TGF-β1 modulates temozolomide resistance in glioblastoma via altered microRNA processing and elevated MGMT

被引：58
作者：
Nie, Er [1 ]
Jin, Xin [2 ]
Miao, Faan [1 ]
Yu, Tianfu [3 ]
Zhi, Tongle [4 ]
Shi, Zhumei [5 ]
Wang, Yingyi [5 ]
Zhang, Junxia [5 ]
Xie, Manyi [1 ]
You, Yongping [5 ]
机构：
[1] Xuzhou Med Univ, Affiliated Hosp, Dept Neurosurg, 99 Huaihai West Rd, Xuzhou 221000, Jiangsu, Peoples R China
[2] Nanjing Gaochun Peoples Hosp, Dept Med, Nanjing, Jiangsu, Peoples R China
[3] Nanjing Univ, Affiliated Drum Tower Hosp, Sch Med, Dept Neurosurg, Nanjing, Jiangsu, Peoples R China
[4] Nantong Univ, Affiliated Hosp 4, Yancheng City Peoples Hosp 1, Dept Neurosurg, Yancheng, Jiangsu, Peoples R China
[5] Nanjing Med Univ, Affiliated Hosp 1, Dept Neurosurg, 300 Guangzhou Rd, Nanjing 210029, Jiangsu, Peoples R China
来源：
NEURO-ONCOLOGY | 2021年 / 23卷 / 03期
基金：
中国国家自然科学基金;
关键词：
glioblastoma; KSRP; MGMT; temozolomide resistance; TGF-beta; 1; RNA-BINDING PROTEIN; GROWTH-FACTOR-BETA; TGF-BETA; KSRP; PROMOTES; METHYLATION;
D O I：
10.1093/neuonc/noaa198
中图分类号：
R73 [肿瘤学];
学科分类号：
100214 ;
摘要：
Background. Our previous studies have indicated that miR-198 reduces cellular methylguanine DNA methyltransferase (MGMT) levels to enhance temozolomide sensitivity. Transforming growth factor beta 1 (TGF-beta 1) switches off miR-198 expression by repressing K-homology splicing regulatory protein (KSRP) expression in epidermal keratinocytes. However, the underlying role of TGF-beta 1 in temozolomide resistance has remained unknown. Methods. The distribution of KSRP was detected by western blotting and immunofluorescence. Microarray analysis was used to compare the levels of long noncoding RNAs (lncRNAs) between TGF-beta 1-treated and untreated cells. RNA immunoprecipitation was performed to verify the relationship between RNAs and KSRP. Flow cytometry and orthotopic and subcutaneous xenograft tumor models were used to determine the function of TGF-beta 1 in temozolomide resistance. Results. Overexpression of TGF-beta 1 contributed to temozolomide resistance in MGMT promoter hypomethylated glioblastoma cells in vitro and in vivo. TGF-beta 1 treatment reduced cellular MGMT levels through suppressing the expression of miR-198. However, TGF-beta 1 upregulation did not affect KSRP expression in glioma cells. We identified and characterized 2 lncRNAs (H19 and HOXD-AS2) that were upregulated by TGF-beta 1 through Smad signaling. H19 and HOXD-AS2 exhibited competitive binding to KSRP and prevented KSRP from binding to primary miR-198, thus decreasing miR-198 expression. HOXD-AS2 or H19 upregulation strongly promoted temozolomide resistance and MGMT expression. Moreover, KSRP depletion abrogated the effects of TGF-beta 1 and lncRNAs on miR-198 and MGMT. Finally, we found that patients with low levels of TGF-beta 1 or lncRNA expression benefited from temozolomide therapy. Conclusions. Our results reveal an underlying mechanism by which TGF-beta 1 confers temozolomide resistance. Furthermore, our findings suggest that a novel combination of temozolomide with a TGF-beta inhibitor may serve as an effective therapy for glioblastomas.
引用
收藏
页码：435 / 446
页数：12
相关论文
共 32 条
[1]   Predicting protein associations with long noncoding RNAs [J].
