Effectsof laryngeal cancer exosomal miR-20a in regulating tumor-associated macrophage proliferation and apoptosis
DU Li1, YUAN Yaozong2, WANG Hui3, and GAO Hongsheng4
1. Department of Laboratory Pathology, 2. Department of Anesthesiology , Xinjiang Production and Construction Corps Hospital of Chinese People’s Armed Police Force, Urumqi 830063, China; 3. Otolaryngology Head and Neck Surgery, General Hospital of Xinjiang Military Region, Urumqi 830000, China; 4. Logistics University of Chinese People’s Armed Police Force, Tianjin 300162, China
Abstract：Objective To explore the effectsof laryngeal carcinoma exosomal miR-20a on the proliferation and apoptosis of tumor-associated macrophages. Methods The exosomes secreted by 16HBE, TU686, TU177 and TU212 cells were extracted and identified (recorded as 16HBE exo, TU686 exo, TU177 exo and TU212 exo). TU212 cells were transfected with miR-NC and miR-20a mimic, and the exosomes secreted by TU212after transfection were collected (recorded as miR-NC exo and miR-20a exo). M2 macrophages were incubated with PBS, 16HBE exo, TU686 exo, TU177 exo, TU212 exo, miR-NC exoand miR-20a exo for 48 h (listed as PBS group, 16HBE exo group, TU686 exo group, TU177 exo group, TU212 exo group, miR-NC exo group and miR-20a exo group). The proliferation ability of cells was detected by CCK8 method, the apoptosis rate of cells was detected by flow cytometry,and PI3K and AKT phosphorylation levels in cells in each group were detected by Western blotting. Results The extracted exosomes were consistent with their structural and biological characteristics. The proliferation ability of M2 type macrophages in the TU686 exo group, TU177 exo group, and TU212 exo group were significantly higher than that in the PBS group (P<0.05), the apoptosis rate (9.67%±1.53%, 8.67%±2.08%, 4.00%±1.00%) was significantly lower than that in the PBS group (21.67%±4.72%) (P<0.01), the phosphorylation levels of PI3K (0.81±0.03, 0.70±0.03, 1.10±0.08) and AKT (1.55±0.11, 1.86±0.06, 2.13±0.10) were significantly higher than those in the PBS group (0.19±0.04 and 0.11±0.03) (P<0.001), the proliferation ability of M2 type macrophages in the miR-20a exo group was significantly higher than that in the miR-NC exo group (P<0.05), and the apoptosis rate (8.33%±1.52%) was significantly lower than that in the miR-NC exo group (14.33%±1.53%) (P<0.01). The phosphorylation levels of PI3K (1.05±0.09) and AKT (1.55±0.11) were significantly higher than those in the miR-NC exo group (0.11±0.04 and 1.19±0.03) (P<0.001). Conclusions Laryngeal carcinoma exosomal miR-20a can regulate the proliferation and apoptosis of M2 macrophages, and the mechanism may be the activation of PI3K/AKT signaling pathway.
Zhang L, Yu D. Exosomes in cancer development, metastasis, and immunity [J]. Biochim Biophys Acta Rev Cancer, 2019, 1871(2): 455-468.
[5]
He C, Li L, Wang L, et al. Exosome-mediated cellular crosstalk within the tumor microenvironment upon irradiation[J]. Cancer Biol Med, 2021, 18(1):21-33.
[6]
Han C, Zhang C, Wang H, et al. Exosome-mediated communication between tumor cells and tumor-associated macrophages: implications for tumor microenvironment[J]. Oncoimmunology, 2021, 10(1):1887552-1887567.
Yue B, Yang H, Wang J, et al. Exosome biogenesis, secretion and function of exosomal miRNAs in skeletal muscle myogenesis[J]. Cell Prolif, 2020, 53(7): e12857-e12866.
[9]
Shi L, Zhu W, Huang Y, et al. Cancer-associated fibroblast-derived exosomal microRNA-20a suppresses the PTEN/PI3K-AKT pathway to promote the progression and chemoresistance of non-small cell lung cancer[J]. Clin Transl Med, 2022, 12(7):e989-e1012.
[10]
Moloudizargari M, Hekmatirad S, Mofarahe Z S, et al. Exosomal microRNA panels as biomarkers for hematological malignancies[J]. Curr Probl Cancer, 2021, 45(5):100726-100734.
[11]
Oishi Y, Manabe I. Macrophages in inflammation, repair and regeneration[J]. Int Immunol, 2018, 30(11):511-528.
Li B, Cao Y, Sun M, et al. Expression, regulation, and function of exosome-derived miRNAs in cancer progression and therapy[J]. FASEB J, 2021, 35(10):e21916-e21927.
[14]
Fan Z, Yu J, Lin J, et al. Exosome-specific tumor diagnosis via biomedical analysis of exosome-containing microRNA biomarkers [J]. Analyst, 2019, 144(19):5856-5865.
[15]
Yu F, Lin Y, Tan G, et al. Tumor-derived exosomal microRNA-15b-5p augments laryngeal cancer by targeting TXNIP[J]. Cell Cycle, 2022, 21(7):730-740.
[16]
Zhao Q, Zheng X, Guo H, et al. Serum Exosomal miR-941 as a promising Oncogenic Biomarker for Laryngeal Squamous Cell Carcinoma[J]. J Cancer, 2020, 11(18):5329-5344.
[17]
Xu Z, Zhao L, Zhu L Y, et al. MicroRNA-17, 20a regulates the proangiogenic function of tumor-associated macrophages via targeting hypoxia-inducible factor 2α[J]. PLoS One, 2013, 8(10):e77890-e77898.
[18]
Petty A J, Owen DH, Yang Y, et al. Targeting Tumor-Associated Macrophages in Cancer Immunotherapy[J]. Cancers, 2021, 13(21):5318-5336.
[19]
Lopez-Yrigoyen M, Cassetta L, Pollard J W. Macrophage targeting in cancer[J]. Ann N Y Acad Sci, 2021, 1499(1):18-41.
[20]
Mantovani A, Marchesi F, Malesci A, et al. Tumour-associated macrophages as treatment targets in oncology[J]. Nat Rev Clin Oncol, 2017, 14(7):399-416.
[21]
Su C, Jia S, Ma Z, et al. HMGB1 promotes lymphangiogenesis through the activation of RAGE on M2 macrophages in laryngeal squamous cell carcinoma [J]. Dis Markers, 2022, 4(20):4487435-4487452.
[22]
Guo Y, Feng Y, Cui X, et al. Autophagy inhibition induces the repolarisation of tumour-associated macrophages and enhances chemosensitivity of laryngeal cancer cells to cisplatin in mice [J]. Cancer Immunol Immunother, 2019, 68(12):1909-1920.
[23]
Zhao S, Mi Y, Guan B, et al. Tumor-derived exosomal miR-934 induces macrophage M2 polarization to promote liver metastasis of colorectal cancer[J]. J Hematol Oncol, 2020, 19, 13(1):156-174.
[24]
Lin F, Yin H B, Li X Y, et al. Bladder cancer cell-secreted exosomal miR-21 activates the PI3K/AKT pathway in macrophages to promote cancer progression[J]. Int J Oncol, 2020, 56(1):151-164.
[25]
He Y, Sun M M, Zhang G G, et al. Targeting PI3K/Akt signal transduction for cancer therapy[J]. Signal Transduct Target Ther, 2021, 6(1): 425-441.
2023, Vol. 34 
