miR‑181a‑5p regulates the proliferation and apoptosis of glomerular mesangial cells by targeting KLF6

Liang,Xinyue; Xu,Wen

doi:10.3892/etm.2020.8780

miR‑181a‑5p regulates the proliferation and apoptosis of glomerular mesangial cells by targeting KLF6

Authors:
- Xinyue Liang
- Wen Xu
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Affiliations: Department of Geriatrics, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, P.R. China, State Key Laboratory of Bioreactor Engineering and Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai 200127, P.R. China
Published online on: May 21, 2020 https://doi.org/10.3892/etm.2020.8780
Pages: 1121-1128

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Abstract

Diabetic nephropathy (DN) is a chronic loss of kidney function that frequently occurs in patients with diabetes mellitus and is characterized by abnormal glomerular mesangial cell (GMC) proliferation and apoptosis. By using microarray analysis, microRNA (miR)‑181a‑5p has previously been identified to be dysregulated in DN. The present study aimed to determine the underlying molecular mechanisms and function of miR‑181a‑5p in GMCs under DN conditions. First, reverse transcription‑quantitative PCR was performed to detect miR‑181a‑5p and kruppel‑like factor 6 (KLF6) expression in GMCs following high‑glucose treatment. Subsequently, MTT and flow cytometric assays were performed in order to determine the effect of miR‑181a‑5p and KLF6 on high‑glucose‑driven GMC proliferation and apoptosis. After confirming that KLF6 was a target gene of miR‑181a‑5p via a bioinformatics analysis and luciferase reporter assay, the mRNA and protein expression levels of associated factors in different treatment groups were measured. The results demonstrated that miR‑181a‑5p was significantly downregulated, while KLF6 was significantly upregulated in GMCs following treatment with high glucose. Furthermore, overexpression of miR‑181a led to suppression of cell proliferation and promoted apoptosis of GMCs induced by high glucose, while these effects were inhibited by co‑transfection with KLF6. Finally, miR‑181‑5p was demonstrated to inhibit the expression of KLF6, Bcl‑2, Wnt1 and β‑catenin, while increasing the expression levels of Bax and caspase‑3. In conclusion, the expression levels of miR‑181a‑5p were downregulated in GMCs following treatment with high glucose and overexpression of miR‑181a‑5p may inhibit GMC proliferation and promote apoptosis, at least partially through targeting KLF6 via the Wnt/β‑catenin signaling pathway. Overall, the results of the present study suggest that miR‑181a‑5p may have a crucial role in the occurrence and development of DN and may be a valuable diagnostic marker and therapeutic target for DN.

