ALG-X-012 with gefitinib could significantly inhibit the migration of H1975 cells. Flow cytometry results showed that ALG-X-012， gefitinib and combination intervention after H1975 cell apoptosis rate were (3.93+0.15)%， (4.47+0.28)% and (11.43+0.96)%， combination therapy can significantly improve gefitinib induced apoptosis rate (P<0.01). Western blot results showed that the gefitinib group and ALG-X-012 group had no significant effect on the protein indexes， while the combination group could significantly reduce the expression of p-EGFR， p-AKT， p-mTOR and p-Met protein in H1975 cells. Conclusion ALG-X-012 combined with gefitinib can significantly inhibit the proliferation of H1975 cells， induce tumor cell apoptosis and inhibit tumor cell migration. The mechanism of its antitumor activity may be related to blocking EGFR-PI3k-AKT signaling pathway.
Keywords：ALG-X-012； Gefitinib； Non-small Cell Lung Cancer； Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitors； EGFR-PI3K-AKT Signaling Pathway, 百拇医药(杜梅 杨蕾 刘丹)