摘要
目的探讨过氧化氢(H_(2)O_(2))联合透明质酸钠是否可以提高宫颈癌细胞的放疗敏感性,并探讨其潜在机制。方法将细胞分为对照组、透明质酸钠组、4 Gy组、透明质酸钠+4 Gy组,Western blotting实验检测自噬相关蛋白水平。将细胞分为对照组,透明质酸钠组,200μmol/L H_(2)O_(2)组、500μmol/L H_(2)O_(2)组、700μmol/L H_(2)O_(2)组和1000μmol/L H_(2)O_(2)组,CCK8实验检测H_(2)O_(2)对细胞活力的影响;将细胞分为对照组和200μmol/L H_(2)O_(2)组,分别进行0、2、4、6、8 Gy放疗,克隆形成实验评估H_(2)O_(2)的放疗增敏作用。将细胞分为对照组、200μmol/L H_(2)O_(2)组、4 Gy组、200μmol/L H_(2)O_(2)+4 Gy组,Western blotting实验和免疫荧光实验检测自噬相关蛋白水平。结果对照组与透明质酸钠组的自噬相关蛋白LC3 II、p62表达差异无统计学意义(P>0.05);4 Gy组与透明质酸钠+4 Gy组的自噬相关蛋白LC3 II、p62表达差异无统计学意义(P>0.05)。CCK8实验结果表明,放疗前后,对照组和透明质酸钠组的细胞增殖率差异无统计学意义(P>0.05);与对照组相比,200μmol/L H_(2)O_(2)组促进细胞增殖(P<0.05),500μmol/L H_(2)O_(2)组对细胞增殖的影响无统计学意义(P>0.05),700μmol/L H_(2)O_(2)组和1000μmol/L H_(2)O_(2)组抑制细胞增殖(P<0.05)。克隆形成实验结果表明,H_(2)O_(2)增加了细胞对放疗的敏感性(P<0.05)。Western blotting实验和免疫荧光实验表明,与对照组相比,200μmol/L H_(2)O_(2)组抑制了自噬(P<0.05);与4 Gy组相比,200μmol/L H_(2)O_(2)+4 Gy组促进了自噬(P<0.05)。与对照组相比,200μmol/L H_(2)O_(2)组p-AKT和p-mTOR蛋白表达增加(P<0.05);与4 Gy组相比,200μmol/L H_(2)O_(2)+4 Gy组p-AKT和p-mTOR的表达减少(P<0.05)。结论H_(2)O_(2)通过调控AKT/mTOR通路进而调控自噬来提高宫颈癌的放疗敏感性。
Objective To investigate whether hydrogen peroxide,represented as H_(2)O_(2),combined with sodium hyaluronate can improve the sensitivity of cervical cancer cells to radiotherapy and to explore the underlying mechanisms.Methods The cells were divided into control group,sodium hyaluronate group,4 Gy group,sodium hyaluronate+4 Gy group,and the expressions of autophagy related proteins were detected with Western blotting assay.Then,the cells were divided into control group,sodium hyaluronate group,200μmol/L H_(2)O_(2) group,500μmol/L H_(2)O_(2) group,700μmol/L H_(2)O_(2) group and 1000μmol/L H_(2)O_(2) group,and CCK8 experiment was used to detect the effect of H_(2)O_(2) on cell viability.The cells were divided into control group and 200μmol/L H_(2)O_(2) group,and were subjected to 0,2,4,6,8 Gy radiotherapy,respectively,and the clonogenic assay was performed to assess the radiosensitizing effect of H_(2)O_(2).The cells were divided into control group,200μmol/L H_(2)O_(2) group,4 Gy group and 200μmol/L H_(2)O_(2)+4 Gy group,and the expressions of autophagy-related proteins were measured by Western blotting assay and immunofluorescence assay.Results The expressions of autophagy-related proteins LC3 II and p62 between control group and sodium hyaluronate group was not statistically significant(P>0.05).The expressions of autophagy-related proteins LC3 II and p62 between 4 Gy group and sodium hyaluronate+4 Gy group was not statistically significant(P>0.05).The results of CCK8 experiment showed that the cell proliferation rate was not statistically significant between control group and sodium hyaluronate group before and after radiotherapy(P>0.05);compared with control group,200μmol/L H_(2)O_(2) promoted cell proliferation(P<0.05),500μmol/L H_(2)O_(2) had no statistically significant effect on cell proliferation(P>0.05),and 700μmol/L H_(2)O_(2) and 1000μmol/L H_(2)O_(2) inhibited cell proliferation(P<0.05).The results of clonogenesis assay showed that H_(2)O_(2) increased the sensitivity of cells to radiotherap
作者
任慧欣
郑茂金
韩文灿
王超群
周云
裴冬生
REN Huixin;ZHENG Maojin;HAN Wencan;WANG Chaoqun;ZHOU Yun;PEI Dongsheng(Laboratory of Clinical and Experimental Pathology Xuzhou Medical University,Xuzhou 221004,Jiangsu,China;Department of Radiotherapy,Xuzhou Clinical College,Xuzhou Medical University,Xuzhou 221004,Jiangsu,China)
出处
《山东大学学报（医学版）》
CAS
北大核心
2023年第6期22-28,共7页
Journal of Shandong University:Health Sciences
基金
2020年度徐州医科大学-面上项目(XYFM2020013)。
