论文部分内容阅读
目的:探讨人胎盘提取物(human placenta extracts,HPE)对人结肠癌细胞株Caco-2及SW480增殖和迁移能力的影响。方法:用不同质量浓度HPE处理Caco-2和SW480细胞,CCK-8、Ki-67、CFSE法检测细胞增殖活性的变化,Annexin-V/PI及DAPI细胞核染色检测HPE对细胞周期及凋亡的影响,实时荧光定量PCR法检测HPE对细胞中BAX、CDK2及Cyclin A2基因表达的影响,划痕实验检测对细胞迁移能力的影响。建立裸鼠结肠癌移植瘤模型,随机分组分别单独或联合给予HPE和5-氟尿嘧啶(5-Fu),观察裸鼠移植瘤的生长状况。结果:HPE处理组Caco-2及SW480细胞增殖能力低于对照组[(0.82±0.01)vs(0.96±0.02),P<0.01;(0.90±0.03)vs(0.96±0.02),P<0.05],细胞凋亡率高于对照组[(20.47±1.32)%vs(11.01±3.82)%,P<0.01;(20.70±5.19)%vs(8.00±2.69)%,P<0.05];HPE处理组细胞胞质减少、核固缩、BAX基因表达增加[(3.23±1.90)vs(1.00±0.00),(2.25±0.55)vs(1.00±0.00),均P<0.05];细胞停留在细胞DNA复制S期,出现凋亡峰;Caco-2细胞CDK2及Cyclin A2基因表达降低(P<0.05);并且细胞的迁移能力低于对照组[(0.17±0.29)vs(1.50±0.50)mm,P<0.05]。HPE处理组裸鼠移植瘤体积小于对照组、生存率高于单用5-Fu组(P<0.05)。结论:HPE在体内外通过干预结肠癌细胞凋亡、阻滞细胞周期、抑制细胞增殖及迁移,可能对抑制肿瘤细胞增殖有一定作用。
Objective: To investigate the effect of human placenta extracts (HPE) on the proliferation and migration of human colon cancer cell lines Caco-2 and SW480. Methods: Caco-2 and SW480 cells were treated with different concentrations of HPE, and the changes of cell proliferation were detected by CCK-8, Ki-67 and CFSE. The cell cycle and apoptosis were detected by nuclear staining of Annexin-V / PI and DAPI The effect of HPE on the expression of BAX, CDK2 and Cyclin A2 in cells was detected by real-time fluorescence quantitative PCR, and the influence of scratch test on cell migration was assayed. The model of colon cancer xenografts in nude mice was established. HPE and 5-fluorouracil (5-Fu) were administered separately or in combination to observe the growth of xenografts in nude mice. RESULTS: The proliferation of Caco-2 and SW480 cells in HPE-treated group was significantly lower than that in control group [(0.82 ± 0.01) vs (0.96 ± 0.02), P <0.01; 0.90 ± 0.03 vs 0.96 ± 0.02, P <0.05] (20.47 ± 1.32)% vs (11.01 ± 3.82)%, P <0.01; (20.70 ± 5.19)% vs (8.00 ± 2.69)%, P <0.05] (3.23 ± 1.90) vs (1.00 ± 0.00), (2.25 ± 0.55) vs (1.00 ± 0.00), all P <0.05]; cells stayed in the cell DNA replication (P <0.05). The migration of cells in Caco-2 cells was lower than that in control group [(0.17 ± 0.29) vs (1.50 ± 0.50) mm, P < 0.05]. The volume of xenografted tumor in HPE-treated group was smaller than that in control group, and the survival rate was higher than that of 5-Fu alone group (P <0.05). Conclusion: HPE can inhibit the proliferation of tumor cells in vitro and in vivo by interfering with the apoptosis of colon cancer cells, arresting the cell cycle and inhibiting cell proliferation and migration.