microRNA-20a靶向调控唾液酸转移酶ST8SIA2在结直肠癌耐药中的作用  

Effect of microRNA-20a on the colorectal cancer drug resistance by targeting ST8SIA2

作　　者：宫晓红 王丹 沈宗坤 贾莉[4] 刘铃 GONG Xiao-hong;WANG Dan;SHEN Zong-kun;JIA Li;LIU Ling(Department of Clinical Laboratory,Dalian Traditional Chinese Medical Hospital,Liaoning 116013,China;Department of Clinical Laboratory,General Hospital of Anshan Iron and Steel Group Corporation,Liaoning 114002,China;Department of Clinical Laboratory,Fuxin Central Hospital,Liaoning 123000,China;College of Laboratory Medicine,Dalian Medical University,Liaoning 116044,China;Department of Clinical Laboratory,Dandong First Hospital,Liaoning 118000,China)

机构地区：[1]大连中医医院检验科,辽宁116013 [2]鞍钢集团公司总医院检验科,辽宁114002 [3]阜新中心医院检验科,辽宁123000 [4]大连医科大学检验医学院,辽宁116044 [5]丹东市第一医院检验科,辽宁118000

出　　处：《中国医药生物技术》2018年第6期514-519,共6页Chinese Medicinal Biotechnology

摘　　要：目的探究基于miR-20a为探针调控ST8SIA2在结直肠癌耐药中的功能和机制,为寻求逆转药物提供新的治疗策略及靶点。方法采用RT-PCR、Western blot分别检测结直肠癌Lovo细胞及其耐药细胞株Lovo/5-FU中miR-20a、ST8SIA2的表达水平;利用CCK-8及Western blot实验检测上调、下调mi R-20a后两种细胞株的药物敏感性及其ST8SIA2的表达情况;靶基因预测及双荧光素酶报告实验验证miR-20a与ST8SIA2的靶向关系;利用CCK-8实验进一步检测Lovo细胞中miR-20a和ST8SIA2表达调控对细胞药物敏感性的影响。结果①与Lovo细胞相比,miR-20a在耐药细胞株Lovo/5-FU中呈现高表达,ST8SIA2呈现低表达;②特异性上调Lovo细胞中miR-20a表达,ST8SIA2表达减弱,该细胞的药物敏感性减弱;③特异性下调Lovo/5-FU细胞中miR-20a表达,ST8SIA2表达增加,该细胞的药物敏感性增强;④特异性上调Lovo细胞中ST8SIA2表达,可逆转miR-20a对细胞产生的作用。结论 miR-20a及ST8SIA2的表达水平与结直肠癌耐药性密切相关,miR-20a可能是通过靶向负调控ST8SIA2的表达进而调控结直肠癌细胞的耐药。Objective We aim to explore the mechanism of miR-20a to regulate ST8SIA2 in colorectal cancer drug resistance and to provide a novel therapeutic strategy and target of colorectal cancer. Methods The expression of miR-20a and ST8SIA2 was analyzed by real-time PCR and Western blot in colorectal cancer cells. CCK-8 and Western blot assays were used to detect the drug sensitivity and the ST8SIA2 expression in the two cell lines after up-regulation and downregulation of miR-20a, respectively. The interaction between miR-20a and its predicted target gene ST8SIA2 was confirmed by 3＇-UTR dual-luciferase reporter assay. The cell viability was obtained to further investigate the drug sensitivity of Lovo cells upon transfection with miR-20a mimic and ST8SIA2. Results miR-20a was highly expressed and ST8SIA2 lowly expressed in Lovo/5-FU cells. Overexpression of miR-20a in Lovo cells could decrease ST8SIA2 level, and decrease the chemosensitivity to anti-tumor drugs. Silencing of miR-20a in Lovo/5-FU cells resulted in the increased expression of ST8SIA2, and increased the chemosensitivity to anti-tumor drugs. Overexpression of ST8SIA2 could reversed the effect of miR-20a mimic on drug resistance. Conclusion The levels of miR-20a and ST8SIA2 were closely related to the drug resistance in colorectal cancer. miR-20a could regulate drug resistance in colorectal cancer cells by targeting ST8SIA2 expression.

关 键 词：结直肠肿瘤 miR-20a ST8SIA2 抗药性 肿瘤 

分 类 号：R735.34[医药卫生—肿瘤]