Long Non-coding RNA MEG3 Promotes Pyroptosis in Testicular Ischemia-Reperfusion Injury by Targeting MiR-29a to Modulate PTEN Expression

Abstract

Increasing evidence shows that the abnormal long non-coding RNAs (lncRNAs) expression is closely related to ischemia-reperfusion injury (I/R) progression. Studies have previously described that lncRNA MEG3 regulates pyroptosis in various organs I/R. Nevertheless, the related mechanisms of MEG3 in testicular I/R has not been clarified. The aim of this research is to unravel underlying mechanisms of the regulation of pyroptosis mediated by MEG3 during testicular I/R. We have established a testicular torsion/detorsion (T/D) model and an oxygen-glucose deprivation/reperfusion (OGD/R)-treated spermatogenic cell model. Testicular ischemic injury was assessed by H&E staining. Western blotting, quantitative real-time PCR, MDA, and SOD tests and immunohistochemistry measured the expression of MEG3 and related proteins and the level of ROS production in testicular tissues. Quantitative real-time PCR and western blotting determined the relative expression of MEG3, miR-29a, and relevant proteins in GC-1. Cell viability and cytotoxicity were measured by CCK-8 and LDH assays. Secretion and expression levels of inflammatory proteins were determined by ELISA, immunofluorescence and western blotting. The interaction among MEG3, miR-29a, and PTEN was validated through a dual luciferase reporter assay and Ago2-RIP. In this research, we identified that MEG3 was upregulated in animal specimens and GC-1. In loss of function or gain of function assays, we verified that MEG3 could promote pyroptosis. Furthermore, we found that MEG3 negatively regulated miR-29a expression at the posttranscriptional level and promoted PTEN expression, and further promoted pyroptosis. Therefore, we explored the interaction among MEG3, miR-29a and PTEN and found that MEG3 directly targeted miR-29a, and miR-29a targeted PTEN. Overexpression of miR-29a effectively eliminated the upregulation of PTEN induced by MEG3, indicating that MEG3 regulates PTEN expression by targeting miR-29a. In summary, our research indicates that MEG3 contributes to pyroptosis by regulating miR-29a and PTEN during testicular I/R, indicating that MEG3 may be a potential therapeutic target in testicular torsion.

The protective effect of p‐Coumaric acid on hepatic injury caused by particulate matter in the rat and determining the role of long noncoding RNAs MEG3 and HOTAIR

“…Increased expression of MEG3 inhibits the progression of hepatic ischemia‐reperfusion injury. [ 7 ] Homeobox protein antisense intergenic RNA (HOTAIR) is also a LncRNA that increased expression of it in hepatocellular carcinoma patients who have undergone hepatectomy or liver transplantation causes recur of cancer shortly after surgery. It has also been shown that HOTAIR levels increase due to hepatic I/R damage.…”

Single-cell transcriptome profiling reveals several LncRNAs differentially expressed in idiopathic germ cell aplasia

“…MEG3, overexpressed in LEY, MYD and STRO cell types of our patients, was an interesting molecule also because of its role as a maternally imprinted transcript ( Schmidt et al, 2000 ). Moreover, given the relevant role played by inflammation in the testes of our patients ( Alfano et al, 2021 ), it was i intriguing that, recently, MEG3 was found to promote pyroptosis, i.e., a form of cell-death associated with inflammatory signals, in testicular ischemia-reperfusion (IR) injury ( Ning et al, 2021 ). In order to evaluate the consistency of the MEG3 role in IR injury, we performed a literature search using pertinent keywords (see Methods).…”

A Novel Risk Model Identified Based on Pyroptosis-Related lncRNA Predicts Overall Survival and Associates With the Immune Landscape of GC Patients

“…In addition, a large number of lncRNAs were identified as prognostic biomarkers in multiple cancers (Jin et al, 2020;Shen et al, 2020;Wu et al, 2020). In recent years, lncRNAs were widely reported to modulate pyroptosis in many diseases including cancer (Chen et al, 2021;Huang et al, 2021;Ning et al, 2021;Wang. L.-Q et al, 2021).…”

“…W.-X et al, 2017;Chi et al, 2019). Furthermore, lncRNAs were widely reported to modulate pyroptosis in many diseases including cancer (Chen et al, 2021;Huang et al, 2021;Ning et al, 2021;Wang. L.-Q et al, 2021).…”