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Long non-cording RNA XIST promoted cell proliferation and suppressed apoptosis by miR-423-5p/HMGA2 axis in diabetic nephropathy

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Abstract

This research studied the effect of long non-coding RNA X-inactive-specific transcript (XIST) on DN. The effect of high glucose (HG) on the expression of XIST and miR-423-5p was detected by quantitative real-time PCR (qRT-PCR) in human kidney (HK) cells (human glomerular mesangial cells (HMCs) and human kidney-2 (HK-2) cells). The effect of XIST depletion and miR-423-5p inhibition or overexpression on high mobility group protein A2 (HMGA2) protein level was examined by western blot in HG-induced HK cells. The impacts of XIST depletion on viability and apoptosis were assessed by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) and flow cytometry assays in HG-induced HK cells. We found the expression of XIST and HMGA2 protein was significantly upregulated in DN tissues and cells. Moreover, HG treatment induced the upregulation of XIST and HMGA2 protein level in HK cells. Besides, both XIST depletion and HMGA2 depletion decreased cell proliferation but increased apoptosis in HG-treated HK cells. Furthermore, HMGA2 upregulation or miR-423-5p inhibition partly eliminated the effects of XIST depletion on cell proliferation, apoptosis of HG-treated HK cells. Interestingly, HMGA2 upregulation partly reversed miR-423-5p overexpression-mediated suppression on viability and promotion on apoptosis in HG-treated HK cells. Mechanistically, XIST sponged miR-423-5p to regulate HMGA2 expression in DN cells. Taken together, XIST depletion suppressed proliferation and promoted apoptosis via miR-423-5p/HMGA2 axis in HG-treated HK cells, which may provide a potential therapeutic target for DN.

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Data availability

The analyzed data sets generated during the present study are available from the corresponding author on reasonable request.

Abbreviations

DN:

Diabetic nephropathy

CKD:

Chronic kidney diseases

ESKD:

End-stage kidney disease

ECM:

Extracellular matrix

XIST:

Long non-coding RNA X-inactive-specific transcript

HMCs:

Human glomerular mesangial cells

qRT-PCR:

RNA isolation and quantitative real-time polymerase chain reaction

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Funding

This research was funded by China Health Promotion Foundation No. hxkt2018-8 and Basic research program of Jiangsu Province (Natural Science Foundation)—General research Projects No. BK2016323.

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Authors and Affiliations

  1. Department of Endocrinology and Metabolism, The First Affiliated Hospital of Soochow University, 188 Shizi Road, Suzhou, 215006, Jiangsu, China

    Hui Chen & Xingbo Cheng

  2. Department of Endocrinology and Metabolism, Northern Jiangsu People’s Hospital, Clinical Medical College of Yangzhou University, Yangzhou, Jiangsu, China

    Hui Chen

  3. Clinical Laboratory Center, Northern Jiangsu People’s Hospital, Yangzhou, Jiangsu, China

    Yuan Guo

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  1. Hui Chen
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  2. Yuan Guo
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All authors made substantial contribution to conception and design, acquisition of the data, or analysis and interpretation of the data; take part in drafting the article or revising it critically for important intellectual content; gave final approval of the revision to be published; and agree to be accountable for all aspects of the work.

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Correspondence to Xingbo Cheng.

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The present study was approved by the ethical review committee of The First Affiliated Hospital of Soochow University. Written informed consent was obtained from all enrolled patients.

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11010_2021_4250_MOESM1_ESM.tif

Supplementary file1 Fig. S1 The expression level of HMGA2 in DN tissues and normal tissues. IHC staining of HMGA2 protein in human DN tissues and normal tissues (brown staining indicated positive signal) (TIF 1749 KB)

11010_2021_4250_MOESM2_ESM.tif

Supplementary file2 Fig. S2 The effects of si-XIST or si-HMGA2 on XIST, HMGA2 and miRNAs expression. (A) Relative expression of XIST in HMCs and HK-2 cells transfected with si-NC, si-XIST-1, si-XIST-2 or si-XIST-3, respectively. (B) Relative expression of HMGA2 in HMCs and HK-2 cells transfected with si-NC, si-HMGA2-1, si-HMGA2-2 or si-HMGA2-3, respectively. (C,D) The expression levels of miR-362-3p, miR-142-5p, miR-455-3p, miR-27a-3p and miR-423-5p in HMCs and HK-2 cells transfected with si-NC or si-XIST. * P<0.05 (TIF 660 KB)

11010_2021_4250_MOESM3_ESM.tif

Supplementary file3 Fig. S3 The effect of XIST overexpression plasmids on HMGA2 expression. Cells were transfected with vector or XIST overexpression plasmids. And the expression of HMGA2 mRNA in HG-treated HK cells was detected. * P<0.05 (TIF 167 KB)

11010_2021_4250_MOESM4_ESM.tif

Supplementary file4 Fig. S4 The effect of miR-423-5p inhibitor on fibrosis in HG-treated HK cells. (A, B) Cells were transfected with inhibitor NC or miR-423-5p inhibitor. And the mRNA levels of fibronectin (FN) and collagen IV (CoII IV) in HG-treated HK cells were detected. * P<0.05 (TIF 280 KB)

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Chen, H., Guo, Y. & Cheng, X. Long non-cording RNA XIST promoted cell proliferation and suppressed apoptosis by miR-423-5p/HMGA2 axis in diabetic nephropathy. Mol Cell Biochem 476, 4517–4528 (2021). https://doi.org/10.1007/s11010-021-04250-x

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