长链非编码RNA SNHG6通过在结肠癌细胞中吸附miR-26a-5p，经ULK1诱导的自噬促进化疗耐药。
LncRNA SNHG6 promotes chemoresistance through ULK1-induced autophagy by sponging miR-26a-5p in colorectal cancer cells.
作者信息
Wang Xinke, Lan Zhixian, He Juan, Lai Qiuhua, Yao Xiang, Li Qingyuan, Liu Yongfeng, Lai Huasheng, Gu Chuncai, Yan Qun, Fang Yuxin, Zhang Yue, Li Aimin, Liu Side
机构信息
Guangdong Provincial Key Laboratory of Gastroenterology, Department of Gastroenterology, Nanfang Hospital, Southern Medical University, No. 1838, Guangzhou Avenue North, Guangzhou, People's Republic of China.
出版信息
Cancer Cell Int. 2019 Sep 9;19:234. doi: 10.1186/s12935-019-0951-6. eCollection 2019.
BACKGROUND
Chemotherapy resistance is one of the main causes of recurrence in colorectal cancer (CRC) patients and leads to poor prognosis. Long noncoding RNAs (lncRNAs) have been reported to regulate chemoresistance. We aimed to determine the role of the lncRNA small nucleolar RNA host gene 6 (SNHG6) in CRC cell chemoresistance.
METHODS
Cell drug sensitivity tests and flow cytometry were performed to analyze CRC cell chemoresistance. Animal models were used to determine chemoresistance in vivo, and micro RNA (miRNA) binding sites were detected by dual-luciferase reporter assays. Bioinformatics analysis was performed to predict miRNAs binding to SNHG6 and target genes of miR-26a-5p. SNHG6/miR-26a-5p/ULK1 axis and autophagy-related proteins were detected by qRT-PCR and western blotting. Furthermore, immunofluorescence was employed to confirm the presence of autophagosomes.
RESULTS
SNHG6 enhanced CRC cell resistance to 5-fluorouracil (5-FU), promoted autophagy, inhibited 5-FU-induced apoptosis, and increased 5-FU resistance in vivo. Bioinformatics analysis showed that miR-26a-5p might bind to SNHG6 and target ULK1, and dual-luciferase reporter assays confirmed this activity. qRT-PCR and western blotting showed that SNHG6 was able to negatively regulate miR-26a-5p but correlated positively with ULK1.
CONCLUSION
SNHG6 may promote chemoresistance through ULK1-induced autophagy by sponging miR-26a-5p in CRC cells.
背景
化疗耐药是结直肠癌（CRC）患者复发的主要原因之一，且会导致预后不良。据报道，长链非编码RNA（lncRNA）可调节化疗耐药性。我们旨在确定长链非编码RNA小核仁RNA宿主基因6（SNHG6）在CRC细胞化疗耐药中的作用。
方法
进行细胞药物敏感性试验和流式细胞术以分析CRC细胞的化疗耐药性。使用动物模型确定体内化疗耐药性，并通过双荧光素酶报告基因检测法检测微小RNA（miRNA）结合位点。进行生物信息学分析以预测与SNHG6结合的miRNA以及miR-26a-5p的靶基因。通过qRT-PCR和蛋白质免疫印迹法检测SNHG6/miR-26a-5p/ULK1轴和自噬相关蛋白。此外，采用免疫荧光法确认自噬体的存在。
结果
SNHG6增强了CRC细胞对5-氟尿嘧啶（5-FU）的耐药性，促进了自噬，抑制了5-FU诱导的细胞凋亡，并增加了体内对5-FU的耐药性。生物信息学分析表明，miR-26a-5p可能与SNHG6结合并靶向ULK1，双荧光素酶报告基因检测法证实了这一活性。qRT-PCR和蛋白质免疫印迹法显示，SNHG6能够负向调节miR-26a-5p，但与ULK1呈正相关。
结论
在CRC细胞中，SNHG6可能通过海绵吸附miR-26a-5p，经由ULK1诱导的自噬促进化疗耐药。
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