Exosomal microRNAs as potential biomarkers for cancer  cell migration and prognosis in hepatocellular carcinoma patient-derived cell models

Yu,Ling-Xiang; Zhang,Bo-Lun; Yang,Yuan; Wang,Meng-Chao; Lei,Guang-Lin; Gao,Yuan; Liu,Hu; Xiao,Chao-Hui; Xu,Jia-Jia; Qin,Hao; Xu,Xiao-Ya; Chen,Zi-Shuo; Zhang,Da-Dong; Li,Fu-Gen; Zhang,Shao-Geng; Liu,Rong

doi:10.3892/or.2018.6829

Exosomal microRNAs as potential biomarkers for cancer cell migration and prognosis in hepatocellular carcinoma patient-derived cell models

Authors:
- Ling-Xiang Yu
- Bo-Lun Zhang
- Yuan Yang
- Meng-Chao Wang
- Guang-Lin Lei
- Yuan Gao
- Hu Liu
- Chao-Hui Xiao
- Jia-Jia Xu
- Hao Qin
- Xiao-Ya Xu
- Zi-Shuo Chen
- Da-Dong Zhang
- Fu-Gen Li
- Shao-Geng Zhang
- Rong Liu
View Affiliations

Affiliations: Department of Surgical Oncology, Chinese People's Liberation Army (PLA) General Hospital, Beijing 100853, P.R. China, Department of General Surgery, Clinical Medical College of Weifang Medical University, Weifang, Shandong 261053, P.R. China, The Third Department of Hepatic Surgery, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, P.R. China, Department of Hepatobiliary Surgery, Hospital 302 of the PLA, Beijing 100039, P.R. China, Research and Development Institute of Precision Medicine, 3D Medicine Inc., Shanghai 201114, P.R. China
Published online on: October 29, 2018 https://doi.org/10.3892/or.2018.6829
Pages: 257-269
Copyright: © Yu 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

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide, and current treatments exhibit limited efficacy against advanced HCC. The majority of cancer-related deaths are caused by metastasis from the primary tumor, which indicates the importance of identifying clinical biomarkers for predicting metastasis and indicating prognosis. Patient-derived cells (PDCs) may be effective models for biomarker identification. In the present study, a wound healing assay was used to obtain 10 fast-migrated and 10 slow-migrated PDC cultures from 36 HCC samples. MicroRNA (miRNA) signatures in PDCs and PDC-derived exosomes were profiled by microRNA-sequencing. Differentially expressed miRNAs between the low- and fast-migrated groups were identified and further validated in 372 HCC profiles from The Cancer Genome Atlas (TCGA). Six exosomal miRNAs were identified to be differentially expressed between the two groups. In the fast-migrated group, five miRNAs (miR-140-3p, miR-30d-5p, miR-29b-3p, miR-130b-3p and miR-330-5p) were downregulated, and one miRNA (miR-296-3p) was upregulated compared with the slow-migrated group. Pathway analysis demonstrated that the target genes of the differentially expressed miRNAs were significantly enriched in the ‘focal adhesion’ pathway, which is consistent with the roles of these miRNAs in tumor metastasis. Three miRNAs, miR-30d, miR-140 and miR-29b, were significantly associated with patient survival. These findings indicated that these exosomal miRNAs may be candidate biomarkers for predicting HCC cell migration and prognosis and may guide the treatment of advanced HCC.

