Overexpression of microRNA‑21 inhibits the growth and metastasis of melanoma cells by targeting MKK3

Zhou,Meng; Yu,Xiaoqian; Jing,Zhenhai; Wu,Wei; Lu,Chenglong

doi:10.3892/mmr.2019.10408

Overexpression of microRNA‑21 inhibits the growth and metastasis of melanoma cells by targeting MKK3

Authors:
- Meng Zhou
- Xiaoqian Yu
- Zhenhai Jing
- Wei Wu
- Chenglong Lu
View Affiliations

Affiliations: Department of Dermatology, Qilu Hospital of Shandong University, Qingdao, Shandong 266000, P.R. China, Department of Dermatology, Qingdao Hiser Medical Group, Qingdao Hospital of Traditional Chinese Medicine, Qingdao, Shandong 266032, P.R. China, Department of Oncology, Qingdao Hiser Medical Group, Qingdao Hospital of Traditional Chinese Medicine, Qingdao, Shandong 266032, P.R. China, College of Food Science and Technology, Qingdao Agricultural University, Qingdao, Shandong 266179, P.R. China, Department of Emergency, Qilu Hospital of Shandong University, Qingdao, Shandong 266000, P.R. China
Published online on: June 20, 2019 https://doi.org/10.3892/mmr.2019.10408
Pages: 1797-1807
Copyright: © Zhou 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

Melanoma is an aggressive skin carcinoma with poor prognosis, and is prevalent worldwide. It was demonstrated that microRNA (miR)‑21 and mitogen‑activated protein kinase kinase 3 (MKK3) both participated in the occurrence and development of various tumors; however, their detailed roles in the progression of melanoma remain unclear. Reverse transcription‑quantitative PCR (RT‑qPCR) and western blot analyses were conducted to examine the expression levels of miR‑21 and MKK3 in clinical specimens of patients with melanoma and melanoma cell lines. A dual‑luciferase reporter assay was performed to verify the target interaction between miR‑21 and MKK3. The mRNA and protein expressions of MKK3 were measured using RT‑qPCR and western blot analysis, respectively, following transfection with miR‑21 mimics and inhibitor. Subsequently, Cell Counting Kit‑8 and colony formation assays, and flow cytometry were conducted to assess the effects of miR‑21 and MKK3 on the cell growth of melanoma. Cell migration and invasion experiments were performed to evaluate the effects of miR‑21 and MKK3 on the cell metastasis of melanoma. It was revealed that MKK3 was upregulated, and miR‑21 was downregulated in patients with melanoma and melanoma cell lines. MKK3 was demonstrated to be a direct target of miR‑21. Furthermore, it was demonstrated that upregulated miR‑21 expression and downregulated MKK3 expression suppressed cell proliferation and colony formation, promoted apoptosis, delayed the cell cycle, and inhibited cell migration and invasion. The present findings suggested that miR‑21 could inhibit the cell growth and metastasis of melanoma by negatively regulating MKK3.

