异氟醚对人结肠癌细胞SW480 生长、侵袭和迁移能力的调节作用
| 作者: |
1任伟荣,
1王丽,
1薛利军
1 榆林市第一医院手术麻醉科，陕西 榆林 719000 |
| 通讯: |
任伟荣
Email: wk_87815@126.com |
| DOI: | 10.3978/j.issn.2095-6959.2018.07.001 |
| 基金: | 陕西省科技计划项目 (2014jh-20)。 |
摘要
目的：探究异氟醚对人结肠癌S W480细胞增殖、凋亡、侵袭和迁移的影响及其分子机制。 方法：SW480细胞随机分为对照组(SW480)、异氟醚(1.5%，2.0%，2.5%)组，共4组。噻唑蓝比色法(MTT)检测细胞增殖，Hoechst染色检测细胞凋亡，Transwell检测细胞侵袭，划痕试验分析细胞迁移，Western印迹法检测细胞增殖核抗原67(antigen identified by monoclonal antibody，Ki-67)， Caspase-3，LC3II，LC3I，血管内皮细胞生长因子(vascular endothelial growth factor，VEGF)，磷脂酰肌醇3-激酶(phosphatidylinositide 3-kinases，PI3K)，p-PI3K，蛋白激酶B(protein kinase B， AKT)和p-AKT蛋白水平。结果：与对照组相比，异氟醚(1.5%，2.0%，2.5%)组细胞增殖倍数下降(P=0.0093，n=5)。异氟醚(1.5%，2%，2.5%)组细胞凋亡率高于对照组(P=0.0087，n=5)。而且，异氟醚(1.5%，2.0%，2.5%)组每个视野下的侵袭细胞数低于对照组(P=0.0081，n=5)。与对照组相比，异氟醚(1.5%，2.0%，2.5%)组划痕愈合率降低(P=0.0078，n=5)。异氟醚(1.5%， 2.0%，2.5%)组Ki-67和VEGF及LC3II/LC3I的比值表达低于对照组，Caspase-3表达高于对照组 (P=0.0096，n=5)。另外，异氟醚(1.5%，2.0%，2.5%)组p-PI3K/PI3K和p-AKT/AKT比值低于对照组(P=0.0099，n=5)。IGF-1可逆转异氟醚诱导的p-PI3K，p-AKT，Ki-67及VEGF蛋白水平升高和Caspase的蛋白水平下降(P=0.0079，n=5)。结论：异氟醚可通过抑制PI3K/AKT信号通路活化减弱人结肠癌细胞SW480细胞增殖、侵袭和迁移，促进细胞凋亡。
关键词：
异氟醚；结肠癌；增殖；凋亡；侵袭；迁移
Effects of isoflurane on the cell growth, invasion and migration of colon cancer SW480 cells
CorrespondingAuthor: REN Weirong Email: wk_87815@126.com
DOI: 10.3978/j.issn.2095-6959.2018.07.001
Foundation: This work was supported by Shaanxi Science and Technology Project Foundation, China (2014jh-20).
Abstract
Objective: Colorectal cancer is the third most common cancer in the world. This study aims to explore the effects of isoflurane on the cell proliferation, apoptosis, invasion and migration of colon cancer SW480 cells. Methods: SW480 cells were randomly divided into four groups, including control group (SW480), isoflurane (1.5%, 2.0%, 2.5%) group. Cell proliferation was tested by MTT, apoptosis was measured by Hoechst staining, cell invasion was detected by transwell, cell migration was measured by wound healing. The protein levels of antigen identified by monoclonal antibody (Ki-67), Caspase-3, LC3II, LC3I, vascular endothelial growth factor (VEGF), phosphatidylinositide 3-kinases (PI3K), p-PI3K, protein kinase B (AKT) and p-AKT were detected by Western blot. Results: Compared with the control group, the proliferation in the isoflurane (1.5%, 2.0%, 2.5%) group was reduced (P=0.0093, n=5). The apoptosis rate in the isoflurane (1.5%, 2.0%, 2.5%) group was higher than control group (P=0.0087, n=5). The number of invasive cells per field in the isoflurane (1.5%, 2.0%, 2.5%) group was lower than that in the control group (P=0.0081, n=5). Compared with the control group, the wound closure rate in the isoflurane (1.5%, 2.0%, 2.5%) group decreased (P=0.0078, n=5). Compared with the control group, the expression of Ki-67 and VEGF and rate of LC3II/LC3I in the isoflurane (1.5%, 2.0%, 2.5%) group reduced with enhancive expression of Caspase-3 (P=0.0096, n=5). In addition, the rate of p-PI3K/PI3K and p-AKT/AKT in the isoflurane (1.5%, 2.0%, 2.5%) group was lower than that in the control group (P=0.0099, n=5). The elevated protein levels of p-PI3K, p-AKT, Ki-67 and VEGF and declined protein levels of Caspase-3 by isoflurane was reversed by IGF-1 (P=0.0079, n=5). Conclusion: Isoflurane alleviates the proliferation, invasion, migration and promotes apoptosis of colon cancer SW480 cells via inhibiting activation of PI3K/AKT pathway.
Keywords：
isoflurane; colon cancer; proliferation; apoptosis; invasion; migration