MicroRNA-128b mediates lipopolysaccharide-induced apoptosis via reactive oxygen species in human pulmonary microvascular endothelial cells

ABSTRACT Objectives: This study aimed to explore the effects of miR-128b in the regulation of Lipopolysaccharide (LPS) induced apoptosis. Methods: Human Pulmonary Microvascular Endothelial Cells (HPMECs) were transfected with an miR-128b inhibitor and stimulated with LPS for 24 h. FCM was performed to detect apoptosis and Reactive Oxygen Species (ROS) production. In addition, miRNA and caspase-3 expression levels were determined using real-time quantitative polymerase chain reaction and western blotting. Results: LPS significantly induced apoptosis and ROS production and upregulated miR-128b and caspase-3 expressions in HPMECs. However, LPS-induced effects were suppressed when an miR-128b inhibitor was used. Preincu-bation with NAC decreased the LPS-induced apoptosis of HPMECs. Conclusions: These effects were mediated by miR-128b via the caspase-3 pathway.

SNHG22 promotes migration and invasion of trophoblasts via miR-128-3p/ PCDH11X axis and activates PI3K/Akt signaling pathway

Abstract Objectives Long non-coding RNAs (LncRNAs) act as an indispensable role in the Preeclampsia (PE)-related trophoblast function, while its relationship with Small Nucleolar RNA Host Gene 22 (SNHG22) remains unknown. Hence, this study aimed to investigate the roles of lncRNA SNHG22 in the Preeclampsia (PE)-related trophoblasts function and the underlying mechanism. Methods Normal placentas and placentas from PE patients were collected to detect the expression of lncRNA SNHG22. Then, trophoblasts HTR-8/Svneo and JEG-3 were purchased, cultured, and treated to investigate the roles of lncRNA SNHG22 on cell migration and invasion as well as its underlying regulatory mechanism. Results The SNHG22 was downregulated in PE patients, and it was found that SNHG22 overexpression could drive migration and invasion of trophoblasts, while SNHG22 depletion exerted a suppressive effect. Mechanistically, SNHG22 was validated to regulate microRNA-128-3p (miR-128-3p), and Protocadherin 11 X-Linked (PCDH11X) was identified as the target gene of miR-128-3p. Furthermore, it was found that SNHG22 acted as a promoter in the migration and invasion of trophoblast cells in a miR-128-3p/PCDH11X dependent manner, and SNHG22 silencing weakened the activation of PCDH11X-mediated PI3K/Akt signaling pathways through inhibiting miR-128-3p, thereby preventing migration and invasion of trophoblasts. Conclusion SNHG22 acted as a driver in the migration and invasion of trophoblasts and may be considered a candidate for the amelioration of PE.

MiR-128-3p Regulates Proliferation, Migration and Apoptosis of Glioblastoma Multiforme by Targeting HOXA5 / 肿瘤防治研究

Objective To investigate the reasons of HOXA5 overexpression in GBM and the molecular mechanism of miR-128-3p regulating the proliferation, invasion and apoptosis of glioblastoma multiforme. Methods After increasing and decreasing miR-128-3p expression in U87 cell lines by lentivirus transfection, the changes of HOXA5 expression were detected by Western blot, to explore the correlation between miR-128-3p and HOXA5 in GBM. The dual-luciferase reporter tests were performed to detect the target interaction of miR-128-3p with HOXA5. Through CCK-8 test, Transwell test, flow cytometric assay and tumor cell xenograft in nude mice, we verified molecular mechanism of miR-128-3p regulating the proliferation, invasion and apoptosis of GBM in vitro and in vivo. Results The expression level of HOXA5 was decreased in U87 cell line after miR-128-3p upregulation. In addition, the expression level of HOXA5 was increased in U87 cell line after miR-128-3p downregulation (P < 0.05). The expression level of HOXA5 was correlated negatively with the expression of miR-128-3p in U87 cell lines. MiR-128-3p targetedly interacted with 3'UTR of HOXA5 and inhibited the expression of HOXA5. The proliferation, invasion and anti-apoptosis of U87 cells were significantly decreased in the miR-128-3p+control group. Conclusion MiR-128-3p regulates negatively the proliferation, invasion and anti-apoptosis of GBM cells by targeting HOXA5. The overexpression of HOXA5 is induced by downregulation of miR-128-3p in GBM.