View Figures

View References

1	Owens B: Melanoma. Nature. 515 (Suppl):S1092014. View Article : Google Scholar : PubMed/NCBI
2	Mohammadpour A, Derakhshan M, Darabi H, Hedayat P and Momeni M: Melanoma: Where we are and where we go. J Cell Physiol. 234:3307–3320. 2019. View Article : Google Scholar : PubMed/NCBI
3	Little EG and Eide MJ: Update on the current state of melanoma incidence. Dermatol Clin. 30:355–361. 2012. View Article : Google Scholar : PubMed/NCBI
4	Ko JS: The immunology of melanoma. Clin Lab Med. 37:449–471. 2017. View Article : Google Scholar : PubMed/NCBI
5	Hayward NK, Wilmott JS, Waddell N, Johansson PA, Field MA, Nones K, Patch AM, Kakavand H, Alexandrov LB, Burke H, et al: Whole-genome landscapes of major melanoma subtypes. Nature. 545:175–180. 2017. View Article : Google Scholar : PubMed/NCBI
6	Wong CW, Fan YS, Chan TL, Chan AS, Ho LC, Ma TK, Yuen ST and Leung SY; Cancer Genome Project, : BRAF and NRAS mutations are uncommon in melanomas arising in diverse internal organs. J Clin Pathol. 58:640–644. 2005. View Article : Google Scholar : PubMed/NCBI
7	Mohr AM and Mott JL: Overview of microRNA biology. Semin Liver Dis. 35:3–11. 2015. View Article : Google Scholar : PubMed/NCBI
8	Ha M and Kim VN: Regulation of microRNA biogenesis. Nat Rev Mol Cell Biol. 15:509–524. 2014. View Article : Google Scholar : PubMed/NCBI
9	Fuchs Wightman F, Giono LE, Fededa JP and de la Mata M: Target RNAs strike back on MicroRNAs. Front Genet. 9:4352018. View Article : Google Scholar : PubMed/NCBI
10	Treiber T, Treiber N and Meister G: Regulation of microRNA biogenesis and its crosstalk with other cellular pathways. Nat Rev Mol Cell Biol. 20:5–20. 2019. View Article : Google Scholar : PubMed/NCBI
11	Mumford SL, Towler BP, Pashler AL, Gilleard O, Martin Y and Newbury SF: Circulating MicroRNA biomarkers in melanoma: Tools and challenges in personalised medicine. Biomolecules. 8:E212018. View Article : Google Scholar : PubMed/NCBI
12	Mirzaei H, Gholamin S, Shahidsales S, Sahebkar A, Jaafari MR, Mirzaei HR, Hassanian SM and Avan A: MicroRNAs as potential diagnostic and prognostic biomarkers in melanoma. Eur J Cancer. 53:25–32. 2016. View Article : Google Scholar : PubMed/NCBI
13	Zhu Y, Zhang HL, Wang QY, Chen MJ and Liu LB: Overexpression of microRNA-612 restrains the growth, invasion, and tumorigenesis of melanoma cells by targeting espin. Mol Cells. 41:119–126. 2018.PubMed/NCBI
14	Zhang J, Liu WL, Zhang L, Ge R, He F, Gao TY, Tian Q, Mu X, Chen LH, Chen W and Li X: MiR-637 suppresses melanoma progression through directly targeting P-REX2a and inhibiting PTEN/AKT signaling pathway. Cell Mol Biol (Noisy-le-grand). 64:50–57. 2018. View Article : Google Scholar : PubMed/NCBI
15	Panza E, Ercolano G, De Cicco P, Armogida C, Scognamiglio G, Botti G, Cirino G and Ianaro A: MicroRNA-143-3p inhibits growth and invasiveness of melanoma cells by targeting cyclooxygenase-2 and inversely correlates with malignant melanoma progression. Biochem Pharmacol. 156:52–59. 2018. View Article : Google Scholar : PubMed/NCBI
16	Pfeffer SR, Yang CH and Pfeffer LM: The role of miR-21 in cancer. Drug Dev Res. 76:270–277. 2015. View Article : Google Scholar : PubMed/NCBI
17	Zhang H, Li J, Li G and Wang S: Effects of celastrol on enhancing apoptosis of ovarian cancer cells via the downregulation of microRNA21 and the suppression of the PI3K/Akt-NF-κB signaling pathway in an in vitro model of ovarian carcinoma. Mol Med Rep. 14:5363–5368. 2016. View Article : Google Scholar : PubMed/NCBI
18	Mu Z and Sun Q: Cantharidin inhibits melanoma cell proliferation via the miR21mediated PTEN pathway. Mol Med Rep. 18:4603–4610. 2018.PubMed/NCBI
19	Wandler A, Riber-Hansen R, Hager H, Hamilton-Dutoit SJ, Schmidt H, Nielsen BS, Stougaard M and Steiniche T: Quantification of microRNA-21 and microRNA-125b in melanoma tissue. Melanoma Res. 27:417–428. 2017. View Article : Google Scholar : PubMed/NCBI
