Akt2 inhibitor promotes M2 macrophage polarization in rats with periapical inflammation by reducing miR-155-5p expression / 南方医科大学学报

OBJECTIVE@#To investigate the effect of Akt2 inhibitor on macrophage polarization in the periapical tissue in a rat model of periapical inflammation.@*METHODS@#Rat models of periapical inflammation were established in 28 normal SD rats by opening the pulp cavity of the mandibular first molars, followed by injection of normal saline and Akt2 inhibitor into the left and right medullary cavities, respectively. Four rats without any treatment served as the healthy control group. At 7, 14, 21 and 28 days after modeling, 7 rat models and 1 control rat were randomly selected for observation of inflammatory infiltration in the periapical tissues by X-ray and HE staining. Immunohistochemistry was used to detect the expression and localization of Akt2, macrophages and the inflammatory mediators. RT-PCR was performed to detect the mRNA expressions of Akt2, CD86, CD163, inflammatory mediators, miR-155-5p and C/EBPβ to analyze the changes in macrophage polarization.@*RESULTS@#X-ray and HE staining showed that periapical inflammation was the most obvious at 21 days after modeling in the rats. Immunohistochemistry and RT-PCR showed that compared with those in the control rats, the expressions of Akt2, CD86, CD163, miR-155-5p, C/EBPβ, and IL-10 increased significantly in the rat models at 21 days (P < 0.05). Compared with saline treatment, treatment with the Akt2 inhibitor significantly decreased the expression levels of Akt2, CD86, miR-155-5p and IL-6 and the ratio of CD86+M1/CD163+M2 macrophages (P < 0.05) and increased the expression levels of CD163, C/EBPβ and IL-10 in the rat models (P < 0.05).@*CONCLUSION@#Inhibition of Akt2 can delay the progression of periapical inflammation in rats and promote M2 macrophage polarization in the periapical inflammatory microenvironment possibly by reducing miR-155-5p expression and activating the expression of C/EBPβ in the Akt signaling pathway.

Testicular exosomes disturb the immunosuppressive phenotype of testicular macrophages mediated by miR-155-5p in uropathogenic Escherichia coli-induced orchitis / 亚洲男科学杂志(英文版)

Male reproductive infections are known to shape the immunological homeostasis of the testes, leading to male infertility. However, the specific pathogenesis of these changes remains poorly understood. Exosomes released in the inflammatory microenvironment are important in communication between the local microenvironment and recipient cells. Here, we aim to identify the immunomodulatory properties of inflammatory testes-derived exosomes (IT-exos) and explore their underlying mechanisms in orchitis. IT-exos were isolated using a uropathogenic Escherichia coli (UPEC)-induced orchitis model and confirmed that IT-exos promoted proinflammatory M1 activation with increasing expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in vitro. We further used small RNA sequencing to identify the differential miRNA profiles in exosomes and primary testicular macrophages (TMs) from normal and UPEC-infected testes, respectively, and identified that miR-155-5p was highly enriched in IT-exos and TMs from inflammatory testes. Further study of bone marrow derived macrophages (BMDMs) transfected with miR-155-5p mimic showed that macrophages polarized to proinflammatory phenotype. In addition, the mice that were administrated IT-exos showed remarkable activation of TM1-like macrophages; however, IT-exos with silencing miR-155-5p showed a decrease in proinflammatory responses. Overall, we demonstrate that miR-155-5p delivered by IT-exos plays an important role in the activation of TM1 in UPEC-induced orchitis. Our study provides a new perspective on the immunological mechanisms underlying inflammation-related male infertility.

miR-155-5p Expression, Function and Regulation in Tumors / 肿瘤防治研究

MicroRNAs (miRNAs) are a class of small, single-stranded non-coding RNAs that act as important regulators of gene expression and are involved in a number of important processes in life. A large number of studies have suggested that dysregulation of miRNA expression may be an important part of the mechanism of human tumorigenesis and progression. MiR-155-5p is mainly regarded as an oncomiR that acts on multiple target genes to participate in tumor progression, although it has been suggested to possess cancer growth suppressor effects. In this paper, we summarize the effects of miR-155-5p on cancer cell proliferation, invasion, migration, and drug resistance in various tumor types and elucidate its value as a possible potential marker in assisting diagnosis.

