MiR-574-5p promotes cell proliferation by negatively regulating small C-terminal domain phosphatase 1 in esophageal squamous cell carcinoma

Document Type : Original Article


1 Department of Human Anatomy, Xuzhou Medical University, Xuzhou, Jiangsu, China

2 Jiangsu Medical Engineering Research Center of Gene Detection, Xuzhou Medical University, Xuzhou, Jiangsu, China

3 Department of Forensic Medicine, Xuzhou Medical University, Xuzhou, Jiangsu, China

4 Department of Basic Pathology, Pathology College, Qiqihar Medical University, Qiqihar, Heilongjiang, China

5 NGS center, Hangzhou D.A. Medical Laboratory Co., Ltd., Hangzhou, Zhejiang, China

6 Department of Orthopedics, the Affiliated Hospital of Xuzhou Medical University, Xuzhou, China


Objective(s): Esophageal cancer is one of the most common cancers with high incidence and mortality rates, especially in China. MicroRNA (miRNA) can be used as a prognostic marker for various human cancers. This study aims to detect suitable miRNA markers for esophageal squamous cell carcinoma (ESCC). 
Materials and Methods: Our previous gene expression data of ESCC cells and the data from GSE43732 and GSE112840 were analyzed. The expression of miR-574-5p in ESCC patients and controls was analyzed by real-time quantitative PCR. The effect of miR-574-5p on proliferation was detected by real-time cell analysis (RTCA) and EdU proliferation assay after cell transfections. The target gene small C-terminal domain phosphatase 1 (CTDSP1) of miR-574-5p was validated by luciferase reporter assay and western blotting.
Results: In the current study, the bioinformatics analysis found miR-574-5p up-regulated in ESCC. The qPCR assay of 26 ESCC and 13 adjacent/ normal tissues confirmed these results. We further demonstrated that miR-574-5p overexpression promoted cell proliferation. Then the dual-luciferase reporter assay and the rescue experiment suggested that CTDSP1 was a direct target of miR-574-5p.
Conclusion: MiR-574-5p played an oncological role in ESCC by interacting and negatively regulating CTDSP1. These results provided a deeper understanding of the effect of miR-574-5p on ESCC.