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1	He L and Hannon GJ: MicroRNAs: Small RNAs with a big role in gene regulation. Nav Rev Genet. 5:522–531. 2004.PubMed/NCBI View Article : Google Scholar
2	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism and function. Cell. 116:281–297. 2004.PubMed/NCBI View Article : Google Scholar
3	Liu X, Li M, Peng Y, Hu X, Xu J, Zhu S, Yu Z and Han S: miR-30c regulates proliferation, apoptosis and differentiation via the Shh signaling pathway in P19 cells. Exp Mol Med. 48(e248)2016.PubMed/NCBI View Article : Google Scholar
4	Vidal AF, Cruz AM, Magalhães L, Pereira AL, Anaissi AK, Alves NC, Albuquerque PJ, Burbano RM, Demachki S and Ribeiro-dos-Santos Â: hsa-miR-29c and hsa-miR-135b differential expression as potential biomarker of gastric carcinogenesis. World J Gastroenterol. 22:2060–2070. 2016.PubMed/NCBI View Article : Google Scholar
5	Bansal N, Dhaliwal R and Weinstock RS: Management of diabetics in the elderly. Med Clin North Am. 99:351–377. 2015.PubMed/NCBI View Article : Google Scholar
6	Nam S, Chesla C, Stotts NA, Kroon L and Janson SL: Barriers to diabetes management: Patient and provider factors. Diabetes Res Clin Pract. 93:1–9. 2011.PubMed/NCBI View Article : Google Scholar
7	Abboud HE: Mesangial cell biology. Exp Cell Res. 318:979–985. 2012.PubMed/NCBI View Article : Google Scholar
8	Danesh FR, Sadeghi MM, Amro N, Philips C, Zeng L, Lin S, Sahai A and Kanwar YS: 3-Hydroxy-3-methylglutaryl CoA reductase inhibitors prevent high glucose-induced proliferation of mesangial cells via modulation of Rho GTPase/p21 signaling pathway: Implications for diabetic nephropathy. Proc Natl Acad Sci USA. 99:8301–8305. 2002.PubMed/NCBI View Article : Google Scholar
9	Liu ZM, Zheng HY, Chen LH, Li YL, Wang Q, Liao CF and Li XW: Low expression of miR-203 promoted diabetic nephropathy via increasing TLR4. Eur Rev Med Pharmacol Sci. 22:5627–5634. 2018.PubMed/NCBI View Article : Google Scholar
10	Zhao D, Jia J and Shao H: iR-30e targets GLIPR-2 to modulate diabetic nephropathy: In vitro and in vivo experiments. J Mol Endocrinol. 59:181–190. 2017.PubMed/NCBI View Article : Google Scholar
11	Wang J, Duan L, Gao Y, Zhou S, Liu Y, Wei S, An S, Liu J, Tian L and Wang S: Angiotensin II receptor blocker valsartan ameliorates cardiac fibrosis partly by inhibiting miR-21 expression in diabetic nephropathy mice. Mol Med Endorcrinol. 472:149–158. 2018.PubMed/NCBI View Article : Google Scholar
12	He YQ, Ding YL, Liang BY, Lin JJ, Kim TY, Yu HB, Hang HW and Wang K: A systematic study of dysregulated microRNA in type 2 diabetes mellitus. Int J Mol Sci. 18: pii(456)2017.PubMed/NCBI View Article : Google Scholar
13	Assmann TS, Recamonde-Mendoza M, De Souza BM and Crispim D: MicroRNA expression profiles and type 1 diabetes mellitus: Systematic review and bioinformatic analysis. Endocr Connect. 6:773–790. 2017.PubMed/NCBI View Article : Google Scholar
14	Tetreault MP, Yang Y and Katz JP: Krüppel-like factors in cancer. Nat Rev Cancer. 13:701–713. 2013.PubMed/NCBI View Article : Google Scholar
15	Huang X, Li X and Guo B: KLF6 induces apoptosis in prostate cancer cells through up-regulation of ATF3. J Biol Chem. 283:29795–29801. 2008.PubMed/NCBI View Article : Google Scholar
16	Wahab AH, Kassem AM, Matter S, EI Deen AF, Helmy AS, Ismaeil MM and Zakaria MS: Role of KLF6 tumor suppressor gene mutations in the development of colorectal carcinoma in an Egyptian population. Hepatogastroenterology. 57:1405–1410. 2010.PubMed/NCBI
17	Rane MJ, Zhao Y and Cai L: Krüppel-like factors (KLFs) in renal physiology and disease. EBioMedicine. 40:743–750. 2019.PubMed/NCBI View Article : Google Scholar
18	Xu S, Lv YY, Zhao J, Wang JP, Zhao X and Wang S: Inhibitory effects of Shenkang injection and its main component emodin on the proliferation of high glucose-induced renal mesangial cells through cell cycle regulation and induction of apoptosis. Mol Med Rep. 14:3381–3388. 2016.PubMed/NCBI View Article : Google Scholar
19	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.PubMed/NCBI View Article : Google Scholar
20	Li Y, Kuscu C, Banach A, Zhang Q, Pulkoski-Gross A, Kim D, Liu J, Roth E, Li E, Shroyer KR, et al: MiR-181a-5p inhibits cancer cell migration and angiogenesis via downregulation of matrix metalloproteinase-14. Cancer Res. 75:2674–2685. 2015.PubMed/NCBI View Article : Google Scholar
21	Taylor MA, Sossey-Alaoui K, Thompson CL, Danielpour D and Schiemann WP: TGF-β upregulates miR-181a expression to promote breast cancer metastasis. J Clin Invest. 123:150–163. 2013.PubMed/NCBI View Article : Google Scholar