Expression of miR-155-5p in Wilms tumor and its regulatory role in proliferation, migration and apoptosis of Wilms tumor cells / 南方医科大学学报

OBJECTIVE@#explore the expression of miR-155-5p in Wilms tumor and its effect in regulating the proliferation, migration and apoptosis of Wilms tumor cells.@*METHODS@#Specimens of tumor tissues and paired adjacent tissues were obtained from 40 patients with Wilms tumor for detection of the expression levels of miR-155-5p using RT-qPCR. Wilms tumor cell line G401 was transfected with miR-155-5p mimics and miR-155-5p inhibitor to induce miR-155-5p over-expression and its inhibition, respectively, and the changes in the cell proliferation, migration and apoptosis were assessed using cell counting kit-8 (CCK-8), wound healing assay and fl ow cytometry.@*RESULTS@#RT-qPCR showed that the expression of miR-155-5p decreased significantly in Wilms tumor tissues as compared with normal kidney tissues and was significantly associated with TNM stage ( < 0.05). In G401 cells, over-expression of miR-155-5p significantly inhibited the cell proliferation and migration and promoted cell apoptosis ( < 0.05), and down-regulation of miR-155-5p obviously enhanced the proliferation and migration and suppressed apoptosis of the cells ( < 0.05).@*CONCLUSIONS@#miR-155-5p is down-regulated in Wilms tumor and its expression level is correlated with TNM stage. miR-155-5p participates in the progression of Wilms tumor by inhibiting the proliferation and migration and promoting apoptosis of the tumor cells, and may serve as a novel biomarker for diagnosis, therapy and prognostic evaluation of Wilms tumor.

XIST Induced by JPX Suppresses Hepatocellular Carcinoma by Sponging miR-155-5p

PURPOSE: The influence of X-inactive specific transcript (XIST) and X-chromosome inactivation associated long non-coding RNAs (lncRNAs) just proximal to XIST (JPX) on hepatocellular carcinoma (HCC) remains controversial in light of previous reports, which the present study aimed to verify. MATERIALS AND METHODS: The DIANA lncRNA-microRNA (miRNA) interaction database was used to explore miRNA interactions with JPX or XIST. JPX, XIST, and miR-155-5p expression levels in paired HCC specimens and adjacent normal tissue were analyzed by RT-qPCR. Interaction between XIST and miR-155-5p was verified by dual luciferase reporter assay. Expression levels of miR-155-5p and its known target genes, SOX6 and PTEN, were verified by RT-qPCR and Western blot in HepG2 cells with or without XIST knock-in. The potential suppressive role of XIST and JPX on HCC was verified by cell functional assays and tumor formation assay using a xenograft model. RESULTS: JPX and XIST expression was significantly decreased in HCC pathologic specimens, compared to adjacent tissue, which correlated with HCC progression and increased miR-155-5p expression. Dual luciferase reporter assay revealed XIST as a direct target of miR-155-5p. XIST knock-in significantly reduced miR-155-5p expression level and increased that of SOX6 and PTEN, while significantly inhibiting HepG2 cell growth in vitro, which was partially reversed by miR-155-5p mimic transfection. JPX knock-in significantly increased XIST expression and inhibited HepG2 cell growth in vitro or tumor formation in vivo in a XIST dependent manner. CONCLUSION: JPX and XIST play a suppressive role in HCC. JPX increases expression levels of XIST in HCC cells, which suppresses HCC development by sponging the cancer promoting miR-155-5p.

miR-155-5p Promotes Invasion and Migration of MCF-7 Breast Cancer Cell by Down-regulating SOX1 / 华中科技大学学报(医学版)