Diagnostic and prognostic value of serum miR-9-5p and miR-128-3p levels in early-stage acute ischemic stroke

OBJECTIVES: To investigate the clinical utility of serum microRNA levels (miR-9-5p and miR-128-3p) in the diagnosis and prognosis of early-stage acute ischemic stroke (AIS). METHODS: We compared the differences in serum miR-9-5p and miR-128-3p levels between patients with AIS and healthy individuals (controls). The serum levels of miR-9-5p and miR-128-3p were quantified using quantitative real-time PCR, and the association of each miRNA with AIS was determined using receiver operator characteristic curve analysis. The predictive value of these indices in the diagnosis of early-stage AIS was evaluated in conjunction with that of computed tomography findings and neuron-specific enolase levels. The prognosis of patients with AIS was evaluated three months after their discharge from hospital using the modified Rankin scale, which classifies the prognosis as either favorable or poor. Logistic regression analysis was used to analyze the correlation between miR-9-5p and miR-128-3p levels and patient prognosis. RESULTS: The serum levels of miR-9-5p and miR-128-3p were upregulated in patients with AIS relative to those in healthy individuals. A pronounced correlation was identified between serum miR-9-5p and miR-128-3p levels and patient prognosis, with high levels of both miRNAs being associated with poor patient outcomes. CONCLUSION: Assessment of serum miR-9-5p and miR-128-3p levels is important for the early diagnosis and prognosis of AIS.

MiR-128 suppresses metastatic capacity by targeting metadherin in breast cancer cells

BACKGROUND: Breast cancer, the most common cancer in women worldwide, causes the vast majority of cancer-related deaths. Undoubtedly, tumor metastasis and recurrence are responsible for more than 90 percent of these deaths. MicroRNAs are endogenous noncoding RNAs that have been integrated into almost all the physiological and pathological processes, including metastasis. In the present study, the role of miR-128 in breast cancer was investigated. RESULTS: Compared to the corresponding adjacent normal tissue, the expression of miR-128 was significantly suppressed in human breast cancer specimens. More importantly, its expression level was reversely correlated to histological grade of the cancer. Ectopic expression of miR-128 in the aggressive breast cancer cell line MDA-MB-231 could inhibit cell motility and invasive capacity remarkably. Afterwards, Metadherin (MTDH), also known as AEG-1 (Astrocyte Elevated Gene 1) and Lyric that implicated in various aspects of cancer progression and metastasis, was further identified as a direct target gene of miR-128 and its expression level was up-regulated in clinical samples as expected. Moreover, knockdown of MTDH in MDA-MB-231 cells obviously impaired the migration and invasion capabilities, whereas re-expression of MTDH abrogated the suppressive effect caused by miR-128. CONCLUSIONS: Overall, these findings demonstrate that miR-128 could serve as a novel biomarker for breast cancer metastasis and a potent target for treatment in the future.

Expression of lncRNA SNHG16 in colorectal cancer tissues and cells and its regulationon expression of GPAM in colon cancer cells / 中国肿瘤生物治疗杂志

