高压氧联合多柔比星处理对人低分化胃癌MKN45细胞增殖和凋亡的影响  

Hyperbaric oxygen combined with doxorubicin on the proliferation and apoptosis of human gastric cancer MKN45 cells

作　　者：李文龙[1] 高伟[2] 王全兵 张丽娜[4] 梁宏伟[5] Li Wenlong;Gao Wei;Wang Quanbing;Zhang Lina;Liang Hongwei(Department of General Surgery,Second Hospital of Shanxi Medical University,Taiyuan 030001,China;Department of Gastroenterology,Second Hospital of Shanxi Medical University,Taiyuan 030001,China;Rehabilitation Department,Shanxi Bethune Hospital,Taiyuan 030032,China;Department of General Practice,Taiyuan 030001,China;Department of Nuclear Medicine,Taiyuan 030001,China)

机构地区：[1]山西医科大学第二医院普通外科,太原030001 [2]山西医科大学第二医院消化内科,太原030001 [3]白求恩医院康复科,太原030032 [4]山西医科大学第二医院全科医学科,太原030001 [5]山西医科大学第二医院核医学科,太原030001

出　　处：《中华航海医学与高气压医学杂志》2022年第6期813-817,共5页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

摘　　要：目的:探讨高压氧(HBO)联合多柔比星(DOX)对人低分化胃癌MKN45细胞增殖和凋亡的影响及其作用机制。方法:体外培养人胃癌MKN45细胞完成后,按照实验设计将细胞分为4组:对照组、DOX组、HBO组和HBO+DOX组。对照组加入DMSO培养基,DOX组加入终质量浓度为2 mg/L DOX溶液;HBO组则在高压氧舱(0.25 MPa)中暴露培养90 min,HBO+DOX组先在高压氧舱暴露培养90 min后再用2 mg/L DOX溶液处理。利用CCK-8法检测HBO+DOX处理对胃癌MKN45细胞的增殖抑制作用,利用流式细胞术检测胃癌MKN45细胞的凋亡率,Western blotting实验检测胃癌MKN45细胞内凋亡蛋白及JNK/STAT3信号通路相关蛋白的表达水平。结果:HBO+DOX组MKN45细胞存活率明显低于对照组和HBO组(P<0.01),也低于PTX组(P<0.05)。HBO+DOX组MKN45细胞凋亡率明显高于对照组和HBO组(P<0.01),也高于PTX组(P<0.05)。与对照组和HBO组比较,DOX组和HBO+DOX组MKN45细胞内Bad、caspase-3、PARP蛋白表达明显升高,Bcl-2蛋白表达明显降低(P<0.01);DOX组PARP蛋白表达明显低于HBO+DOX组(P<0.05)。与对照组和HBO组比较,DOX组和HBO+DOX组MKN45细胞内p-JNK蛋白表达明显降低,且HBO+DOX组低于DOX组(P<0.05或P<0.01)。与对照组比较,DOX组和HBO+DOX组MKN45细胞内p-STAT3蛋白表达明显升高(P<0.05)。与对照组、HBO组和DOX组比较,HBO+DOX组MKN45细胞内耐药性相关蛋白P-gp明显降低(P<0.01)。结论:HBO处理能够增强DOX对人胃癌MKN45细胞的增殖抑制作用及促凋亡作用,其作用机制可能是通过调节MKN45细胞中的JNK/STAT3信号通路来实现的。Objective:To explore the effect of hyperbaric oxygen(HBO)combined with doxorubicin(DOX)on the proliferation and apoptosis of human gastric cancer MKN45 cell line,the poorly differentiated adenocarcinoma cells,and its associated mechanism.Methods:After culturing human gastric cancer MKN45 cells in vitro,the cells were divided into four groups according to the experimental design:control group,DOX group,HBO group,and HBO+DOX group.The control group was given DMSO,and the DOX group was given DOX solution with a final concentration of 2 mg/L.The HBO group was exposed in the hyperbaric oxygen chamber(0.25 MPa)for 90 min,while the HBO+DOX group was given 2 mg/L of DOX after being exposed in a hyperbaric oxygen chamber for 90 min.The CCK-8 method was used to detect the inhibitory effect of HBO+DOX treatment on the proliferation of gastric cancer MKN45 cells;the flow cytometry was used to detect the apoptosis rate of gastric cancer MKN45 cells;and the western blotting was used to detect the expression levels of apoptosis proteins and JNK/STAT3 signaling pathway related proteins in gastric cancer MKN45 cells.Results:The survival rate of MKN45 cells in the HBO+DOX group was significantly lower than those in the control group,the HBO group(P<0.01),and in the PTX group(P<0.05).The apoptosis rate of MKN45 cells in the HBO+DOX group was significantly higher than those in the control group,the HBO group(P<0.01),and the PTX group(P<0.05).Compared with the control group and the HBO group,the expression levels of Bad,caspase-3,and PARP protein in MKN45 cells in the DOX group and the HBO+DOX group were significantly increased,while the expression levels of Bcl-2 protein were significantly decreased(P<0.01).PARP protein expression level in the DOX group was significantly lower than that in the HBO+DOX group(P<0.05).Compared with the control group and the HBO group,p-JNK protein expression levels in MKN45 cells in the DOX group and the HBO+DOX group decreased significantly,and the p-JNK protein expression level in the HBO+DOX group w

关 键 词：高压氧 多柔比星 胃癌 MKN45细胞 细胞凋亡 

分 类 号：R735.2[医药卫生—肿瘤]