Objective To examine the mechanism by which miR-155-5p regulates sex determining region Y-boc 1(SOX1) expression in breast cancer cell line MCF-7,and to investigate the effect of miR-155-5p on the invasion and metastasis of these cells.Methods After up-regulation or down-regulation of the expression of miR-155-5p in MCF-7 cells,the transwell assay was performed to detect the invasive and migrating ability of MCF-7 cells;SOX1 mRNA expression and protein levels were detected by real-time PCR and Western blot,respectively;immunofluorescence was used to test the distribution and levels of SOX1 pro-tein,and dual luciferase reporter experiments to discover the binding of miR-155-5p to the 3′-UTR region of SOX1 mR-NA.Results Up-regulating the expression of miR-155-5p could enhance the migrating and invasive ability of MCF-7 cells and simultaneously decrease the mRNA and protein levels of SOX1(P <0.05).On the contrary,inhibition of miR-155-5p depressed the invasion and migration of MCF-7 cells and increased the expression of SOX1(P < 0.05).Dual luciferase reporter experi-ments confirmed that miR-155-5p could bind to the specific sequence in the 3′-UTR of SOX1 mRNA and play a role of regula-tor.Conclusion miR-155-5p promotes the invasion and migration of breast cancer cell line MCF-7 via inhibiting the expression of SOX1.

Expression and roles of miR-155-5p in esophageal squamous cell carcinoma / 医学研究生学报

Objective Studies show that miRNA plays an important role in regulating the development, metastasis, inva-sion, and angiogenesis of carcinoma.This study aimed to investigate the expression of miR-155-5p in esophageal squamous cell carci-noma ( ESCC) and its role in the occurrence and progression of ESCC. Methods We collected 20 surgical specimens of ESCC and another 20 from the adjacent normal tissue at least 5 cm from the lesion and determined the expression of miR-155-5p in the tissues by qPCR.We transfected the miR-155-5p mimic, inhibitor and negative control plasmid into the Eca109 cells, measured the proliferation and apoptosis of the cells by CCK8 assay and flow cytometry, and examined their migration using the scratch test. Results The ex-pression of miR-155-5p was significantly up-regulated in the ESCC tissue as compared with that in the normal tissue (529.42% vs 100%, P<0.05).The scratch test showed a markedly increased migration of the cancer cells when miR-155-5p was overexpressed. No remarkable differences were observed in the apoptosis rates of the cells transfected with the miR-155-5p mimic ([5.43 ± 3.09]%), inhibitor ([5.28 ±1.98]%), and negative control plasmid ([5.67 ±1.99]%) (P<0.05). Conclusion The ex-pression of miR-155-5p is significantly up-regulated in the ESCC tissue, which may act as a potential maker for the diagnosis of ESCC. The up-regulated expression of miR-155-5p does not significantly in-fluence the proliferation and apoptosis but enhances the migration of Eca109 cells, which may be related to early metastasis of ESCC.

Study of miR-125b, miR-29a and miR-155-5p as biomarkers of tuberculous meningitis / 天津医药

Objective To investigate the expression levels of microRNA(miR)-125b, miR-29a and miR-155-5p in ce?rebrospinal fluid (CSF) and plasma from the patients with tuberculous meningitis and their clinic significance. Methods Cerebrospinal fluid and plasma samples were collected from 20 patients with tuberculous meningitis (tuberculous meningitis group) and 20 patients suffered from primary headache (control group). The total RNAs were extracted.The levels of miR-125b, miR-29a and miR-155-5p were determined by real-time quantitative PCR (q-PCR). Results Levels of miR-29a and miR-125b in both CSF and plasma of patients in tuberculous meningitis group were significantly higher than those in pa?tients from control group with statistical significance (P<0.01). Mean time, the level of miR-155-5p in plasma but not in CSF of patients in tuberculous meningitis group was higher than those in control group with statistical significance ( P<0.01). Conclusion Expression of miR-125b, miR-29a and miR-155-5p may participate in regulating the occurrence and development of tuberculous meningitis. And miR-125b and miR-29a may be used as potential biomarkers for diagnosing tu?berculous meningitis.