@# Objective: To investigate the expression of long non-coding RNASNHG16 (lncRNASNHG16) in colorectal cancer (CRC) tissues and cells, and to explore the mechanism of its regulation on the expression of mitochondrial glycerol-3-phosphate acyltransferase (GPAM) via sponging miR-128-3p. Methods: Sixty pairs of colorectal cancerous tissues and para-cancerous tissues that resected from CRC patients, who underwent surgery in the Department of Anorectal Surgery, Gansu Provincial People’s Hospital during Jan. 2014 and Jan. 2017, were collected for this study; In addition, CRC cell lines (SW480, SW620, HCT116, Caco-2,DLD-1, HT29) and colonic epithelial cell line CCD841 were also collected for the study. The expression of SNHG16 in collected tissues and cell lines was determined by Real-time quantitative PCR (qPCR), and its correlation to the clinicopathological features of CRC patients was also analyzed. SW480 cells were transfected with miR-128-3p mimic, miR-128-3p inhibitor, and si-SNHG16, respectively, and then the mRNA expressions of miR-128-3p and SNHG16 were detected by qPCR, the protein expression of GPAM was determined by Western blotting, and the cell proliferation, apoptosis and invasion were detected by CCK-8 assay, colony formation assay, cell apoptosis assay and Transwell chamber assay, respectively. The binding between SNHG16 and miR-128-3p was validated with dual luciferase reporter gene assay and RNA Immunoprecipitation assay. For in vivo experiment, mouse model of SW480 cell exnograft was constructed, and the effect of SNHG16 knockdown on the growth of exnograft was observed. Results: SNHG16 was found to highly expressed in human CRC tissues and cell lines (all P<0.01), and SNHG16 expression level was associated with lymph node metastasis, Duke's stage and patients’survival (all P<0.01). Knockdown of SNHG16 significantly inhibited CRC cell proliferation and invasion, and induced apoptosis (all P<0.01); After SNHG16 knockdown, the volume of exnograft was obviously reduced (P<0.05). Dual luciferase reporter gene assay and RNA Immunoprecipitation assay validated the interaction between miR-128-3p and SNHG16, and they were negatively correlated with each other in CRC patients (P<0.01). The SNHG16 regulated the expression of its down-stream gene GPAM via endogenously sponging miR-128-3p. Conclusion: SNHG16 regulates GPAM expression in CRC cells by sponging miR-128-3p, and SNHG16 and miR-128-3p may serve as potential targets for the diagnosis and treatment of CRC.

Determination of serum miR-128 level in patients with first-visit Parkinson’s disease and its related study / 临床神经病学杂志

To study the changes of serum miR-128 in patients with first-visit Parkinson’s disease (PD) before and after treatment, and the correlation between serum miR-128 and unified PD rating scale (UPDRS) scores and inflammatory factors in patients with PD. It is helpful to explore the diagnostic value and pathogenesis of miR-128 in PD. Methods Serum miR-128 levels were measured in 54 patients with first-visit PD ( first-visit PD group) before and after treatment, and were compared with 50 cases of healthy controls (healthy control group). The UPDRS scale was evaluated and the serum levels of IL-1β and TNF-α were measured, and the results were analyzed. Results The level of serum miR-128 in first-visit PD group was significantly lower than that in healthy control group (t = 8. 87, P< 0.01 ). After two months of treatment, the level of serum miR-128 in first-visit PD group was significantly higher than that before treatment (t= -5.13, P<0.01), and the UPDRS score was significantly lower than that before treatment (t=9.67, P<0.01). There was a negative correlation between serum miR-128 level and UPDRS score, IL-1β and TNF-α levels in first-visit PD group, respectively ( r= -0.763, r= -0.656, r= -0.674; all P<0.01). The area under the working characteristic curve of serum miR128 was 0.882 (95% CI:0.776-0. 952, P<0.01 ). The sensitivity of diagnosis of PD was 72. 0% , and the specificity was 88. 9%. Conclusion Among the first first-visit PD patients, the level of serum miR-128 is abnormal, and it can be used as a better index of peripheral blood for evaluating the condition and auxiliary diagnosis of PD, which plays an important role in the pathogenesis of depression.

miR-128 affects biological behavior of breast cancer cells through the targeting of RECK / 局解手术学杂志