22	Wei Y, Tao X, Xu H, Chen Y, Zhu L, Tang G, Li M, Jiang A, Shuai S, Ma J, et al: Role of miR-181a-5p and endoplasmic reticulum stress in the regulation of myogenic differentiation. Gene. 592:60–70. 2016.PubMed/NCBI View Article : Google Scholar
23	Li S, Yang J, Xia Y, Fan Q and Yang KP: Long noncoding RNA NEAT1 promotes proliferation and invasion via targeting miR-181a-5p in non-small cell lung cancer. Oncol Res. 26:289–296. 2018.PubMed/NCBI View Article : Google Scholar
24	Yu C, Sun J, Leng X and Yang J: Long noncoding RNA SNHG6 functions as a competing endogenous RNA by sponging miR-181a-5p to regulate E2F5 expression in colorectal cancer. Cancer Manag Res. 11:611–624. 2019.PubMed/NCBI View Article : Google Scholar
25	Mi Y, Zhang D, Jiang W, Weng J, Zhou C, Huang K, Tang H, Yu Y, Liu X, Cui W, et al: miR-181a-5p promotes the progression of gastric cancer via RASSF6-mediated MAPK signaling activation. Cancer Lett. 389:11–22. 2017.PubMed/NCBI View Article : Google Scholar
26	Zhang L, Pang S, Deng B, Qian L, Chen J, Zou J, Zheng J, Yang L, Zhang C, Chen X, et al: High glucose induces renal mesangial cell proliferation and fibronectin expression through JNK/NF-κB/NADPH oxidase/ROS pathway, which is inhibited by resveratrol. Int J Biochem Cell Biol. 44:629–638. 2012.PubMed/NCBI View Article : Google Scholar
27	Sodhi CP, Phadke SA, Batlle D and Sahai A: Hypoxia and high glucose cause exaggerated mesangial cell growth and collagen synthesis: Role of osteopontin. Am J Physiol Renal Physiol. 280:F667–F674. 2001.PubMed/NCBI View Article : Google Scholar
28	Suzaki Y, Yoshizumi M, Kagami S, Nishiyama A, Ozawa Y, Kyaw M, Izawa Y, Kanematsu Y, Tsuchiya K and Tamaki T: BMK1 is activated in glomeruli of diabetic rats and in mesangial cells by high glucose conditions. Kidney Int. 65:1749–1760. 2004.PubMed/NCBI View Article : Google Scholar
29	Singh L, Pushker N, Saini N, Sen S, Sharma A, Bakhshi S, Chawla B and Kashyap S: Expression of pro-apoptotic Bax and anti-apoptotic Bcl-2 proteins in human retinoblastoma. Clin Exp Ophthalmol. 43:259–267. 2015.PubMed/NCBI View Article : Google Scholar
30	Zhu Y, Tchkonia T, Fuhrmann-Stroissnigg H, Dai HM, Ling YY, Stout MB, Pirtskhalava T, Giorgadze N, Johnson KO, Giles CB, et al: Identification of a novel senolytic agent, navitoclax, targeting the Bcl-2 family of anti-apoptotic factors. Aging Cell. 15:428–435. 2016.PubMed/NCBI View Article : Google Scholar
31	Ghibelli L and Diederich M: Multistep and multitask Bax activation. Mitochondrion. 10:604–613. 2010.PubMed/NCBI View Article : Google Scholar
32	Caruso Bavisotto C, Nikolic D, Marino Gammazza A, Barone R, Lo Cascio F, Mocciaro E, Zummo G, Conway de Macario E, Macario AJ, Cappello F, et al: The dissociation of the Hsp60/pro-Caspase-3 complex by bis (pyridyl) oxadiazole copper complex (CubipyOXA) leads to cell death in NCI-H292 cancer cells. J Inorg Biochem. 170:8–16. 2017.PubMed/NCBI View Article : Google Scholar
33	Qi W, Holian J, Tan CY, Kelly DJ, Chen XM and Pollock CA: The roles of Kruppel-like factor 6 and peroxisome proliferator-activated receptor-γ in the regulation of macrophage inflammatory protein-3α at early onset of diabetics. Int J Biochem Cell Biol. 43:383–392. 2011.PubMed/NCBI View Article : Google Scholar
34	Ghahhari NM and Babashah S: Interplay between microRNAs and Wnt/β-catenin signaling pathway regulates epithelial-mesenchymal transition in cancer. Eur J Cancer. 51:1638–1649. 2015.PubMed/NCBI View Article : Google Scholar
35	Mao J, Fan S, Ma W, Fan P, Wang B, Zhang J, Wang H, Tang B, Zhang Q, Yu X, et al: Roles of Wnt/β-catenin signaling in the gastric cancer stem cells proliferation and salinomycin treatment. Cell Death Des. 5(e1039)2014.PubMed/NCBI View Article : Google Scholar
36	Gao L, Chen B, Li J, Yang F, Cen X, Liao Z and Long X: Wnt/β-catenin signaling pathway inhibits the proliferation and apoptosis of U87 glioma cells via different mechanisms. PLoS One. 12(e0181346)2017.PubMed/NCBI View Article : Google Scholar
37	Lan XQ, Wen HX, Aslam R, Marashi Shoshtari SS, Mishara A, Kumar V, Wang HC, Wu GS, Luo HR, Malhotra A and Singhal PC: Nicotine enhances mesangial cell proliferation and fibronectin production in high glucose milieu via activation of Wnt/β-catenin pathway. Bioscience Rep. 38: pii(BSR20180100)2018.PubMed/NCBI View Article : Google Scholar
38	Xu P, Guan MP, Bi JG, Wang D, Zheng ZJ and Xue YM: High glucose down-regulates microRNA-181a-5p to increase pro-fibrotic gene expression by targeting early growth response factor 1 in HK-2 cells. Cell Signal. 31:96–104. 2017.PubMed/NCBI View Article : Google Scholar