Fibroblast‑derived exosomal microRNA‑369 potentiates migration and invasion of lung squamous cell carcinoma cells via NF1‑mediated MAPK signaling pathway

Guo,Liping; Li,Baoli; Yang,Jianjun; Shen,Juan; Ji,Jinshan; Miao,Meijing

doi:10.3892/ijmm.2020.4614

Fibroblast‑derived exosomal microRNA‑369 potentiates migration and invasion of lung squamous cell carcinoma cells via NF1‑mediated MAPK signaling pathway

Authors:
- Liping Guo
- Baoli Li
- Jianjun Yang
- Juan Shen
- Jinshan Ji
- Meijing Miao
View Affiliations

Affiliations: Department of Pathology, Medical College of Yan’an University, Yan’an, Shaanxi 716000, P.R. China, Department of Pharmacology, Medical College of Yan'an University, Yan’an, Shaanxi 716000, P.R. China, Department of Interventional Radiology, Affiliated Hospital of Yan'an University, Yan’an, Shaanxi 716000, P.R. China, Department of Imaging, Medical College of Yan’an University, Yan’an, Shaanxi 716000, P.R. China, Department of Preventive Medicine, Medical College of Yan’an University, Yan’an, Shaanxi 716000, P.R. China, Department of Nursing, Medical College of Yan'an University, Yan'an, Shaanxi 716000, P.R. China
Published online on: May 22, 2020 https://doi.org/10.3892/ijmm.2020.4614
Pages: 595-608
Copyright: © Guo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Cancer‑associated fibroblasts (CAFs) exhibit tumor‑stimulating properties and are associated with poor survival in several types of cancer, making them potential therapeutic targets. The present study aimed to determine whether CAFs were associated with cell migration and invasion in lung squamous cell carcinoma (LUSC), as well as their association with microRNA‑369 (miR‑369) in these processes. Firstly, the changes of the malignant biological behavior were observed by treating the LUSC cells with the CAFs‑derived extracellular vesicles (CAFs‑EVs). Subsequently, the differentially expressed miRNAs in the cells treated with CAFs‑EVs were analyzed by microarray analysis. Following inhibition of miR‑369 expression in CAFs‑EVs, LUSC cells were co‑cultured, and the malignant biological behavior of the cells was re‑examined. Then, through bioinformatics analysis and verification, the mRNA targets of miR‑369 and the corresponding downstream signaling pathway were screened out. Finally, the effects of CAFs‑EVs on the growth and metastasis of LUSC were demonstrated by in vivo tumor formation and metastasis experiments. It was identified that miR‑369 was expressed at a relatively high level in the CAFs‑EVs. Neurofibromin‑1 (NF1) was hypothesized as a direct target of miR‑369 in LUSC. Also, the overexpression of miR‑369 activated the mitogen‑activated protein kinase signaling pathway by interacting with NF1, consequently potentiating LUSC cell growth. The present study provided novel insights into the action of miR‑369 in CAFs‑EVs in controlling LUSC cell migration, invasion and tumorigenesis, and identified miR‑369 in CAFs‑EVs as an important prognostic marker and therapeutic target.

