MicroRNA‑24‑1‑5p promotes malignant melanoma cell autophagy and apoptosis via regulating ubiquitin D

Xiao,Yujuan; Diao,Qingchun; Liang,Yanhua; Peng,Yusheng; Zeng,Kang

doi:10.3892/mmr.2017.7614

MicroRNA‑24‑1‑5p promotes malignant melanoma cell autophagy and apoptosis via regulating ubiquitin D

Authors:
- Yujuan Xiao
- Qingchun Diao
- Yanhua Liang
- Yusheng Peng
- Kang Zeng
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Affiliations: Department of Dermatology, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Department of Dermatology, Chongqing Traditional Chinese Medicine Hospital, Chongqing 400011, P.R. China, Department of Dermatology, Shenzhen Hospital, Southern Medical University, Shenzhen, Guangdong 518100, P.R. China
Published online on: September 26, 2017 https://doi.org/10.3892/mmr.2017.7614
Pages: 8448-8454

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Abstract

The present study aimed to investigate the key roles and possible regulatory mechanism of microRNA (miR)‑24‑1‑5p in regulating the autophagy, and apoptosis of malignant melanoma cells. The expression levels of miR‑24‑1‑5p in malignant melanoma tissues were determined. Human melanoma A375 cells were transfected with miR‑24‑1‑5p mimic and control. The effects of miR‑24‑1‑5p overexpression on regulating the expressions of autophagy‑related proteins [microtubule‑associated protein 1A/1B‑light chain 3 (LC3)‑II, LC3‑I and Beclin‑1] and apoptosis‑related proteins [apoptosis regulator Bcl‑2 (Bcl‑2) and (BCL2 like 1) Bcl‑xL] were investigated. The percentage of apoptotic cells in different transfected cells was detected. In addition, luciferase reporter assays were performed to confirm whether ubiquitin D (UBD) was a target of miR‑24‑1‑5p. The effects of UBD silencing on autophagy and apoptosis were also investigated. The expression levels of janus kinase (JNK), phosphorylated (P)‑JNK, Jun proto‑oncogene AP‑1 transcription factor subunit (c‑Jun) and p‑c‑Jun were determined following the overexpression of miR‑24‑1‑5p, and UBD. In comparison with adjacent normal tissues, miR‑24‑1‑5p was significantly downregulated in malignant melanoma tissues. Overexpression of miR‑24‑1‑5p significantly increased the levels of LC3‑II/I ratio and Beclin‑1 expression, and decreased the expression levels of Bcl‑2 and Bcl‑xL. Flow cytometry also showed that miR‑24‑1‑5p overexpression promoted cell apoptosis. Moreover, UBD was confirmed as a direct target of miR‑24‑1‑5p. Silencing of UBD promoted melanoma cell autophagy and apoptosis via regulating the expression levels of related proteins. Besides, the levels of the p‑JNK/JNK and p‑c‑Jun/Jun ratios were significantly increased following miR‑24‑1‑5p overexpression, which were reversed following co‑overexpression of miR‑24‑1‑5p, and UBD. Overexpression of miR‑24‑1‑5p may target UBD, and subsequently promote the autophagy and apoptosis of malignant melanoma cells through activation of the JNK signaling pathway.