20	Mao XH, Chen M, Wang Y, Cui PG, Liu SB and Xu ZY: MicroRNA-21 regulates the ERK/NF-κB signaling pathway to affect the proliferation, migration, and apoptosis of human melanoma A375 cells by targeting SPRY1, PDCD4, and PTEN. Mol Carcinog. 56:886–894. 2017. View Article : Google Scholar : PubMed/NCBI
21	Zhang HL, Si LB, Zeng A, Long F, Qi Z, Zhao R and Bai M: MicroRNA-21 antisense oligonucleotide improves the sensitivity of human melanoma cells to cisplatin: An in vitro study. J Cell Biochem. 119:3129–3141. 2018. View Article : Google Scholar : PubMed/NCBI
22	Bossi G: MKK3 as oncotarget. Aging (Albany NY). 8:1–2. 2016. View Article : Google Scholar : PubMed/NCBI
23	Huth HW, Albarnaz JD, Torres AA, Bonjardim CA and Ropert C: MEK2 controls the activation of MKK3/MKK6-p38 axis involved in the MDA-MB-231 breast cancer cell survival: Correlation with cyclin D1 expression. Cell Signal. 28:1283–1291. 2016. View Article : Google Scholar : PubMed/NCBI
24	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 : PubMed/NCBI
25	Grimson A, Farh KK, Johnston WK, Garrett-Engele P, Lim LP and Bartel DP: MicroRNA targeting specificity in mammals: Determinants beyond seed pairing. Mol Cell. 27:91–105. 2007. View Article : Google Scholar : PubMed/NCBI
26	Fattouh K, Ducroux E, Decullier E, Kanitakis J, Morelon E, Boissonnat P, Sebbag L, Jullien D and Euvrard S: Increasing incidence of melanoma after solid organ transplantation: A retrospective epidemiological study. Transpl Int. 30:1172–1180. 2017. View Article : Google Scholar : PubMed/NCBI
27	Lee JA: Current evidence about the causes of malignant melanoma. Prog Clin Cancer. 6:151–161. 1975.PubMed/NCBI
28	Wikstrom JD, Lundeberg L, Frohm-Nilsson M and Girnita A: Differences in cutaneous melanoma treatment and patient satisfaction. PLoS One. 13:e02055172018. View Article : Google Scholar : PubMed/NCBI
29	Marcell Szasz A, Malm J, Rezeli M, Sugihara Y, Betancourt LH, Rivas D, Gyorffy B and Marko-Varga G: Challenging the heterogeneity of disease presentation in malignant melanoma-impact on patient treatment. Cell Biol Toxicol. 39:1–14. 2019. View Article : Google Scholar
30	Rockberg J, Amelio JM, Taylor A, Jörgensen L, Ragnhammar P and Hansson J: Epidemiology of cutaneous melanoma in Sweden-Stage-specific survival and rate of recurrence. Int J Cancer. 139:2722–2729. 2016. View Article : Google Scholar : PubMed/NCBI
31	Baldari S, Ubertini V, Garufi A, D'Orazi G and Bossi G: Targeting MKK3 as a novel anticancer strategy: Molecular mechanisms and therapeutical implications. Cell Death Dis. 6:e16212015. View Article : Google Scholar : PubMed/NCBI
32	Wu Y, Song Y, Xiong Y, Wang X, Xu K, Han B, Bai Y, Li L, Zhang Y and Zhou L: MicroRNA-21 (Mir-21) promotes cell growth and invasion by repressing tumor suppressor PTEN in colorectal cancer. Cell Physiol Biochem. 43:945–958. 2017. View Article : Google Scholar : PubMed/NCBI
33	Lynch MP, Nawaz S and Gerschenson LE: Evidence for soluble factors regulating cell death and cell proliferation in primary cultures of rabbit endometrial cells grown on collagen. Proc Natl Acad Sci USA. 83:4784–4788. 1986. View Article : Google Scholar : PubMed/NCBI
34	Broussard L, Howland A, Ryu S, Song K, Norris D, Armstrong CA and Song PI: Melanoma cell death mechanisms. Chonnam Med J. 54:135–142. 2018. View Article : Google Scholar : PubMed/NCBI
35	Miller FR: Immune mechanisms in the sequential steps of metastasis. Crit Rev Oncog. 4:293–311. 1993.PubMed/NCBI
36	Scully OJ, Bay BH, Yip G and Yu Y: Breast cancer metastasis. Cancer Genomics Proteomics. 9:311–320. 2012.PubMed/NCBI
37	Wu J, Zhang Y, Cheng R, Gong W, Ding T, Zhai Q, Wang Y, Meng B and Sun B: Expression of epithelial-mesenchymal transition regulatorsTWIST, SLUG and SNAIL in follicular thyroid tumours may relate to widely invasive, poorly differentiated and distant metastasis. Histopathology. 74:780–791. 2019. View Article : Google Scholar : PubMed/NCBI