View Figures

View References

1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2020. CA Cancer J Clin. 70:7–30. 2020. View Article : Google Scholar : PubMed/NCBI
2	Torre LA, Siegel RL and Jemal A: Lung Cancer Statistics. Adv Exp Med Biol. 893:1–19. 2016. View Article : Google Scholar
3	Xu F, Zhang H, Chen J, Lin L and Chen Y: Immune signature of T follicular helper cells predicts clinical prognostic and therapeutic impact in lung squamous cell carcinoma. Int Immunopharmacol. 81:1059322020. View Article : Google Scholar
4	Shi Y, Li Y, Yan C, Su H and Ying K: Identification of key genes and evaluation of clinical outcomes in lung squamous cell carcinoma using integrated bioinformatics analysis. Oncol Lett. 18:5859–5870. 2019.PubMed/NCBI
5	Erez N, Truitt M, Olson P, Arron ST and Hanahan D: Cancer-associated fibroblasts are activated in incipient neoplasia to orchestrate tumor-promoting inflammation in an NF-kappaB-dependent manner. Cancer Cell. 17:135–147. 2010. View Article : Google Scholar : PubMed/NCBI
6	Hanley CJ, Mellone M, Ford K, Thirdborough SM, Mellows T, Frampton SJ, Smith DM, Harden E, Szyndralewiez C, Bullock M, et al: Targeting the Myofibroblastic Cancer-Associated Fibroblast Phenotype Through Inhibition of NOX4. J Natl Cancer Inst. 110:2018. View Article : Google Scholar :
7	Raposo G and Stoorvogel W: Extracellular vesicles: Exosomes, microvesicles, and friends. J Cell Biol. 200:373–383. 2013. View Article : Google Scholar : PubMed/NCBI
8	Yang X, Li Y, Zou L and Zhu Z: Role of exosomes in crosstalk between cancer-associated fibroblasts and cancer cells. Front Oncol. 9:3562019. View Article : Google Scholar : PubMed/NCBI
9	Sun LP, Xu K, Cui J, Yuan DY, Zou B, Li J, Liu JL, Li KY, Meng Z and Zhang B: Cancer-associated fibroblast-derived exosomal miR3825p promotes the migration and invasion of oral squamous cell carcinoma. Oncol Rep. 42:1319–1328. 2019.PubMed/NCBI
10	Hao GJ, Ding YH, Wen H, Li XF, Zhang W, Su HY, Liu DM and Xie NL: Attenuation of deregulated miR-369-3p expression sensitizes non-small cell lung cancer cells to cisplatin via modulation of the nucleotide sugar transporter SLC35F5. Biochem Biophys Res Commun. 488:501–508. 2017. View Article : Google Scholar : PubMed/NCBI
11	Li JH, Liu S, Zhou H, Qu LH and Yang JH: starBase v2.0: Decoding miRNA-ceRNA, miRNA-ncRNA and protein-RNA interaction networks from large-scale CLIP-Seq data. Nucleic Acids Res. 42(Database issue): D92–D97. 2014. View Article : Google Scholar
12	Agarwal V, Bell GW, Nam JW and Bartel DP: Predicting effective microRNA target sites in mammalian mRNAs. Elife. 4:2015. View Article : Google Scholar
13	Tlemsani C, Pecuchet N, Gruber A, Laurendeau I, Danel C, Riquet M, Le Pimpec-Barthes F, Fabre E, Mansuet-Lupo A, Damotte D, et al: NF1 mutations identify molecular and clinical subtypes of lung adenocarcinomas. Cancer Med. 8:4330–4337. 2019.PubMed/NCBI
14	Brems H, Park C, Maertens O, Pemov A, Messiaen L, Upadhyaya M, Claes K, Beert E, Peeters K, Mautner V, et al: Glomus tumors in neurofibromatosis type 1: Genetic, functional, and clinical evidence of a novel association. Cancer Res. 69:7393–7401. 2009. View Article : Google Scholar : PubMed/NCBI
15	Stites EC, Trampont PC, Haney LB, Walk SF and Ravichandran KS: Cooperation between Noncanonical Ras Network Mutations. Cell Rep. 10:307–316. 2015. View Article : Google Scholar : PubMed/NCBI
16	Wang X, Min S, Liu H, Wu N, Liu X, Wang T, Li W, Shen Y, Wang H, Qian Z, et al: Nf1 loss promotes Kras-driven lung adenocarcinoma and results in Psat1-mediated glutamate dependence. EMBO Mol Med. 11:e98562019. View Article : Google Scholar
17	Redig AJ, Capelletti M, Dahlberg SE, Sholl LM, Mach S, Fontes C, Shi Y, Chalasani P and Jänne PA: Clinical and molecular characteristics of NF1-mutant lung cancer. Clin Cancer Res. 22:3148–3156. 2016. View Article : Google Scholar : PubMed/NCBI
18	Modak JM, Roy-O'Reilly M, Zhu L, Staff I and McCullough LD: Differential MicroRibonucleic acid expression in cardioembolic stroke. J Stroke Cerebrovasc Dis. 28:121–124. 2019. View Article : Google Scholar
19	Kanehisa M, Sato Y, Furumichi M, Morishima K and Tanabe M: New approach for understanding genome variations in KEGG. Nucleic Acids Res. 47:D590–D595. 2019. View Article : Google Scholar :
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
