Long non-coding RNA GAS5 inhibits DDP-resistance and tumor progression of epithelial ovarian cancer via GAS5-E2F4-PARP1-MAPK axis
Long non-coding RNA GAS5 inhibits DDP-resistance and tumor progression of epithelial ovarian cancer via GAS5-E2F4-PARP1-MAPK axis
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长链非编码RNA GAS5通过GAS5-E2F4-PARP1-MAPK轴抑制上皮性卵巢癌的顺铂耐药及肿瘤进展
DOI：
10.1186/s13046-019-1329-2
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发表时间：
2019-08-07
影响因子：
11.3
通讯作者：
Di, Wen
中科院分区：
文献类型：
--
作者：
Long, Xiaoran;Song, Keqi;Di, Wen
研究背景上皮性卵巢癌（Epithelial ovarian cancer，EOC）是女性生殖系统中病死率最高的恶性肿瘤。卵巢上皮性癌早期对顺铂等化疗药物的耐受性是其预后不良的重要因素之一。在这里，我们的目的是研究一个特定的长非编码RNA，lncRNA GAS 5的失调，及其在EOC进展中的作用。方法采用基因芯片和实时定量PCR技术检测卵巢癌组织和卵巢癌细胞株中lncRNAGAS 5的低表达。流式细胞仪检测OC细胞的细胞周期和凋亡。CCK-8法检测OC细胞对顺铂的敏感性。Western blot检测OC细胞系中细胞生长标志物、凋亡标志物、PARP 1、E2 F4、MAPK通路蛋白表达及其他蛋白表达。RNA pull-down和RIP实验证明GAS 5和E2 F4结合。通过CHIP-qPCR检测和荧光素酶报告基因检测来确定E2 F4对PARP 1的影响。在体外和体内评估lncRNA GAS 5对OC细胞的作用。结果通过芯片（3例EOC组织nu. 3例正常卵巢组织）和RT-qPCR（53例卵巢上皮癌组织nu. 10例正常卵巢组织）中，我们鉴定出lncRNA GAS 5在EOC样品中显著低表达并与预后相关。与敏感细胞株相比，GAS 5在顺铂耐药OC细胞中也呈低表达，GAS 5的过表达可显著增强OC细胞对顺铂的体内外敏感性。同时GAS 5的过表达也导致OC细胞G 0/G1期阻滞和凋亡增加。GAS 5可能通过募集转录因子E2 F4到其启动子来调节PARP 1的表达，进而影响MAPK通路的活性。由于GAS 5的5 'TOP结构，其在OC细胞中可被转录抑制剂雷帕霉素调控。结论本研究探讨了lncRNA-GAS 5导致卵巢癌顺铂耐药和肿瘤进展的具体机制，首次提出了GAS 5-E2 F4-PARP 1-MAPK轴及其在卵巢癌药物敏感性和进展中的作用，为临床应用提供了实验依据。
Background Epithelial ovarian cancer (EOC) is the malignant tumor of the female reproductive system with the highest fatality rate. Tolerance of chemotherapeutic drugs like cisplatin (DDP) occurring in very early stage is one of the important factors of the poor prognosis of epithelial ovarian cancer. Here we aim to study the dysregulation of a particular long noncoding RNA, lncRNA GAS5, and its role in EOC progression. Methods The low expression of lncRNA GAS5 in EOC tissues and OC cell lines was determined by microarray analyses and Real-Time qPCR. Flow cytometer assays were used to detect cell cycle and apoptosis of OC cells. CCK8 assay were performed to investigate the DDP sensitivity of OC cells. Western blot was carried out to detect cell growth markers, apoptotic markers, PARP1, E2F4, MAPK pathway protein expression and other protein expression in OC cell lines. The binding of GAS5 and E2F4 were proved by RNA pull-down and RIP assay. The effect of E2F4 on PARP1 were determined by CHIP-qPCR assay and luciferase reporter assay. The effect of lncRNA GAS5 on OC cells was assessed in vitro and in vivo. Results By microarray (3 EOC tissues nu s. 3 normal ovary tissues) and RT- qPCR (53 EOC tissues nu s. 10 normal ovary tissues) we identified lncRNA GAS5 to be dramatically low expressed in EOC samples and correlated with prognosis. Compared with sensitive cell lines, GAS5 was also low expressed in DDP resistant OC cell lines, and over-expression of GAS5 significantly enhanced the sensitivity of OC cells to DDP in vivo and in vitro. Meanwhile the over-expression of GAS5 also caused OC cells G0/G1 arrest and apoptosis increase. Mechanistically, GAS5 might regulate PARP1 expression by recruiting the transcription factor E2F4 to its promoter, and then affect the MAPK pathway activity. Due to the 5'TOP structure, GAS5 could be regulated by transcription inhibitor rapamycin in OC cells. Conclusion Here we explored the specific mechanisms of EOC cisplatin resistance and tumor progress due to lncRNA-GAS5, presented the GAS5-E2F4-PARP1-MAPK axis and its role in OC drug-sensitivity and progression for the first time, and the results may provide experimental basis for clinical application.