摘要
目的验证长链非编码RNA SOX2OT(Long noncoding RNA SOX2OT,SOX2OT)增强大肠癌HCT-116细胞多药耐药(Multidrug resistance,MDR),并从SOX2OT/miR-34a/SOX2信号通路探讨其可能的有效机制。方法双荧光素酶报告基因验证SOX2OT与miR-34a、miR-34a与SOX2的靶向结合;HCT-116细胞瞬时转染SOX2OT质粒、miR-34a mimic,RT-qPCR验证转染效率;RT-qPCR检测SOX2OT、miR-34a和SOX2表达水平并进行相关性分析;CCK-8检测细胞增殖,评估SOX2OT、miR-34a对HCT-116细胞5-氟尿嘧啶(5-FU)、长春新碱(VCR)、顺铂(CDDP)、紫杉醇(TAXOL)化疗敏感性的影响。结果SOX2OT与miR-34a、miR-34a与SOX2之间存在靶向关系;SOX2OT抑制miR-34a(P<0.05),上调SOX2(P<0.01);miR-34下调SOX2OT(P<0.01)和SOX2(P<0.01);转染SOX2OT质粒后HCT-116细胞的5-FU、VCR、CDDP、TAXOL的IC50值均升高(P<0.01);HCT-116细胞共转染SOX2OT、miR-34a mimic时对四种化疗药物的IC50值与转染SOX2OT组相比均明显降低(P<0.01),其中5-FU、CDDP的IC50值与对照组相比无统计学差异(P>0.05)。结论SOX2OT海绵吸附miR-34a促进SOX2的表达致使HCT-116细胞多药耐药。
Objective The aim of this study was to verify that long noncoding RNA SOX2OT(SOX2OT)enhanced the multidrug resistance(MDR)of colorectal cancer HCT-116 cells,and explored its possible effective mechanism on SOX2OT/miR-34a/SOX2 signaling pathway.Methods Dual luciferase reporter gene assay was used to verify the targeted binding of SOX2OT to miR-34a,and miR-34a to SOX2;The SOX2OT plasmid and miR-34a were transient transfected into HCT-116 cells,and the transfection efficiency was verified by RT-qPCR;RT-qPCR was used to detect the levels of SOX2OT,miR-34a and SOX2,and the correlation amongst them were analyzed;The CCK-8 assay was used to detect cell proliferation and to assess the influence of SOX2OT and miR-34a on the chemosensitivity of HCT-116 cells to 5-fluorouracil(5-FU),vincristine(VCR),cisplatin(CDDP),and paclitaxel(Taxol).Results There was a targeting relationship between SOX2OT and miR-34a,and miR-34a with SOX2;The SOX2OT inhibited miR-34a(P<0.05)and upregulated the level of SOX2(P<0.01);The MiR-34a down-regulated the expression of SOX2OT and SOX2(P<0.01);The IC50 values for 5-FU,VCR,CDDP,and TAXOL in HCT-116 cells were increased after transfection of SOX2OT plasmid(P<0.01);The IC50 values of four chemotherapy drugs in the co-transfected with SOX2OT and miR-34a mimic groups were significantly lower than those of the transfected SOX2OT group(P<0.01).The IC50 values of 5-FU and CDDP were not different from the control group(P>0.05).Conclusion The SOX2OT sponge adsorbs miR-34a and promotes the expression of SOX2,resulting in multidrug resistance of HCT-116 cells.
作者
郭飘婷
邹阳
GUO Piaoting;ZOU Yang(Department of General Medicine,The Second Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine,Hangzhou 310005,China;Department of Orthopedics,The Second Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine)
出处
《实用肿瘤学杂志》
CAS
2020年第5期398-403,共6页
Practical Oncology Journal
基金
国家自然科学基金青年项目(编号:81803876)。