View Figures

View References

1	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
2	Steeg PS: Targeting metastasis. Nat Rev Cancer. 16:201–218. 2016. View Article : Google Scholar : PubMed/NCBI
3	Xu G, Zhang Y, Wei J, Jia W, Ge Z, Zhang Z and Liu X: MicroRNA-21 promotes hepatocellular carcinoma HepG2 cell proliferation through repression of mitogen-activated protein kinase-kinase 3. BMC Cancer. 13:4692013. View Article : Google Scholar : PubMed/NCBI
4	Yan LX, Wu QN, Zhang Y, Li YY, Liao DZ, Hou JH, Fu J, Zeng MS, Yun JP, Wu QL, et al: Knockdown of miR-21 in human breast cancer cell lines inhibits proliferation, in vitro migration and in vivo tumor growth. Breast Cancer Res. 13:R22011. View Article : Google Scholar : PubMed/NCBI
5	Li J, Liu K, Liu Y, Xu Y, Zhang F, Yang H, Liu J, Pan T, Chen J, Wu M, et al: Exosomes mediate the cell-to-cell transmission of IFN-α-induced antiviral activity. Nat Immunol. 14:793–803. 2013. View Article : Google Scholar : PubMed/NCBI
6	Lässer C, Alikhani VS, Ekström K, Eldh M, Paredes PT, Bossios A, Sjöstrand M, Gabrielsson S, Lötvall J and Valadi H: Human saliva, plasma and breast milk exosomes contain RNA: Uptake by macrophages. J Transl Med. 9:92011. View Article : Google Scholar : PubMed/NCBI
7	Suetsugu A, Honma K, Saji S, Moriwaki H, Ochiya T and Hoffman RM: Imaging exosome transfer from breast cancer cells to stroma at metastatic sites in orthotopic nude-mouse models. Adv Drug Deliv Rev. 65:383–390. 2013. View Article : Google Scholar : PubMed/NCBI
8	Peinado H, Alečković M, Lavotshkin S, Matei I, Costa-Silva B, Moreno-Bueno G, Hergueta-Redondo M, Williams C, García-Santos G, Ghajar C, et al: Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET. Nat Med. 18:883–891. 2012. View Article : Google Scholar : PubMed/NCBI
9	Kogure T and Patel T: Isolation of extracellular nanovesicle microRNA from liver cancer cells in culture. Methods Mol Biol. 1024:11–18. 2013. View Article : Google Scholar : PubMed/NCBI
10	van Staveren WC, Solís DY, Hébrant A, Detours V, Dumont JE and Maenhaut C: Human cancer cell lines: Experimental models for cancer cells in situ? For cancer stem cells? Biochim Biophys Acta. 1795:92–103. 2009.PubMed/NCBI
11	Byrne AT, Alférez DG, Amant F, Annibali D, Arribas J, Biankin AV, Bruna A, Budinská E, Caldas C, Chang DK, et al: Interrogating open issues in cancer precision medicine with patient-derived xenografts. Nat Rev Cancer. 17:254–268. 2017. View Article : Google Scholar : PubMed/NCBI
12	Buitrago DH, Patnaik SK, Kadota K, Kannisto E, Jones DR and Adusumilli PS: Small RNA sequencing for profiling microRNAs in long-term preserved formalin-fixed and paraffin-embedded non-small cell lung cancer tumor specimens. PLoS One. 10:e01215212015. View Article : Google Scholar : PubMed/NCBI
13	Huang J, Qin H, Yang Y, Chen X, Zhang J, Laird S, Wang CC, Chan TF and Li TC: A comparison of transcriptomic profiles in endometrium during window of implantation between women with unexplained recurrent implantation failure and recurrent miscarriage. Reproduction. 153:749–758. 2017. View Article : Google Scholar : PubMed/NCBI
14	van Roosmalen W, Le Dévédec SE, Golani O, Smid M, Pulyakhina I, Timmermans AM, Look MP, Zi D, Pont C, de Graauw M, et al: Tumor cell migration screen identifies SRPK1 as breast cancer metastasis determinant. J Clin Invest. 125:1648–1664. 2015. View Article : Google Scholar : PubMed/NCBI