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1	Gray-Schopfer V, Wellbrock C and Marais R: Melanoma biology and new targeted therapy. Nature. 445:851–857. 2007. View Article : Google Scholar : PubMed/NCBI
2	Luna JM, Hernández Guerrero A, Méndez R Romero and Ortiz JL Luviano: Solution of the inverse bio-heat transfer problem for a simplified dermatological application: Case of skin cancer. Ingenierí Mecáni Tecnolog Desarroll. 4:2014.
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	Bloethner S, Scherer D, Drechsel M, Hemminki K and Kumar R: Malignant melanoma-a genetic overview. Actas Dermosifiliogr. 100 Suppl 1:S38–S51. 2009. View Article : Google Scholar
5	Gogas HJ, Kirkwood JM and Sondak VK: Chemotherapy for metastatic melanoma: Time for a change? Cancer. 109:455–464. 2007. View Article : Google Scholar : PubMed/NCBI
6	Liu L, Kritsanida M, Magiatis P, Gaboriaud N, Wang Y, Wu J, Buettner R, Yang F, Nam S, Skaltsounis L and Jove R: A novel 7-bromoindirubin with potent anticancer activity suppresses survival of human melanoma cells associated with inhibition of STAT3 and Akt signaling. Cancer Biol Ther. 13:1255–1261. 2012. View Article : Google Scholar : PubMed/NCBI
7	Glud M and Gniadecki R: MicroRNAs in the pathogenesis of malignant melanoma. J Eur Acad Dermatol Venereol. 27:142–150. 2013. View Article : Google Scholar : PubMed/NCBI
8	Völler D, Ott C and Bosserhoff A: MicroRNAs in malignant melanoma. Clin Biochem. 46:909–917. 2013. View Article : Google Scholar : PubMed/NCBI
9	Hanahan D and Weinberg RA: Hallmarks of cancer: The next generation. Cell. 144:646–674. 2011. View Article : Google Scholar : PubMed/NCBI
10	Maiuri MC, Zalckvar E, Kimchi A and Kroemer G: Self-eating and self-killing: Crosstalk between autophagy and apoptosis. Nat Rev Mol Cell Biol. 8:741–752. 2007. View Article : Google Scholar : PubMed/NCBI
11	Xu J, Wang Y, Tan X and Jing H: MicroRNAs in autophagy and their emerging roles in crosstalk with apoptosis. Autophagy. 8:873–882. 2012. View Article : Google Scholar : PubMed/NCBI
12	Satzger I, Mattern A, Kuettler U, Weinspach D, Niebuhr M, Kapp A and Gutzmer R: microRNA-21 is upregulated in malignant melanoma and influences apoptosis of melanocytic cells. Exp Dermatol. 21:509–514. 2012. View Article : Google Scholar : PubMed/NCBI
13	Bhattacharya A, Schmitz U, Raatz Y, Schönherr M, Kottek T, Schauer M, Franz S, Saalbach A, Anderegg U, Wolkenhauer O, et al: miR-638 promotes melanoma metastasis and protects melanoma cells from apoptosis and autophagy. Oncotarget. 6:2966–2980. 2015. View Article : Google Scholar : PubMed/NCBI
14	Singleton PA, Mirzapoiazova T, Mambetsariev N, Ge L, Mambetsariev B, Lennon FE, Poroyko V, Fang Y, Mutlu GM and Siddiqui S: miR-24-1-5p regulates high molecular weight Hyaluronan (hmw-Ha)-mediated enhancement of vascular Integrity: Role of foxa3. Am J Respir Crit Care Med. 191:A26462015.
15	Singleton PA, Siddiqui SS, Mirzapoiazova T, Mambetsariev N, Mambetsariev B and Lennon FE: MicroRNA Hsa-miR-24-1* regulates high molecular weight hyaluronan-mediated human pulmonary endothelial barrier enhancement. Am J Respir Crit Care Med. 183:A41732011.
16	Sand M, Skrygan M, Sand D, Georgas D, Gambichler T, Hahn SA, Altmeyer P and Bechara FG: Comparative microarray analysis of microRNA expression profiles in primary cutaneous malignant melanoma, cutaneous malignant melanoma metastases, and benign melanocytic nevi. Cell Tissue Res. 351:85–98. 2013. View Article : Google Scholar : PubMed/NCBI