Bellucci, Matteo ;
Agostini, Federico ;
Masin, Marianela ;
Tartaglia, Gian Gaetano .
NATURE METHODS, 2011, 8 (06) :444-445
[2]   Functional and molecular insights into KSRP function in mRNA decay [J].
Briata, Paola ;
Chen, Ching-Yi ;
Ramos, Andres ;
Gherzi, Roberto .
BIOCHIMICA ET BIOPHYSICA ACTA-GENE REGULATORY MECHANISMS, 2013, 1829 (6-7) :689-694
[3]   High TGFβ-Smad activity confers poor prognosis in glioma patients and promotes cell proliferation depending on the methylation of the PDGF-B gene [J].
Bruna, Alejandra ;
Darken, Rachel S. ;
Rojo, Federico ;
Ocana, Alberto ;
Penuelas, Silvia ;
Arias, Alexandra ;
Paris, Raquel ;
Tortosa, Avelina ;
Mora, Jaume ;
Baselga, Jose ;
Seoane, Joan .
CANCER CELL, 2007, 11 (02) :147-160
[4]   Coordinated Regulation of Polycomb Group Complexes through microRNAs in Cancer [J].
Cao, Qi ;
Mani, Ram-Shankar ;
Ateeq, Bushra ;
Dhanasekaran, Saravana M. ;
Asangani, Irfan A. ;
Prensner, John R. ;
Kim, Jung H. ;
Brenner, J. Chad ;
Jing, Xiaojun ;
Cao, Xuhong ;
Wang, Rui ;
Li, Yong ;
Dahiya, Arun ;
Wang, Lei ;
Pandhi, Mithil ;
Lonigro, Robert J. ;
Wu, Yi-Mi ;
Tomlins, Scott A. ;
Palanisamy, Nallasivam ;
Qin, Zhaohui ;
Yu, Jindan ;
Maher, Christopher A. ;
Varambally, Sooryanarayana ;
Chinnaiyan, Arul M. .
CANCER CELL, 2011, 20 (02) :187-199
[5]   The genesis and evolution of homeobox gene clusters [J].
Garcia-Fernàndez, J .
NATURE REVIEWS GENETICS, 2005, 6 (12) :881-892
[6]   The sequence selectivity of KSRP explains its flexibility in the recognition of the RNA targets [J].
Garcia-Mayoral, Maria Flor ;
Diaz-Moreno, Irene ;
Hollingworth, David ;
Ramos, Andres .
NUCLEIC ACIDS RESEARCH, 2008, 36 (16) :5290-5296
[7]   The structure of the C-terminal KH domains of KSRP reveals a noncanonical motif important for mRNA degradation [J].
Garcia-Mayoral, Maria Flor ;
Hollingworth, David ;
Masino, Laura ;
Diaz-Moreno, Irene ;
Kelly, Geoff ;
Gherzi, Roberto ;
Chou, Chu-Fan ;
Chen, Ching-Yi ;
Ramos, Andres .
STRUCTURE, 2007, 15 (04) :485-498
[8]   A KH domain RNA binding protein, KSRP, promotes ARE-directed mRNA turnover by recruiting the degradation machinery [J].
Gherzi, R ;
Lee, KY ;
Briata, P ;
Wegmüller, D ;
Moroni, C ;
Karin, M ;
Chen, CY .
MOLECULAR CELL, 2004, 14 (05) :571-583
[9]   H19 long noncoding RNA controls the mRNA decay promoting function of KSRP [J].
Giovarelli, Matteo ;
Bucci, Gabriele ;
Ramos, Andres ;
Bordo, Domenico ;
Wilusz, Carol J. ;
Chen, Ching-Yi ;
Puppo, Margherita ;
Briata, Paola ;
Gherzi, Roberto .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2014, 111 (47) :E5023-E5028
[10]  
Han JF, 2015, AM J CANCER RES, V5, P945
1234