21	Zhang Y, Guo L, Li Y, Feng GH, Teng F, Li W and Zhou Q: MicroRNA-494 promotes cancer progression and targets adenomatous polyposis coli in colorectal cancer. Mol Cancer. 17:12018. View Article : Google Scholar : PubMed/NCBI
22	Ji Y, Han Z, Shao L and Zhao Y: Evaluation of in vivo antitumor effects of low-frequency ultrasound-mediated miRNA-133a microbubble delivery in breast cancer. Cancer Med. 5:2534–2543. 2016. View Article : Google Scholar : PubMed/NCBI
23	Wei S, Peng L, Yang J, Sang H, Jin D, Li X, Chen M, Zhang W, Dang Y and Zhang G: Exosomal transfer of miR-15b-3p enhances tumorigenesis and malignant transformation through the DYNLT1/Caspase-3/Caspase-9 signaling pathway in gastric cancer. J Exp Clin Cancer Res. 39:322020. View Article : Google Scholar : PubMed/NCBI
24	Huang XY, Huang ZL, Huang J, Xu B, Huang XY, Xu YH, Zhou J and Tang ZY: Exosomal circRNA-100338 promotes hepatocellular carcinoma metastasis via enhancing invasiveness and angiogenesis. J Exp Clin Cancer Res. 39:202020. View Article : Google Scholar : PubMed/NCBI
25	Wu Y, Liu H, Shi X, Yao Y, Yang W and Song Y: The long non-coding RNA HNF1A-AS1 regulates proliferation and metastasis in lung adenocarcinoma. Oncotarget. 6:9160–9172. 2015. View Article : Google Scholar : PubMed/NCBI
26	Thery C, Witwer KW, Aikawa E, Alcaraz MJ, Anderson JD, Andriantsitohaina R, Antoniou A, Arab T, Archer F, Atkin-Smith GK, et al: Minimal information for studies of extra-cellular vesicles 2018 (MISEV2018): A position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles. 7:15357502018. View Article : Google Scholar
27	Kalluri R: The biology and function of fibroblasts in cancer. Nat Rev Cancer. 16:582–598. 2016. View Article : Google Scholar : PubMed/NCBI
28	Ab Razak NS, Ab Mutalib NS, Mohtar MA and Abu N: Impact of chemotherapy on extracellular vesicles: Understanding the chemo-EVs. Front Oncol. 9:11132019. View Article : Google Scholar : PubMed/NCBI
29	Baffa R, Fassan M, Volinia S, O'Hara B, Liu CG, Palazzo JP, Gardiman M, Rugge M, Gomella LG, Croce CM and Rosenberg A: MicroRNA expression profiling of human metastatic cancers identifies cancer gene targets. J Pathol. 219:214–221. 2009. View Article : Google Scholar : PubMed/NCBI
30	Qian B, Wang DM, Gu XS, Zhou K, Wu J, Zhang CY and He XY: LncRNA H19 serves as a ceRNA and participates in non-small cell lung cancer development by regulating microRNA-107. Eur Rev Med Pharmacol Sci. 22:5946–5953. 2018.PubMed/NCBI
31	Philpott C, Tovell H, Frayling IM, Cooper DN and Upadhyaya M: The NF1 somatic mutational landscape in sporadic human cancers. Hum Genomics. 11:132017. View Article : Google Scholar : PubMed/NCBI
32	Rahman MA, Barger JF, Lovat F, Gao M, Otterson GA and Nana-Sinkam P: Lung cancer exosomes as drivers of epithelial mesenchymal transition. Oncotarget. 7:54852–54866. 2016. View Article : Google Scholar : PubMed/NCBI
33	You J, Li M, Cao LM, Gu QH, Deng PB, Tan Y and Hu CP: Snail1-dependent cancer-associated fibroblasts induce epithelial-mesenchymal transition in lung cancer cells via exosomes. QJM. 112:581–590. 2019. View Article : Google Scholar : PubMed/NCBI
34	He S, Li Z, Yu Y, Zeng Q, Cheng Y, Ji W, Xia W and Lu S: Exosomal miR-499a-5p promotes cell proliferation, migration and EMT via mTOR signaling pathway in lung adenocarcinoma. Exp Cell Res. 379:203–213. 2019. View Article : Google Scholar : PubMed/NCBI
35	Verset L, Tommelein J, Moles Lopez X, Decaestecker C, Boterberg T, De Vlieghere E, Salmon I, Mareel M, Bracke M, De Wever O and Demetter P: Impact of neoadjuvant therapy on cancer-associated fibroblasts in rectal cancer. Radiother Oncol. 116:449–454. 2015. View Article : Google Scholar : PubMed/NCBI
36	Wei M, Yang T, Chen X, Wu Y, Deng X, He W, Yang J and Wang Z: Malignant ascites-derived exosomes promote proliferation and induce carcinoma-associated fibroblasts transition in peritoneal mesothelial cells. Oncotarget. 8:42262–42271. 2017. View Article : Google Scholar : PubMed/NCBI
37	Zheng X, Bahr M and Doeppner TR: From tumor metastasis towards cerebral ischemia-extracellular vesicles as a general concept of intercellular communication processes. Int J Mol Sci. 20:E59952019. View Article : Google Scholar : PubMed/NCBI
38	Ning X, Zhang H, Wang C and Song X: Exosomes released by gastric cancer cells induce transition of pericytes into cancer-associated fibroblasts. Med Sci Monit. 24:2350–2359. 2018. View Article : Google Scholar : PubMed/NCBI