1.    Murphy G, McCormack V, Abedi-Ardekani B, Arnold M, Camargo MC, Dar NA, et al. International cancer seminars: A focus on esophageal squamous cell carcinoma. Ann Oncol 2017; 28: 2086-2093.
2.    Arnold M, Soerjomataram I, Ferlay J, Forman D. Global incidence of oesophageal cancer by histological subtype in 2012. Gut 2015; 64: 381-387.
3.    Nayak B, Khan N, Garg H, Rustagi Y, Singh P, Seth A, et al. Role of miRNA-182 and miRNA-187 as potential biomarkers in prostate cancer and its correlation with the staging of prostate cancer. Int Braz J Urol 2020; 46: 614-623.
4.    Ali AE, Abd ES, Mohamed MA, Swellam M. Clinical role of MiRNA 29a and MiRNA 335 on breast cancer management: their relevance to MMP2 protein level. Arch Physiol Biochem 2022;128:1058-1065.
5.    Elghoroury EA, ElDine HG, Kamel SA, Abdelrahman AH, Mohammed A, Kamel MM, et al. Evaluation of miRNA-21 and miRNA Let-7 as Prognostic Markers in Patients With Breast Cancer. Clin Breast Cancer 2018; 18: e721-e726.
6.    Li F, Wang F, Zhu C, Wei Q, Zhang T, Zhou YL. miR-221 suppression through nanoparticle-based miRNA delivery system for hepatocellular carcinoma therapy and its diagnosis as a potential biomarker. Int J Nanomedicine 2018; 13: 2295-2307.
7.    Cui J, Qi S, Liao R, Su D, Wang Y, Xue S. MiR-574-5p promotes the differentiation of human cardiac fibroblasts via regulating ARID3A. Biochem Biophys Res Commun 2020; 521: 427-433.
8.    He B, Zhou W, Rui Y, Liu L, Chen B, Su X. MicroRNA-574-5p attenuates acute respiratory distress syndrome by targeting HMGB1. Am J Respir Cell Mol Biol 2021; 64: 196-207.
9.    Zhou X, Liang Z, Qin S, Ruan X, Jiang H. Serum-derived miR-574-5p-containing exosomes contribute to liver fibrosis by activating hepatic stellate cells. Mol Biol Rep 2022; 49: 1945-1954.
10.    Hegewald AB, Breitwieser K, Ottinger SM, Mobarrez F, Korotkova M, Rethi B, et al. Extracellular miR-574-5p induces osteoclast differentiation via TLR 7/8 in rheumatoid arthritis. Front Immunol 2020, 11: 585282.
11.    Donzelli J, Proestler E, Riedel A, Nevermann S, Hertel B, Guenther A, et al. Small extracellular vesicle-derived miR-574-5p regulates PGE2-biosynthesis via TLR7/8 in lung cancer. J Extracell Vesicles 2021; 10: e12143.
12.    Sun Y, Yi Y, Gan S, Ye R, Huang C, Li M, et al. miR-574-5p mediates epithelial-mesenchymal transition in small cell lung cancer by targeting vimentin via a competitive endogenous RNA network. Oncol lett 2021; 21: 459.
13.    Saul MJ, Baumann I, Bruno A, Emmerich AC, Wellstein J, Ottinger SM, et al. miR-574-5p as RNA decoy for CUGBP1 stimulates human lung tumor growth by mPGES-1  induction. FASEB J 2019; 33: 6933-6947.
14.    Lin Z, Chen M, Wan Y, Lei L, Ruan H. miR-574-5p targets FOXN3 to regulate the invasion of nasopharyngeal carcinoma cells via Wnt/beta-catenin pathway. Technol Cancer Res Treat 2020; 19: 1079239307.
15.    Zhang ZJ, Xiao Q, Li XY. MicroRNA-574-5p directly targets FOXN3 to mediate thyroid cancer progression via  Wnt/beta-catenin signaling pathway. Pathol Res Pract 2020, 216: 152939.
16.    Zhang S, Zhang R, Xu R, Shang J, He H, Yang Q. MicroRNA-574-5p in gastric cancer cells promotes angiogenesis by targeting protein tyrosine phosphatase non-receptor type 3 (PTPN3). Gene 2020; 733: 144383.
17.    Tong R, Zhang J, Wang C, Li Q, Wang L, Ju M. Inhibition of miR-574-5p suppresses cell growth and metastasis and enhances chemosensitivity by targeting RNA binding protein QKI in cervical cancer cells. Naunyn Schmiedebergs Arch Pharmacol 2020, 393: 951-966.
18.    Zhang Z, Li X, Xiao Q, Wang Z. MiR-574-5p mediates the cell cycle and apoptosis in thyroid cancer cells via Wnt/beta-catenin signaling by repressing the expression of Quaking proteins. Oncol Lett 2018; 15: 5841-5848.
19.    Han GL, Wang J, Guo K, Chen JK, Shang RX, Jiang T. miRNA-574-5p downregulates ZNF70 and influences the progression of human esophageal squamous cell carcinoma through reactive oxygen species generation and MAPK pathway activation. Anticancer Drugs 2020; 31: 282-291.
20.    Yang B, Kuai F, Chen Z, Fu D, Liu J, Wu Y, et al. miR-634 decreases the radioresistance of human breast cancer cells by targeting STAT3. Cancer Biother Radiopharm 2020; 35: 241-248.
21.    Shan G, Tang T, Xia Y, Qian HJ. MEG3 interacted with miR-494 to repress bladder cancer progression through targeting PTEN. J Cell Physiol 2020; 235: 1120-1128.
