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Journal of ZheJiang University(Medical Science)  2016, Vol. 45 Issue (4): 364-370    DOI: 10.3785/j.issn.1008-9292.2016.07.06
    
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
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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 wordsEsophageal 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     
Received: 12 October 2015     
CLC:  R73-3  
  R735.1  
Cite this article:

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.

URL:

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


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

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 
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