Long Non-Coding RNA Metastasis Associated Lung Adenocarcinoma Transcript 1 (MALAT1) Promotes Hypertension by Modulating the Hsa-miR-124-3p/Nuclear Receptor Subfamily 3, Group C, Member 2 (NR3C2) and Hsa-miR-135a-5p/NR3C2 Axis

Abstract

Background:This study was designed to investigate the role of long non-coding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) in the proliferation as well as apoptosis of human umbilical vein endothelial cells (HUVECs), to offer a basis for therapy of hypertension. Material/Methods:The lncRNA MALAT1 expression, hsa-miR-124-3p, hsa-miR-135a-5p, hsa-miR-135b-5p, and hsa-miR-455-5p in plasma were measured from 230 patients with hypertension and 230 non-hypertensive controls. The mechanism for lncRNA MALAT1 modulating the proliferation and apoptosis of HUVECs was explored by cell transfection, Cell Counting Kit-8 (CCK-8), quantitative real-time polymerase chain reaction (qRT-PCR), western blot, and dual-luciferase reporter assays. Results:The expression of hsa-miR-124-3p and hsa-miR-135a-5p was reduced and the expression of lncRNA MALAT1 was increased in the plasma of hypertensive patients. Moreover, the plasma levels of hsa-miR-124-3p and hsa-miR-135a-5p of hypertensive patients were negatively correlated with lncRNA MALAT1 (r=-0.64, -0.72; P<0.01, P<0.01, respectively). The level of nuclear receptor subfamily 3, group C, member 2 (NR3C2) protein was negatively correlated with hsa-miR-124-3p and hsa-miR-135a-5p (r=-0.74, -0.84; P<0.01, P<0.01, respectively). The proliferation of HUVECs was inhibited after the inhibition of MALAT. Additionally, after knocking down MALAT, the levels of hsa-miR-124-3p and hsa-miR-135a-5p in HUVECs were markedly increased, while the expression level of NR3C2 protein was decreased. The apoptotic rate of HUVECs after the transfection of MALAT1 small interfering RNA (si-MALAT1) (3.64±0.21%) was significantly reduced compared to that of transfected si-MALAT1 no template control (NC) (3.76±0.19%) and the control group (10.51±1.24%). Conclusions:LncRNA MALAT1 regulates proliferation and apoptosis of HUVECs through the hsa-miR-124-3p/NR3C2 and/or hsa-miR-135a-5p/NR3C2 axis.

LncRNA MALAT1-deficiency restrains lipopolysaccharide (LPS)-induced pyroptotic cell death and inflammation in HK-2 cells by releasing microRNA-135b-5p

“…Given that LncRNA MALAT1 positively regulates NLRP3-mediated cell pyroptosis [ 21 , 28 ], we hypothesized that some miRNAs might bridge LncRNA MALAT1 and NLRP3 mRNA. As expected, we screened out a novel miRNA-135b-5p that potentially targeted both LncRNA MALAT1 [ 37 ] and exerted its inhibiting effects on NLRP3 [ 38 , 39 ], and further results evidenced that LncRNA MALAT1 positively regulated NLRP3 expressions by sponging miRNA-135b-5p. Interestingly, miRNA-135b-5p reversed LPS induced inflammatory responses [ 40 ], and our data supported that LPS downregulated miRNA-135b-5p, and upregulation of miRNA-135b inhibited NLRP3-mediated cell pyroptosis to recover cellular functions in LPS-treated HK-2 cells.…”

“…Based on our preliminary work (data not shown), the HK-2 cells at passage 3–6 were subjected to LPS (2 μg/mL) treatment for 0, 24, and 48 h, respectively. In addition, the gene manipulating vectors for LncRNA MALAT1, and miRNA-135b-5p mimic and inhibitor were constructed by Sangon Biotech (Shanghai, China) according to the previous publications [ 37–39 ].…”

“…Human renal tubular epithelial HK-2 cells were bought from American Type Culture Collection (ATCC, Manassas, VA) and were maintained in the DMEM/F12 medium (Gibco, Waltham, MA) containing 10% FBS (Gibco, Waltham, MA) under the standard culture conditions with 5% CO 2 at 37 C. Based on our preliminary work (data not shown), the HK-2 cells at passage 3-6 were subjected to LPS (2 lg/mL) treatment for 0, 24, and 48 h, respectively. In addition, the gene manipulating vectors for LncRNA MALAT1, and miRNA-135b-5p mimic and inhibitor were constructed by Sangon Biotech (Shanghai, China) according to the previous publications [37][38][39].…”

“…Following the protocols provided by the previous studies [37][38][39], the real-time qPCR was performed to examine genes expression levels at transcriptional levels. Briefly, total RNA was obtained by TRIzol (Beyotime, Shanghai, China) and analyzed by the following agarose electrophoresis.…”

“…In addition, LncRNA MALAT1 exerts its inflammation-promoting effects through modulating miR-181c-5p [ 32 ], miR-26b [ 33 ], and miR-590 [ 34 ], and Dai et al notice that knock-down of LncRNA MALAT1 releases miR-146a to ameliorate LPS-induced acute lung injury [ 35 ], and LncRNA MALAT1 regulates LPS-induced cytokines secretions in human gingival fibroblasts by sponging miR-20a [ 36 ]. Among all the miRNAs, miRNA-135b-5p targets both LncRNA MALAT1 [ 37 ] and 3′ untranslated regions of NLRP3 mRNA [ 38 , 39 ], and Li et al evidence that miRNA-135b-5p silences AMPK phosphatase Ppm1e to restrain LPS-mediated production of pro-inflammatory cytokines [ 40 ]. In addition, miRNA-135b-5p is reported to be relevant to renal fibrosis [ 41 ].…”