22.    Cui Z, Liu G, Kong D. miRNA27a promotes the proliferation and inhibits apoptosis of human pancreatic cancer cells by Wnt/beta-catenin pathway. Oncol Rep 2018; 39: 755-763.
23.    Rallabandi HR, Ganesan P, Kim YJ. Targeting the C-Terminal Domain Small Phosphatase 1. Life (Basel) 2020; 10:57.
24.    Krasnov GS, Puzanov GA, Afanasyeva MA, Dashinimaev EB, Vishnyakova KS, Beniaminov AD, et al. Tumor suppressor properties of the small C-terminal domain phosphatases in non-small cell lung cancer. Biosci Rep 2019; 39: BSR20193094.
25.    Gu Z, Eils R, Schlesner M. Complex heatmaps reveal patterns and correlations in multidimensional genomic data. Bioinformatics 2016; 32: 2847-2849.
26.    Chen Z, Li J, Tian L, Zhou C, Gao Y, Zhou F, et al. MiRNA expression profile reveals a prognostic signature for esophageal squamous cell carcinoma. Cancer Lett 2014; 350: 34-42.
27.    Zheng D, Ding Y, Ma Q, Zhao L, Guo X, Shen Y, et al. Identification of Serum MicroRNAs as Novel Biomarkers in Esophageal Squamous Cell Carcinoma Using Feature Selection Algorithms. Front oncol 2018, 8: 674.
28.    Su H, Hu N, Yang HH, Wang C, Takikita M, Wang QH, et al. Global gene expression profiling and validation in esophageal squamous cell carcinoma and its association with clinical phenotypes. Clin cancer res 2011; 17: 2955-2966.
29.    Agarwal V, Bell GW, Nam JW, Bartel DP. Predicting effective microRNA target sites in mammalian mRNAs. Elife 2015; 4: e05005.
30.    Lebourgeois S, Fraisse A, Hennechart-Collette C, Guillier L, Perelle S, Martin-Latil S. Development of a real-time cell analysis (RTCA) method as a fast and accurate method for detecting infectious particles of the adapted strain of hepatitis a virus. Front Cell Infect Microbiol 2018; 8: 335.
31.    Liao P, Wang W, Li Y, Wang R, Jin J, Pang W, et al. Palmitoylated SCP1 is targeted to the plasma membrane and negatively regulates angiogenesis. Elife 2017; 6: e22058.
32.    Yang M, Liu R, Sheng J, Liao J, Wang Y, Pan E, et al. Differential expression profiles of microRNAs as potential biomarkers for the early diagnosis of esophageal squamous cell carcinoma. Oncol Rep 2013; 29: 169-176.
33.    Li J, Han X, Gu Y, Wu J, Song J, Shi Z, et al. LncRNA MTX2-6 Suppresses cell proliferation by acting as ceRNA of miR-574-5p to accumulate SMAD4 in esophageal squamous cell carcinoma. Front Cell Dev Biol 2021, 9: 654746.
34.    Kohn M. Turn and Face the Strange: A new view on phosphatases. ACS Cent Sci 2020; 6: 467-477.
35.    Rasko JE, Wong JJ. Nuclear microRNAs in normal hemopoiesis and cancer. J Hematol Oncol 2017; 10: 8.
36.    Emmerich AC, Wellstein J, Ossipova E, Baumann I, Lengqvist J, Kultima K, et al. Proteomics-based characterization of miR-574-5p decoy to CUGBP1 suggests specificity for mPGES-1 regulation in human lung cancer cells. Front pharmacol 2020; 11: 196.
37.    Qian W, Li Q, Wu X, Li W, Li Q, Zhang J, et al. Deubiquitinase USP29 promotes gastric cancer cell migration by cooperating with phosphatase SCP1 to stabilize Snail protein. Oncogene 2020; 39: 6802-6815.
38.    Ma LM, Liang ZR, Zhou KR, Zhou H, Qu LH. 27-Hydroxycholesterol increases Myc protein stability via suppressing PP2A, SCP1  and FBW7 transcription in MCF-7 breast cancer cells. Biochem Biophys Res Commun 2016; 480: 328-333.
39.    Sun AG, Wang MG, Li B, Meng FG. Down-regulation of miR-124 target protein SCP-1 inhibits neuroglioma cell migration. Eur Rev Med Pharmacol Sci 2017; 21: 723-729.
40.    Gervasi NM, Dimtchev A, Clark DM, Dingle M, Pisarchik AV, Nesti LJ. C-terminal domain small phosphatase 1 (CTDSP1) regulates growth factor expression and axonal regeneration in peripheral nerve tissue. Sci Rep 2021; 11: 14462.
41.    Wang W, Liao P, Shen M, Chen T, Chen Y, Li Y, et al. SCP1 regulates c-Myc stability and functions through dephosphorylating c-Myc Ser62. Oncogene 2016, 35: 491-500.
42.    Sun T, Fu J, Shen T, Lin X, Liao L, Feng XH, et al. The Small C-terminal domain phosphatase 1 inhibits cancer cell migration and invasion by dephosphorylating Ser(P)68-twist1 to accelerate twist1 protein degradation. J Biol Chem 2016, 291: 11518-11528.
43.    Revathidevi S, Munirajan AK. Akt in cancer: Mediator and more. Semin cancer Biol 2019; 59: 80-91.