Long non-coding RNA GAS5 inhibits DDP-resistance and tumor progression of epithelial ovarian cancer via GAS5-E2F4-PARP1-MAPK axis

Abstract 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 ovar...

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Main Authors: Xiaoran Long, Keqi Song, Hao Hu, Qi Tian, Wenjing Wang, Qian Dong, Xia Yin, Wen Di
Format: Article
Language:English
Published: BMC 2019-08-01
Series:Journal of Experimental & Clinical Cancer Research
Subjects:
Online Access:http://link.springer.com/article/10.1186/s13046-019-1329-2
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spelling doaj-64dca5d90b4940f8829bf00eb1e815482020-11-25T02:59:16ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662019-08-0138111610.1186/s13046-019-1329-2Long non-coding RNA GAS5 inhibits DDP-resistance and tumor progression of epithelial ovarian cancer via GAS5-E2F4-PARP1-MAPK axisXiaoran Long0Keqi Song1Hao Hu2Qi Tian3Wenjing Wang4Qian Dong5Xia Yin6Wen Di7Department of Obstetrics and Gynecology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong UniversityDepartment of Obstetrics and Gynecology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong UniversityDepartment of Cancer Intervention, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong UniversityDepartment of Obstetrics and Gynecology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong UniversityDepartment of Obstetrics and Gynecology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong UniversityDepartment of Obstetrics and Gynecology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong UniversityDepartment of Obstetrics and Gynecology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong UniversityDepartment of Obstetrics and Gynecology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong UniversityAbstract 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 νs. 3 normal ovary tissues) and RT- qPCR (53 EOC tissues νs. 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.http://link.springer.com/article/10.1186/s13046-019-1329-2lncRNA GAS5PARP1E2F4MAPKOvarian cancer
collection DOAJ
language English
format Article
sources DOAJ
author Xiaoran Long
Keqi Song
Hao Hu
Qi Tian
Wenjing Wang
Qian Dong
Xia Yin
Wen Di
spellingShingle Xiaoran Long
Keqi Song
Hao Hu
Qi Tian
Wenjing Wang
Qian Dong
Xia Yin
Wen Di
Long non-coding RNA GAS5 inhibits DDP-resistance and tumor progression of epithelial ovarian cancer via GAS5-E2F4-PARP1-MAPK axis
Journal of Experimental & Clinical Cancer Research
lncRNA GAS5
PARP1
E2F4
MAPK
Ovarian cancer
author_facet Xiaoran Long
Keqi Song
Hao Hu
Qi Tian
Wenjing Wang
Qian Dong
Xia Yin
Wen Di
author_sort Xiaoran Long
title Long non-coding RNA GAS5 inhibits DDP-resistance and tumor progression of epithelial ovarian cancer via GAS5-E2F4-PARP1-MAPK axis
title_short Long non-coding RNA GAS5 inhibits DDP-resistance and tumor progression of epithelial ovarian cancer via GAS5-E2F4-PARP1-MAPK axis
title_full Long non-coding RNA GAS5 inhibits DDP-resistance and tumor progression of epithelial ovarian cancer via GAS5-E2F4-PARP1-MAPK axis
title_fullStr Long non-coding RNA GAS5 inhibits DDP-resistance and tumor progression of epithelial ovarian cancer via GAS5-E2F4-PARP1-MAPK axis
title_full_unstemmed Long non-coding RNA GAS5 inhibits DDP-resistance and tumor progression of epithelial ovarian cancer via GAS5-E2F4-PARP1-MAPK axis
title_sort long non-coding rna gas5 inhibits ddp-resistance and tumor progression of epithelial ovarian cancer via gas5-e2f4-parp1-mapk axis
publisher BMC
series Journal of Experimental & Clinical Cancer Research
issn 1756-9966
publishDate 2019-08-01
description Abstract 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 νs. 3 normal ovary tissues) and RT- qPCR (53 EOC tissues νs. 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.
topic lncRNA GAS5
PARP1
E2F4
MAPK
Ovarian cancer
url http://link.springer.com/article/10.1186/s13046-019-1329-2
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