15	Rodríguez M, Bajo-Santos C, Hessvik NP, Lorenz S, Fromm B, Berge V, Sandvig K, Linē A and Llorente A: Identification of non-invasive miRNAs biomarkers for prostate cancer by deep sequencing analysis of urinary exosomes. Mol Cancer. 16:1562017. View Article : Google Scholar : PubMed/NCBI
16	Baglio SR, Rooijers K, Koppers-Lalic D, Verweij FJ, Pérez Lanzón M, Zini N, Naaijkens B, Perut F, Niessen HW, Baldini N, et al: Human bone marrow- and adipose-mesenchymal stem cells secrete exosomes enriched in distinctive miRNA and tRNA species. Stem Cell Res Ther. 6:1272015. View Article : Google Scholar : PubMed/NCBI
17	Tang YT, Huang YY, Zheng L, Qin SH, Xu XP, An TX, Xu Y, Wu YS, Hu XM, Ping BH, et al: Comparison of isolation methods of exosomes and exosomal RNA from cell culture medium and serum. Int J Mol Med. 40:834–844. 2017. View Article : Google Scholar : PubMed/NCBI
18	Gernapudi R, Yao Y, Zhang Y, Wolfson B, Roy S, Duru N, Eades G, Yang P and Zhou Q: Targeting exosomes from preadipocytes inhibits preadipocyte to cancer stem cell signaling in early-stage breast cancer. Breast Cancer Res Treat. 150:685–695. 2015. View Article : Google Scholar : PubMed/NCBI
19	Chen D, Guo W, Qiu Z, Wang Q, Li Y, Liang L, Liu L, Huang S, Zhao Y and He X: MicroRNA-30d-5p inhibits tumour cell proliferation and motility by directly targeting CCNE2 in non-small cell lung cancer. Cancer Lett. 362:208–217. 2015. View Article : Google Scholar : PubMed/NCBI
20	Nishikawa R, Chiyomaru T, Enokida H, Inoguchi S, Ishihara T, Matsushita R, Goto Y, Fukumoto I, Nakagawa M and Seki N: Tumour-suppressive microRNA-29s directly regulate LOXL2 expression and inhibit cancer cell migration and invasion in renal cell carcinoma. FEBS Lett. 589:2136–2145. 2015. View Article : Google Scholar : PubMed/NCBI
21	Shui Y, Yu X, Duan R, Bao Q, Wu J, Yuan H and Ma C: miR-130b-3p inhibits cell invasion and migration by targeting the Notch ligand Delta-like 1 in breast carcinoma. Gene. 609:80–87. 2017. View Article : Google Scholar : PubMed/NCBI
22	Su BB, Zhou SW, Gan CB and Zhang XN: MiR-330-5p regulates tyrosinase and PDIA3 expression and suppresses cell proliferation and invasion in cutaneous malignant melanoma. J Surg Res. 203:434–440. 2016. View Article : Google Scholar : PubMed/NCBI
23	Zhou L, Liu F, Wang X and Ouyang G: The roles of microRNAs in the regulation of tumor metastasis. Cell Biosci. 5:322015. View Article : Google Scholar : PubMed/NCBI
24	Gaziel-Sovran A, Segura MF, Di Micco R, Collins MK, Hanniford D, Vega-Saenz de Miera E, Rakus JF, Dankert JF, Shang S, Kerbel RS, et al: miR-30b/30d regulation of GalNAc transferases enhances invasion and immunosuppression during metastasis. Cancer Cell. 20:104–118. 2011. View Article : Google Scholar : PubMed/NCBI
25	He Z, Yu L, Luo S, Li M, Li J, Li Q, Sun Y and Wang C: miR-296 inhibits the metastasis and epithelial-mesenchymal transition of colorectal cancer by targeting S100A4. BMC Cancer. 17:1402017. View Article : Google Scholar : PubMed/NCBI
26	Panera N, Crudele A, Romito I, Gnani D and Alisi A: Focal adhesion kinase: Insight into molecular roles and functions in hepatocellular carcinoma. Int J Mol Sci. 18:992017.doi: 10.3390/ijms18010099. View Article : Google Scholar :