TLE3 represses colorectal cancer proliferation by inhibiting MAPK and AKT signaling pathways.
BACKGROUND
Transducin-like enhancer of Split3 (TLE3) serves as a transcriptional corepressor during cell differentiation and shows multiple roles in different kinds of cancers. Recently, TLE3 together with many other genes involved in Wnt/β-catenin pathway were detected hyper-methylated in colorectal cancer (CRC). However, the potential role and the underlying mechanism of TLE3 in CRC progression remain scarce.


METHODS
Gene expression profiles were analyzed in The Cancer Genome Atlas (TCGA) microarray dataset of 41 normal colorectal intestine tissues and 465 CRC tissues. Western blot and Real-time Quantitative PCR (RT-qPCR) were respectively performed to detect protein and mRNA expression in 8 pairs of CRC tissue and matched adjacent normal mucosa. Immunohistochemistry (IHC) was conducted to evaluate TLE3 protein expression in 105 paraffin-embedded, archived human CRC tissues from patients, whose survival data were analyzed with Kaplan-Meier method. In vitro experiments including MTT assay, colony formation assay, and soft agar formation assay were used to investigate the effects of TLE3 on CRC cell growth and proliferation. Additionally, subcutaneous tumorigenesis assay was performed in nude mice to confirm the effects of TLE3 in vivo. Furthermore, gene set enrichment analysis (GSEA) was run to explore potential mechanism of TLE3 in CRC, and then we measured the distribution of CRC cell cycle phases and apoptosis by flow cytometry, as well as the impacts of TLE3 on MAPK and AKT signaling pathways by Western blot and RT-qPCR.


RESULTS
TLE3 was significantly down-regulated in 465 CRC tissues compared with 41 normal tissues. Both protein and mRNA expressions of TLE3 were down-regulated in CRC compared with matched adjacent normal mucosa. Lower expression of TLE3 was significantly associated with poorer survival of patients with CRC. Besides, knock down of TLE3 promoted CRC cell growth and proliferation, while overexpression of TLE3 showed suppressive effects. Furthermore, overexpression of TLE3 caused G1-S phase transition arrest, inhibition of MAPK and AKT pathways, and up-regulation of p21Cip1/WAF1 and p27Kip1.


CONCLUSIONS
This study indicated that TLE3 repressed CRC proliferation partly through inhibition of MAPK and AKT signaling pathways, suggesting the possibility of TLE3 as a biomarker for CRC prognosis.
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Run-Wei
RW
Yang
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Ying-Yue
YY
Zeng
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Wen-Ting
WT
Wei
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Yan-Mei
YM
Cui
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Hui-Ying
HY
Sun
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Yue-Long
YL
Cai
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Xin-Xin
XX
Nian
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Yun-Teng
YT
Hu
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Yu-Ping
YP
Quan
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Sheng-Lu
SL
Jiang
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Meng
M
Wang
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Ya-Li
YL
Zhao
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Jun-Feng
JF
Qiu
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Ming-Xuan
MX
Li
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Jia-Huan
JH
Zhang
Guangdong Provincial Key Laboratory of Gastroenterology, Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, China.
Mei-Rong
MR
He
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China.
Li
L
Liang
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China. dyq@fimmu.com.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China. dyq@fimmu.com.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China. dyq@fimmu.com.|Department of Pathology, Nanfang Hospital and School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, Guangdong, China. dyq@fimmu.com.
Yan-Qing
YQ
Ding
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, Guangdong, China. liaowt2002@gmail.com.|Department of Pathology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong, China. liaowt2002@gmail.com.|Guangdong Provincial Key Laboratory of Molecular Tumor Pathology, Guangzhou, Guangdong, China. liaowt2002@gmail.com.|Department of Pathology, Nanfang Hospital and School of Basic Medical Sciences, Southern Medical University, Guangzhou, 510515, Guangdong, China. liaowt2002@gmail.com.
Wen-Ting
WT
Liao
Colorectal cancer; Prognosis; Proliferation; TLE3; p21Cip1/WAF1; p27Kip1
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