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浙江大学学报（医学版）
浙江大学学报（医学版）  2016, Vol. 45 Issue (4): 364-370    DOI: 10.3785/j.issn.1008-9292.2016.07.06
癌分子医学专题     
靶向RAD18的小干扰RNA对人食管鳞癌ECA-109细胞增殖和化疗敏感性的影响
娄鹏荣1,2, 孙晓南1, 周俊东3, 邹士涛4
1. 浙江大学医学院附属邵逸夫医院放疗科, 浙江 杭州 310020;
2. 浙江省宁波市第一医院肿瘤放疗科, 浙江 宁波 315010;
3. 南京医科大学附属苏州医院放疗科, 江苏 苏州 215001;
4. 南京医科大学附属苏州医院苏州肿瘤中心重点实验室, 江苏 苏州 215001
Effect of RAD18-siRNA on proliferation and chemotherapy sensitivity of human esophageal squamous cell carcinoma ECA-109 cells
LOU Pengrong1,2, SUN Xiaonan1, ZHOU Jundong3, ZOU Shitao4
1. Department of Radio-Oncology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310020, China;
2. Department of Radio-Oncology, Ningbo First Hospital, Ningbo 315010, China;
3. Department of Radio-Oncology, Suzhou Municipal Hospital, Nanjing Medical University, Suzhou 215001, China;
4. Suzhou Cancer Center Core Laboratory, Suzhou Municipal Hospital, Nanjing Medical University, Suzhou 215001, China
全文: PDF(1060 KB)  
摘要: 

目的：研究靶向RAD18基因的小干扰RNA（siRNA）对食管鳞癌细胞ECA-109增殖和化疗敏感性的影响。方法：合成针对RAD18基因的siRNA（RAD18-siRNA），通过脂质体将其转染人食管鳞癌ECA-109细胞株，采用荧光定量PCR方法检测ECA-109 RAD18和CyclinD1 mRNA的表达，蛋白质印迹法检测其RAD18和CyclinD1蛋白的表达；应用CCK-8法检测ECA-109细胞的增殖和化疗药物对ECA-109细胞存活率的影响，流式细胞术检测ECA-109细胞周期；采用Pearson检验分析RAD18与CyclinD1基因表达的相关性。结果：与未转染组比较，RAD18-siRNA组RAD18表达明显降低（P<0.05）。RAD18-siRNA组细胞增殖抑制（P<0.05），G1期细胞数增多，G2/M期细胞数减少（P<0.05）。不同浓度顺铂或5-氟尿嘧啶处理细胞后，两组细胞的存活率均下降（均P<0.05），但RAD18-siRNA组细胞半数抑制浓度较未转染组减少（P<0.05）。在食管鳞癌组织中，RAD18基因mRNA的表达与CyclinD1基因mRNA的表达呈正相关（r=0.478，P<0.01）。结论：下调RAD18表达能够抑制食管鳞癌细胞的增殖，提高其对化疗药物的敏感性；CyclinD1在食管鳞癌中的表达水平可能参与这一过程。
关键词 ： 食管肿瘤/药物疗法,  癌,鳞状细胞/药物疗法,  RNA,小分子干扰/治疗应用,  基因,肿瘤抑制/治疗应用,  细胞增殖/药物作用,  细胞周期蛋白D1/药物作用,  细胞系,肿瘤    
Abstract：

Objective: To investigate the effect of RAD18-siRNA on cell proliferation and chemotherapy sensitivity of esophageal squamous cell carcinoma (ESCC) ECA-109 cells. Methods: RAD18-siRNA was transfected into human ECA-109 cells by Lipofectamine 3000. Quantitative PCR and Western blot were performed to detect RAD18 and CyclinD1 expression; CCK-8 assay was used to determine cell proliferation and chemotherapy drug sensitivity; flow cytometry was used to determine cell cycle. Correlation between RAD18 and CyclinD1 mRNA expression was analyzed by Pearson's correlation. Results: Compared with non-transfected cells, the expression of RAD18 in RAD18-siRNA group was significantly decreased (P<0.05). The cell proliferation was inhibited (P<0.05) and the cell number of G1 phase was increased, G2/M phase cells decreased (P<0.05) in RAD18-siRNA group. After treatment with different concentrations of cisplatin or 5-FU, the survival rate of the two cell groups was reduced (all P<0.05), and the IC50 of RAD18-siRNA group was significantly lower than that of non-transfected group (P<0.05). The mRNA expression of RAD18 was positively correlated with CyclinD1 expression in ESCC tissues(r=0.478, P<0.01). Conclusion: Down-regulated expression of RAD18 can decrease the cell proliferation and increase chemo-sensitivity of ESCC cells, and CyclinD1 may participate in the process.
Key words： Esophageal neoplasms/drug therapy    Carcinoma, squamous cell/drug therapy    RNA, small interfering/therapeutic use    Genes, tumor suppressor/therapeutic use    Cell proliferation/drug effects    CyclinD1/drug effects    Cell line, tumor
收稿日期: 2015-10-12
CLC:  R73-3  
基金资助:

国家自然科学基金（81372433）
通讯作者: 孙晓南(1966-),E-mail:sunxiaonan@hotmail.com;周俊东(1976-),E-mail:zhoujundong330@163.com     E-mail: sunxiaonan@hotmail.com;zhoujundong330@163.com
作者简介: 娄鹏荣(1978-),男,硕士研究生,副主任医师,从事肿瘤放化疗研究;E-mail:68046039@qq.com;http://orcid.org/0000-0001-7349-5145
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引用本文:

娄鹏荣 等. 靶向RAD18的小干扰RNA对人食管鳞癌ECA-109细胞增殖和化疗敏感性的影响[J]. 浙江大学学报（医学版）, 2016, 45(4): 364-370.
LOU Pengrong, SUN Xiaonan, ZHOU Jundong, ZOU Shitao. Effect of RAD18-siRNA on proliferation and chemotherapy sensitivity of human esophageal squamous cell carcinoma ECA-109 cells. Journal of ZheJiang University(Medical Science), 2016, 45(4): 364-370.

链接本文:

http://www.zjujournals.com/xueshu/med/CN/10.3785/j.issn.1008-9292.2016.07.06      或      http://www.zjujournals.com/xueshu/med/CN/Y2016/V45/I4/364

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