Identification of FBXW7α-regulated genes in M1-polarized macrophages in colorectal cancer by RNA sequencing
Objectives: To determine the FBXW7α-regulated genes in tumor-polarized macrophages in colorectal cancer.
Methods: This experimental study was performed between June 2017 and March 2019. FBXW7α siRNA transfected RAW264.7 cells, together with the control group, were co-cultured with the colon cancer cell line, Colon-26. M1 marker production from the macrophages was determined by ELISA and quantitative reverse transcription-polymerase chain reaction. Whole genomic differential expression between the FBXW7α siRNA group and the control group were determined by RNA-sequencing analysis. The target site of the microRNA-205 gene was predicted using Targetscan and was verified by the luciferase assay. By transfecting mimics or inhibitors of microRNA-205, we explored the role of FBXW7α/microRNA-205 axis in regulating the polarization of tumor-associated macrophages (TAM).
Results: FBXW7α knockdown in RAW264.7 enhanced the expression of cyclooxigenase (COX)-2 and inducible nitric oxide synthase (iNOS), mRNA expression and IL6, IL12, p40, and tumor necrosis factor-α (TNFα) production upon co-culture with Colon-26 cells in vitro. Further, compared with the control group, 648 genes in total were enhanced and 416 targets were downregulated in FBXW7α siRNA transfected cells, among which miR-205 was the most significantly upregulated. SMAD1 was identified as an miR-205 target. The FBXW7α/miR-205 axis might regulate TAM polarization by affecting SMAD1 expression.
Conclusion: These results prove that the FBXW7α/miR-205 axis plays an important role in TAM polarization and could facilitate further exploration of its molecular mechanism.
Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal
A. Global cancer statistics, 2012. CA Cancer J Clin 2015; 65:
Isidro RA, Appleyard CB. Colonic macrophage polarization
in homeostasis, inflammation, and cancer. Am J Physiol
Gastrointest Liver Physiol 2016; 311: G59-G73.
Peng L, Zhang H, Hao Y, Xu F, Yang J, Zhang R, et al.
Reprogramming macrophage orientation by microRNA 146b
targeting transcription factor IRF5. EBioMedicine 2016; 14:
Yeh CH, Bellon M, Nicot C. FBXW7: a critical tumor
suppressor of human cancers. Mol Cancer 2018; 17: 115.
Kothari N, Teer JK, Abbott AM, Srikumar T, Zhang Y, Yoder SJ,
et al. Increased incidence of FBXW7 and POLE proofreading
domain mutations in young adult colorectal cancers. Cancer
; 122: 2828-2835.
Xie T, Cho YB, Wang K, Huang D, Hong HK, Choi YL, et al.
Patterns of somatic alterations between matched primary and
metastatic colorectal tumors characterized by whole-genome
sequencing. Genomics 2014; 104: 234-241.
Mlecnik B, Bindea G, Kirilovsky A, Angell HK, Obenauf
AC, Tosolini M, et al. The tumor microenvironment and
Immunoscore are critical determinants of dissemination to
distant metastasis. Sci Transl Med 2016; 8: 327ra26.
Zeng Y, Zhu J, Shen D, Qin H, Lei Z, Li W, et al. Repression
of Smad4 by miR-205 moderates TGF-β-induced epithelialmesenchymal
transition in A549 cell lines. Int J Oncol 2016;
Saxena A, Fayad R, Kaur K, Truman S, Greer J, Carson JA, et al.
Dietary selenium protects adiponectin knockout mice against
chronic inflammation induced colon cancer. Cancer Biol Ther
; 18: 257-267.
O’Malley G, Heijltjes M, Houston AM, Rani S, Ritter T, Egan
LJ, et al. Mesenchymal stromal cells (MSCs) and colorectal
cancer: a troublesome twosome for the anti-tumour immune
response? Oncotarget 2016; 7: 60752-60774.
Mantovani A, Marchesi F, Malesci A, Laghi L, Allavena P.
Tumour-associated macrophages as treatment targets in
oncology. Nat Rev Clin Oncol 2017; 14: 399-416.
Petty AJ, Yang Y. Tumor-associated macrophages: implications
in cancer immunotherapy. Immunotherapy 2017; 9: 289-302.
Wang H, Chen B, Duan B, Zheng J, Wu X. miR‑205 suppresses
cell proliferation, invasion, and metastasis via regulation of the
PTEN/AKT pathway in renal cell carcinoma. Mol Med Rep
; 14: 3343-3349.
Ji T, Zhang X, Li W. microRNA‑205 acts as a tumor suppressor
and directly targets YAP1 in glioma. Mol Med Rep 2017; 16:
Means AL, Freeman TJ, Zhu J, Woodbury LG, Marincola-
Smith P, Wu C, et al. Epithelial smad4 deletion up-regulates
inflammation and promotes inflammation-associated cancer.
Cell Mol Gastroenterol Hepatol 2018; 6: 257-276.
Shi L, Xi J, Xu X, Peng B, Zhang B. MiR-148a suppressed cell
invasion and migration via targeting WNT10b and modulating
β-catenin signaling in cisplatin-resistant colorectal cancer cells.
Biomed Pharmacother 2019; 109: 902-909.
Flamini V, Jiang WG, Cui Y. Therapeutic role of MiR-140-5p
for the treatment of non-small cell lung cancer. Anticancer Res
; 37: 4319-4327.
Melman YF, Shah R, Danielson K, Xiao J, Simonson B, Barth
A, et al. Circulating microRNA-30d is associated with response
to cardiac resynchronization therapy in heart failure and
regulates cardiomyocyte apoptosis: a translational pilot study.
Circulation 2015; 131: 2202-2216.
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