微小RNA-155促进SKOV3、A2780和原代培养的卵巢癌细胞凋亡。
MicroRNA-155 promotes apoptosis in SKOV3, A2780, and primary cultured ovarian cancer cells.
作者信息
Chen Wei, Huang Liuxuan, Hao Chenjun, Zeng Wenshu, Luo Xu, Li Xiaodi, Zhou Longshu, Jiang Songshan, Chen Zheng, He Yuanli
机构信息
Department of Obstetrics and Gynecology, Zhujiang Hospital, Southern Medical University, Guangzhou, 510260, China.
Department of Gynecology, The Second Affiliated Hospital of Guangzhou Medical University, 250 East Changgang Road, Guangzhou, 510260, China.
出版信息
Tumour Biol. 2016 Jul;37(7):9289-99. doi: 10.1007/s13277-016-4804-9. Epub 2016 Jan 15.
MicroRNAs (miRNAs) are a large group of small non-coding RNAs that can negatively regulate gene expression at the post-transcriptional level. The deregulation of miRNAs has been associated with tumorigenesis, drug resistance, and prognosis in cancers. Deregulated miR-155 has been reported in numerous cancers; however, its function remains unclear. 4',6-Diamidino-2-phenylindole (DAPI) staining and terminal-deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) techniques were used to determine the effects of a miR-155 mimic or inhibitor on the apoptotic ratio of ovarian cancer cells induced by cisplatin. Bioinformatic predictions, the dual-luciferase reporter assay, and western blot analysis were used to detect how miR-155 regulates X-linked inhibitor of apoptosis protein (XIAP). We demonstrated that a miR-155 mimic could decrease the IC50 value of cisplatin in SKOV3 ovarian cancer cells. Subsequently, gain- and loss-of-function analyses with a miR-155 mimic and inhibitor showed that miR-155 sensitizes ovarian cancer cells to cisplatin. Furthermore, the results from the luciferase assays and western blot analysis identified XIAP as the direct target of miR-155. In addition, introducing XIAP cDNA without a three prime untranslated region (3'-UTR) rescued the miR-155 promotion of apoptosis. These results indicate that miR-155 mediates cisplatin-induced apoptosis by targeting XIAP in ovarian cancer cells and that miR-155 could be a potential therapeutic target to increase the efficiency of ovarian cancer interventions.
微小RNA(miRNA)是一大类小的非编码RNA,可在转录后水平负向调节基因表达。miRNA的失调与肿瘤发生、耐药性及癌症预后相关。在众多癌症中均报道有miR-155失调;然而,其功能仍不清楚。采用4',6-二脒基-2-苯基吲哚(DAPI)染色和末端脱氧核苷酸转移酶介导的缺口末端标记(TUNEL)技术来确定miR-155模拟物或抑制剂对顺铂诱导的卵巢癌细胞凋亡率的影响。运用生物信息学预测、双荧光素酶报告基因检测及蛋白质免疫印迹分析来检测miR-155如何调节X连锁凋亡抑制蛋白(XIAP)。我们证明miR-155模拟物可降低顺铂在SKOV3卵巢癌细胞中的半数抑制浓度(IC50)值。随后,用miR-155模拟物和抑制剂进行的功能获得和功能缺失分析表明,miR-155使卵巢癌细胞对顺铂敏感。此外,荧光素酶检测和蛋白质免疫印迹分析结果确定XIAP是miR-155的直接靶标。另外,导入无3'非翻译区(3'-UTR)的XIAP cDNA可挽救miR-155对凋亡的促进作用。这些结果表明,miR-155通过靶向卵巢癌细胞中的XIAP介导顺铂诱导的凋亡,且miR-155可能是提高卵巢癌干预效率的潜在治疗靶点。