长链非编码RNA uc002ktr.3在肺鳞癌顺铂耐药中的作用研究

The role of long non-coding RNA uc002ktr.3 on drug resistance to cisplatin in lung squamous cancer

目的研究肺鳞癌顺铂敏感和耐受组织中长链非编码RNA的表达差异,探讨长链非编码RNA uc002ktr.3在肺鳞癌顺铂耐药中的作用。方法利用ArraystarlncRNA芯片技术检测顺铂敏感和耐药的肺鳞癌组织中lncRNAs;通过siRNA干扰技术降低肺鳞癌SK-MES-1细胞中uc002ktr.3表达,CCK-8法检测细胞半数抑制浓度(IC 50 )的变化,Western blot法检测SK-MES-1细胞中Zeb1和E-cadherin蛋白变化,流式细胞仪检测细胞凋亡变化。结果聚类分析顺铂耐药和敏感的肺鳞癌组织中长链非编码RNA表达谱显示,共有 1 220 个lncRNAs存在差异性表达(差异倍数>2.0);CCK-8实验表明,干扰uc002ktr.3的SK-MES-1组相对于阴性对照(siRNA-NC)组的IC 50 显著降低;Western blot证实,干扰uc002ktr.3可显著降低SK-MES-1细胞中Zeb1蛋白表达而增加E-cadherin蛋白;凋亡检测结果显示,在相同剂量顺铂药物作用下,干扰uc002ktr.3后SK-MES-1细胞凋亡率显著高于siRNA-NC组。结论长链非编码RNA uc002ktr.3在肺鳞癌顺铂耐药组织中表达显著升高,干扰uc002ktr.3表达可明显增加肺鳞癌顺铂药物敏感性,uc002ktr.3可作为治疗肺鳞癌顺铂耐药的重要分子靶标。

Objective To investigate the differential expression of long non-coding RNA ( lncRNA) in cisplatin sensitive and resistant tissues of lung squamous cancer,and to explore the role of long non-coding RNA uc002ktr.3 in cisplatin resistance of lung squamous cancer. Methods LncRNAs in lung squamous carcinoma with cisplatin sensitivity and cisplatin resistance were detected by ArraystarlncRNA chip technology.SiRNA transfection was applied to knockdown uc002ktr.3 expression in SK-MES-1.The drug sensitivity (IC 50 ) of SK-MES-1 were detected by CCK-8 assay;the protein expression of Zeb1 and E-cadherin were detected by Western blot and apoptosis rates were detected by flow cytometry. Results A total of 1220 lncRNAs were differentially expressed in cisplatin resistant compared with cisplatin sensitive lung squamous cancer (Fold change>2.0).The CCK-8 assay and Western blot results showed that knockdown of uc002ktr.3 in SK-MES-1 could significantly reduce the drug sensitivity (IC 50 ) and the expression of Zeb1 protein,however,the expression of E-cadherin protein was dramatically increased,comparing with negative control (siRNA-NC) group.Flow cytometry analysis showed that,treatment with the same dose of cisplain, knockdown of uc002ktr.3 in SK-MES-1 could significantly increased apoptosis rates compared with negative control (siRNA-NC) group. Conclusion Long non-coding RNA uc002ktr.3 is significantly up-regulated in lung squamous cancer with cisplatin resistance.Our findings indicate that the decrease of uc002ktr.3 can significantly improve the cell susceptibility to cisplatin.These findings provide new vision and implications for targeted therapy of lung squamous cancer.