摘要
目的探讨气体麻醉剂七氟醚影响骨肉瘤细胞侵袭转移能力及化疗敏感性的分子机制。方法选择骨肉瘤MG63细胞进行体外培养,分别使用气体麻醉剂七氟醚和化疗药物顺铂处理细胞。将细胞分为正常对照组(不进行任何处理)、七氟醚组(5.0%的七氟醚干预4 h)、顺铂组(终浓度10 mg/L的顺铂干预)和七氟醚+顺铂组(终浓度10 mg/L的顺铂+5.0%的七氟醚干预4 h)。采用MTT法测定七氟醚对骨肉瘤MG63细胞增殖能力的影响;采用流式细胞仪测定七氟醚对骨肉瘤MG63细胞凋亡情况的影响;采用Trans well小室法和划痕实验分别检测各组细胞的侵袭和迁移能力;采用蛋白质印迹(Western Blotting)法测定与侵袭转移相关的E-cadherin蛋白表达,以及相关耐药蛋白P-gp、Bcl-2和p53的表达情况。结果不同浓度(2.5%~20.0%)七氟醚干预后,骨肉瘤MG63细胞增殖能力显著降低,并且5.0%的七氟醚作用效果最显著,抑制率为(14.18±0.64%),差异有统计学意义(P<0.05)。与正常对照组相比,七氟醚组中MG63细胞的侵袭细胞数(126.28±11.95个vs.73.46±8.66个)和细胞迁移率(76.63±8.93 vs.46.79±4.52)显著降低,E-cadherin蛋白表达量(0.13±0.03 vs.0.36±0.07)显著升高,差异有统计学意义(P<0.05)。与正常对照组和七氟醚组相比,顺铂组和七氟醚+顺铂组细胞凋亡率和p53显著升高,P-gp和Bcl-2显著降低(P<0.05),而前2组以上指标相比差异均无统计学意义(P>0.05);而与顺铂组相比,七氟醚+顺铂组细胞凋亡率[(54.52±5.74)%vs.(35.43±4.65)%]和p53(0.12±0.03 vs.0.49±0.09)显著降低,P-gp(0.25±0.07 vs.0.79±0.11)和Bcl-2(1.34±0.13 vs.0.89±0.08)显著升高,差异有统计学意义(P<0.05)。结论七氟醚可能通过提高E-cadherin蛋白表达量抑制骨肉瘤细胞的增殖和侵袭转移能力,同时可通过促进P-gp和Bcl-2的表达以及抑制p53的表达降低MG63细胞对化疗药物的敏感性。
Objective In this study,the molecular mechanism of gaseous anaesthetic affects the invasion and metastasis ability and the sensitivity to chemotherapy of osteosarcoma cells was investigated.Methods The MG63 cells of osteosarcoma were cultured in vitro and treated with sevoflurane and cisplatin respectively.The cells were divided into normal control group(no treatment),sevoflurane group(5.0%sevoflurane intervention for 4 h),cisplatin group(cisplatin intervention with a final concentration of 10 mg/L),and sevoflurane+cisplatin group(final concentration of 10 mg/L cisplatin+5.0%sevoflurane intervention for 4 h).The effect of sevoflurane on proliferation of osteosarcoma MG63 cells was determined by MTT assay.The effect of sevoflurane on apoptosis of osteosarcoma MG63 cells was determined by flow cytometry.The invasion and migration ability of each group of cells were detected by Trans well small chamber method and scratch test respectively.Western Blot was used to determine the expression of E-cadherin protein related to invasion and metastasis,as well as the expressions of related drug-resistant proteins P-gp,Bcl-2 and p53.Results After the intervention of different concentrations(2.5%~20.0%)of sevoflurane,the proliferation ability of osteosarcoma MG63 cells was significantly reduced,and the effect of 5.0%sevoflurane was the most significant,and the inhibition rate was(14.18±0.64)%,the difference was statistically significant(P<0.05).Compared with the normal control group,the number of invasive cells(126.28±11.95 vs.73.46±8.66)and cell migration rate(76.63±8.93 vs.46.79±4.52)of MG63 cells in the sevoflurane group were significantly reduced,the expression of E-cadherin protein(0.13±0.03 vs.0.36±0.07)was significantly increased,the difference was statistically significant(P<0.05).Compared with the normal control group and sevoflurane group,the apoptosis rate and p53 of the cisplatin group and the sevoflurane+cisplatin group were significantly increased,and P-gp and Bcl-2 were significantly reduced(P<0.05),but there was no statistically significant difference in the above indicators between the first two groups(P>0.05);compared with the cisplatin group,the apoptotic rate[(54.52±5.74)%vs.(35.43±4.65)%]and p53(0.12±0.03 vs.0.49±0.09)of the sevoflurane+cisplatin group were significantly reduced,and P-gp(0.25±0.07 vs.0.79±0.11)and Bcl-2(1.34±0.13 vs.0.89±0.08)were significantly increased,the differences were statistically significant(P<0.05).Conclusion Sevoflurane may inhibit the proliferation,invasion and metastasis of osteosarcoma cells by increasing the expression of E-cadherin protein,and at the same time,it can reduce the sensitivity of MG63 cells to chemotherapy drugs by promoting the expression of P-gp and Bcl-2 and inhibiting the expression of p53.
作者
鲁飞
王强
谷淼
LU Fei;WANG Qiang;GU Miao(Department of Anesthesiology,Fuxin Central Hospital,Fuxin Liaoning 123000,China;Department of Anesthesiology,Shenyang Chest Hospital,Shenyang Liaoning 110000,China;Department of Anesthesiology,Third Affiliated Hospital of Jinzhou Medical University,Jinzhou Liaoning 121001,China)
出处
《临床和实验医学杂志》
2020年第24期2614-2618,共5页
Journal of Clinical and Experimental Medicine
基金
辽宁省自然基金资助计划(编号:2019-MS-134)。
关键词
气体麻醉剂
七氟醚
骨肉瘤细胞
化疗敏感性
Gaseous anaesthetic
Sevoflurane
Osteosarcoma cells
Chemo-sensitivity
