miR-210表达对肝癌细胞SMMC-7721放疗的影响  

Effect of miR-210 expression on radiotherapy of liver cancer cell line SMMC-7721

作　　者：李顺延 高飞[2] 郭虎林 LI Shun-yan;GAO Fei;GUO Hu-lin(The Fifth People′s Hospital of Qinghai Province,Xining,Qinghai 810007,China;The Second Affiliated Hospital of Dalian Medical University,Dalian,Liaoning 116031,China)

机构地区：[1]青海省第五人民医院,810007 [2]大连医科大学附属第二医院,116031

出　　处：《现代消化及介入诊疗》2020年第9期1180-1184,共5页Modern Interventional Diagnosis and Treatment in Gastroenterology

基　　金：大连市医学科学研究计划项目(1712033)。

摘　　要：目的观察miR-210对肝癌细胞SMMC-7721放疗敏感性的影响,并探讨其对肝癌细胞SMMC-7721肿瘤的作用机制。方法采用脂质体Lipofectamine TM 2000转染的方法,将化学合成的miR-210mimics和miR-210 inhibitor瞬时转染肝癌细胞系SMMC-7721,分别构建稳定过表达miR-210细胞株组(miR-210 mimic组)、稳定沉默miR-210表达细胞株组(miR-210 inhibitor组),阴性对照组(miR-210 NC组)及空白对照组。RT-PCR检测各组细胞中miR-210的相对表达水平;克隆存活实验检测细胞放射敏感性变化。将四组人肝癌SMMC-7721细胞分别接种于裸鼠皮下。待各组裸鼠肿瘤直径达到6~8 mm,构建SMMC-7721细胞株裸鼠移植瘤模型,24 h后各组裸鼠肿瘤局部接受8 Gy照射,测量各组裸鼠放疗后不同时间的肿瘤体积,记算各组裸鼠的肿瘤生长延缓时间(TGD),辐射增敏系数和平均存活时间;照后24 h用免疫组化染色检测肿瘤间质微血管密度。结果与空白对照组和miR-210 NC组比较,miR-210 mimics组miR-210相对表达量升高(P<0.05),miR-210相对表达量降低(P<0.05)。下调miR-210表达能增强SMMC-7721细胞放射敏感性。miR-210 mimic组肿瘤达到6mm需要的时间显著少于miR-210 NC组及空白对照组(P<0.05),miR-210 inhibitor组显著多于miR-210 NC组及空白对照组(P<0.05);肿瘤8 Gy照射后,miR-210 inhibitor组肿瘤平均生存时间显著高于miR-210 NC组及空白对照组(P<0.05),miR-210 mimic组显著低于miR-210 NC组及空白对照组(P<0.05);miR-210 inhibitor组肿瘤体积抑制率、生长延缓时间均高于miR-210 NC组及空白对照组,miR-210 mimic组均低于miR-210 NC组及空白对照组;照射后24 h,miR-210 mimic组的肿瘤间质微血管密度大于miR-210 NC组及空白对照组(P<0.05),miR-210 inhibitor组的肿瘤间质微血管密度小于miR-210 NC组及空白对照组(P<0.05)。结论低表达miR-210能增强人肝癌SMMC-7721细胞放射敏感性,够延迟裸鼠移植入肝癌细胞成瘤时间,抑制移植瘤增殖和血管生成,Objective To observe the effect of mir-210 on the radiotherapy sensitivity of hepatocellular carcinoma cell smmc-7721,and to explore its mechanism of action on hepatocellular carcinoma cell smmc-7721 tumor.Methods Mir-210mimics and mir-210 inhibitor were transfected into hepatocellular carcinoma cell line smmc-7721 with LipofectamineTM2000,and stable mir-210 expressed cell line(mir-210 inhibitor),negative control group(mir-210-nc)and blank control groupwere constructed,respectivelyRt-pcr was used to detect the relative expression level of mir-210 in each group.Clone survival experiments to detect cellular radiosensitivity changes will four groups of human liver cancer SMMC-7721 cells were inoculated subcutaneously in nude mice with groups of nude mice tumor diameter to 6 to 8 mm,SMMC-7721 cell lines nude mouse transplantation tumor model was constructed,after 24 h groups of nude mice tumor local exposure to 8 Gy,measured at different times after radiotherapy of tumor volume,groups of nude mice of account groups of nude mice tumor growth delay time(TGD),radiation sensitization coefficient and the average survival time;Tumor interstitial microvascular density was detected by immunohistochemical staining 24 h after irradiation.Results Compared with blank control group and mir-210 NC group,the relative expression of mir-210 in mir-210 mimics group was increased(t=2.334,3.167;P<0.05),and the relative expression of mir-210 was decreased(t=2.188,2.397;P<0.05).Down regulation of mir-210 expression can enhance the radiosensitivity of SMMC-7721 cells.The time required for the tumor to reach 6mm in the miR-210 mimic group was significantly less than that in the miR-210-NC group and the blank control group,while that in the miR-210 inhibitor group was significantly higher than that in the miR-210-NC group and the blank control group(tactile 3.289 and 3.454).After 8 Gy irradiation,the average survival time of tumor in miR-210 inhibitor group was significantly higher than that in miR-210-NC group and blank control group,while

关 键 词：MicroRNA-210 肝癌细胞SMMC-7721 放疗 

分 类 号：R614[医药卫生—麻醉学] R735.7[医药卫生—外科学]