17	Hooi WC: The role of ROS-mediated ERK and JNK activation in the induction of autophagy and apoptosis in tumour cells by a novel small molecule compound. Ph D2009
18	Hassan M, Watari H, Abualmaaty A, Ohba Y and Sakuragi N: Apoptosis and molecular targeting therapy in cancer. Biomed Res Int. 2014:1508452014. View Article : Google Scholar : PubMed/NCBI
19	Sankari SL, Masthan KM, Babu NA, Bhattacharjee T and Elumalai M: Apoptosis in cancer-an update. Asian Pac J Cancer Prev. 13:4873–4878. 2012. View Article : Google Scholar : PubMed/NCBI
20	Leiter U, Schmid RM, Kaskel P, Peter RU and Krähn G: Antiapoptotic bcl-2 and bcl-xL in advanced malignant melanoma. Arch Dermatol Res. 292:225–232. 2000. View Article : Google Scholar : PubMed/NCBI
21	Wacheck V, Losert D, Günsberg P, Vornlocher HP, Hadwiger P, Geick A, Pehamberger H, Müller M and Jansen B: Small interfering RNA targeting bcl-2 sensitizes malignant melanoma. Oligonucleotides. 13:393–400. 2003. View Article : Google Scholar : PubMed/NCBI
22	Maycotte P and Thorburn A: Autophagy and cancer therapy. Cancer Biol Ther. 11:127–137. 2011. View Article : Google Scholar : PubMed/NCBI
23	Janku F, Mcconkey DJ, Hong DS and Kurzrock R: Autophagy as a target for anticancer therapy. Nat Rev Clin Oncol. 8:528–539. 2011. View Article : Google Scholar : PubMed/NCBI
24	Sivridis E, Koukourakis MI, Mendrinos SE, Karpouzis A, Fiska A, Kouskoukis C and Giatromanolaki A: Beclin-1 and LC3A expression in cutaneous malignant melanomas: A biphasic survival pattern for beclin-1. Melanoma Res. 21:188–195. 2011. View Article : Google Scholar : PubMed/NCBI
25	Yuan-Ting SU, Jin HL and Dermatology DO: The advances in Beclin 1 and MAP1LC3 in malignant melanoma. Chin J Dermatovenereol. 2014.
26	Jian X, Xiao-yan Z, Bin H, Yu-feng Z, Bo K, Zhi-nong W and Xin N: miR-204 regulate cardiomyocyte autophagy induced by hypoxia-reoxygenation through LC3-II. Int J Cardiol. 148:110–112. 2011. View Article : Google Scholar : PubMed/NCBI
27	Oliva J, Bardaggorce F, French BA, Li J, McPhaul L, Amidi F, Dedes J, Habibi A, Nguyen S and French SW: Fat10 is an epigenetic marker for liver preneoplasia in a drug-primed mouse model of tumorigenesis. Exp Mol Pathol. 84:102–112. 2008. View Article : Google Scholar : PubMed/NCBI
28	Ji F, Jin X, Jiao CH, Xu QW, Wang ZW and Chen YL: FAT10 level in human gastric cancer and its relation with mutant p53 level, lymph node metastasis and TNM staging. World J Gastroenterol. 15:2228–2233. 2009. View Article : Google Scholar : PubMed/NCBI
29	Yan DW, Li DW, Yang YX, Xia J, Wang XL, Zhou CZ, Fan JW, Wen YG, Sun HC, Wang Q, et al: Ubiquitin D is correlated with colon cancer progression and predicts recurrence for stage II–III disease after curative surgery. Br J Cancer. 103:961–969. 2010. View Article : Google Scholar : PubMed/NCBI
30	Brozzi F, Gerlo S, Grieco FA, Juusola M, Balhuizen A, Lievens S, Gysemans C, Bugliani M, Mathieu C, Marchetti P, et al: Ubiquitin D regulates IRE1α/c-Jun N-terminal kinase (JNK) protein-dependent apoptosis in pancreatic beta cells. J Biol Chem. 291:12040–12056. 2016. View Article : Google Scholar : PubMed/NCBI
31	Wang Y, Xu L, Lou LL, Song SJ, Yao GD, Ge MY, Hayashi T, Tashiro SI, Onodera S and Ikejima T: Timosaponin AIII induces apoptosis and autophagy in human melanoma A375-S2 cells. Arch Pharm Res. 40:69–78. 2017. View Article : Google Scholar : PubMed/NCBI