论文部分内容阅读
目的:培养恶性腹膜间皮瘤(MPM)细胞，观察阿帕替尼对其生物学特性影响。方法:通过MPM手术标本建立裸小鼠模型，于2019年8月至10月利用此模型培养MPM细胞，采用瑞士-吉姆萨及免疫细胞化学染色鉴定；实验分阿帕替尼组、溶剂组及空白对照组；细胞计数试剂盒8(CCK-8)、流式细胞术、划痕及半胱天冬酶3(Caspase-3)活性检测法研究对细胞增生、周期、迁移及凋亡影响；组间比较采用单因素方差分析及n t检验。n 结果:培养MPM细胞，钙调蛋白(Calretinin)、细胞角蛋白(CK)5/6、p53、细胞增殖核抗原-67(Ki-67)阳性；0、12.5、25.0、50.0、100.0 μmol/L药物处理24 h，细胞活力100%、(83.78±5.58)%、(68.03±2.20)%、(47.31±1.03)%、(38.47±6.68)%，与对照组比较，不同浓度阿帕替尼均抑制细胞活力(n F＝118.000、5.031、25.120、88.600、15.950，n P<0.01)，差异有统计学意义；划痕实验空白、溶剂、25 μmol/L及50 μmol/L组细胞迁移率(77.52±2.54)%、(78.50±6.91)%、(26.43±15.27)%和(13.03±11.93)%，与对照组比较，不同浓度阿帕替尼均抑制细胞迁移(n F＝64.920，n t＝8.084，7.609，12.950，11.630，n P<0.01)，差异有统计学意义；药物抑制Gn 2/M期但不诱导凋亡。n 结论:阿帕替尼抑制MPM细胞增生及迁移，影响细胞周期。“,”Objective:To establish primary cells of malignant peritoneal mesothelioma (MPM), and study the effect of apatinib.Methods:MPM surgical specimens were used to establish nude mice models, and then the models were used to obtain MPM cells from August to October 2019, which were identified by Swiss-Gimsa and immunocytochemical staining. Experiment was divided into apatinib group, solvent group and blank control group. Cell counting kit-8 (CCK-8), flow cytometry, wound-healing, and Caspase-3 assays were performed, respectively, to study effects on proliferation, cycle, movement and apoptosis. GraphPad Prism 8.0 was used for statistical analysis. Statistical methods were one-way ANOVA and n t test.n Results:MPM cells were cultured. Calretinin, CK5/6, p53, and proliferation cell nuclear antigen (Ki-67) staining was positive. After treatment with 0, 12.5, 25.0, 50.0, and 100 μmol/L apatinib for 24 h, cell viability was 100%, (83.78±5.58)%, (68.03±2.20)%, (47.3±1.03)% and (38.47±6.68)% respectively ( n F=118.000, n P<0.01; 12.5 μmol/L vs. controln t=5.031, n P<0.01; 25 μmol/L vs. controln t=25.120, n P<0.01; 50 μmol/L vs. controln t=88.600, n P<0.01; 100 μmol/L vs. controln t=15.950, n P<0.01); Wound-healing showed that migration rates of blank, solvent, 25 and 50 μmol/L apatinib groups were (77.52±2.54)%, (78.50±6.91)%, (26.43±15.27)%, and (13.03±11.93)% respectively (n F=64.920, n P<0.01; 25 μmol/L vs. blankn t=8.084, n P<0.01; 25 μmol/L vs. solventn t=7.609, n P<0.01; 50 μmol/L vs. blankn t=12.950, n P<0.01; 50 μmol/L vs. solventn t=11.630, n P<0.01); Apatinib could inhibit growth of cells in Gn 2/M phase, but not induce apoptosis.n Conclusion:Apatinib can inhibit proliferation, migration and cycle of MPM cells, but not induce apoptosis.