Objective To investigate the biological behavior of breast cancer cells induced by miR-128 targeting RECK and its possible mechanism.Methods The expression of miR-128 in cell lines and transfection efficiency of lentiviral were detected by Western blot.Flow cytometry was used to detect the effect of miR-128 on the apoptosis of breast cancer cells.The dual luciferase assay was used to detect the in-teraction of miR-128 and RECK in breast cancer cells.Flow cytometry was used to detect the reversal effect of RECK on miR-128 in the cell cycle and colony formation of breast cancer cells.Results The expression of miR-128 in MCF-7 cells was 3.05 times that of NC cells,and the lentiviral transfection efficiency was obvious.miR-128 inhibits the growth of MCF-7 cells by inducing apoptosis and inhibiting cell cycle progression.RECK was the direct target of miR-128,and miR-128 directly regulated the expression of RECK.RECK ectopic recovery,miR-128 on breast cancer cell apoptosis and cell cycle inhibition had also been partially restored.Conclusion miR-128 plays an important role in the development and progression of breast cancer.It can interact with RECK to regulate the biological behavior of breast cancer cells,which suggests that miR-128 and RECK may be the potential therapeutic targets of breast cancer.

Effect of miR-128 and AK2 on biological behavior of cervical cancer cells by STAT3 signaling pathway / 中国免疫学杂志

Objective:To investigate the mechanism of miR-128 on the expression of AK2 protein through the STAT3 signaling pathway on the biological behavior of cervical cancer cells .Methods: The expression of AK2 and miR-128 in cervical cancer tissues and cells was detected by qPCR and Western blot .Double luciferase assay was used to detect the interaction between miR-128 and AK2.CCK-8 proliferation assay was used to detect the effect of miR-128 on the enhancement of cervical cancer cells .The effect of miR-128 on the tumorigenesis of cervical cancer cells was exa mined by tumor formation test in nude mice .Western blot was used to detect the effect of miR-128 on STAT3 signal pathway protein level .Western blot was used to detect the inhibitory effect of miR-128 on p-STAT3 by overexpressing AK2 protein.Results:The expression level of AK2 in cervical cancer tissues was higher than that in normal cervical tissues,and the expression level of miR-128 in cervical cancer C33a cells was lower.Double luciferase assay confirmed that miR-128 could directly target the expression of AK 2.CCK-8 proliferation test showed that miR-128 could inhibit the proliferation of cervical cancer cell lines .In vivo tumorigenesis test showed that the increase of miR-128 could inhibit the tumorigenesis ability of cervical cancer cells[volume(3.05±0.35)cm3 vs (0.86±0.11)cm3,P=0.031;weight(3.26±0.39)g vs (0.89±0.15)g,P=0.016 ];Western blot showed that miR-128 could inhibit the activation of p-STAT [ ( 42.12 ±6.28 )% vs ( 91.25 ±9.29 )%, P<0.05 ] ,while the overexpression of AK 2 could reverse the inhibitory effect of miR-128 on p-STAT.Conclusion: miR-128 is used to regulate the expression of AK 2 and regulate the biological behavior of cervical cancer cells through the activation of STAT 3 pathway.

MiR-128b is down-regulated in gastric cancer and negatively regulates tumour cell viability by targeting PDK1/Akt/NF-κ B axis.

Gastric cancer (GC) is the fourth most prevalent type of cancer worldwide, which is usually caused by the interaction between environmental and genetic factors, or epigenetic aspects. Referring to the non-coding RNAs, miR-128b has been reported to be associated with many tumour cases, and exerts distinct functions in different types of cancers. However, the function of miR-128b in GC onset and progression largely remains unknown. In the present study, we found that miR-128b expression was down-regulated in tissues from 18 GC patients and 3 carcinoma cell lines. In turn, over-expression of miR-128b suppressed GC cell proliferation, invasion and promoted apoptosis. Moreover, miR-128b was predicted to bind the 3 UTR of PDK1 gene using bioinformatic target-screening tools. Accordingly, ectopic expression of miR-128b inhibited the PDK1 expression at both transcriptional and post-transcriptional levels, and furthermore, the expression of gene tailed by the 3 UTR of PDK1 gene was significantly decreased in a dualluciferase reporter assay, suggesting that PDK1 was a direct target of miR-128b in GC cells. In the conditon of miR- 128b over-expression, we also observed spontaneous inactivation of the Akt/NF-κB signalling, implying PDK1 was a potential regulator of this pathway. In conclusion, our study shed some novel light on miR-128b-PDK1/Akt/NF-κB axis on GC progression.