Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

In the context of diabetes, obesity, and metabolic syndrome, the inflammatory signaling has critical roles in the pathogenesis of nonalcoholic fatty liver disease (NAFLD), but the underlying mechanisms remain poorly delineated. Herein, early and persistently elevated, proinflammatory cytokine HMGB1 expression was detected in a high-fat diet (HFD)-induced NAFLD model in C57BL/6 mice. The expression and extracellular release of HMGB1 was rapidly and dramatically induced by saturated palmitic acid in vitro. HFD-induced inflammatory response and liver function impairment were both mitigated after the inhibition of endogenous HMGB1 by neutralizing antibody in vivo. The up-regulation of HMGB1 was thought to be modified by dual channels: in the transcriptional level, it was regulated by JNK1/JNK2-ATF2 axis; post-transcriptionally, it was regulated by the microRNA (miR)-200 family, especially miR-429. miR-429 liver conditional knockout mice (miR-429Δhep), fed either a normal diet or an HFD, showed severe liver inflammation and dysfunction, accompanied by greater expression of HMGB1. Intriguingly, the up-regulation and release of HMGB1 could in turn self-activate TLR4-JNK1/JNK2-ATF2 signaling, thus forming a positive feedback. Our findings reveal a novel mechanism by which HMGB1 expression was regulated by both the JNK1/2-ATF2 axis and the miR-200 family, which provides a potential new approach for the treatment of NAFLD.-Chen, X., Ling, Y., Wei, Y., Tang, J., Ren, Y., Zhang, B., Jiang, F., Li, H., Wang, R., Wen, W., Lv, G., Wu, M., Chen, L., Li, L., Wang, H. Dual regulation of HMGB1 by combined JNK1/2-ATF2 axis with miR-200 family in nonalcoholic steatohepatitis in mice.

Low-dose Lipopolysaccharide Alleviates Spinal Cord Injury-induced Neuronal Inflammation by Inhibiting microRNA-429-mediated Suppression of PI3K/AKT/Nrf2 Signaling.

This study investigated the impact of low-dose lipopolysaccharide (LPS) on spinal cord injury (SCI) and the potential molecular mechanism. Rats were randomly assigned to four groups: Sham, SCI, SCI + LPS, and SCI + LPS + agomir. Allen's weight-drop method was used to establish an in vivo SCI model. The Basso Bcattie Bresnahan rating scale was employed to monitor locomotor function. An in vitro SCI model was constructed by subjecting PC12 cells to oxygen and glucose deprivation/ reoxygenation (OGD/R). Enzyme-linked immunosorbent assay (ELISA) was applied for the determination interleukin (IL)-1ß and IL-6. The dual luciferase reporter assay was used to validate the targeting of microRNA (miR)-429 with PI3K. Immunohistochemical staining was used to assess the expression of PI3K, phosphorylated AKT and Nrf2 proteins. The Nrf2-downstream anti-oxidative stress proteins, OH-1 and NQO1, were detected by western blot assay. MiR-429 expression was detected by fluorescence in situ hybridization and real-time quantitative reverse transcription PCR. In vitro, low-dose LPS decreased miR-429 expression, activated PI3K/AKT/Nrf2, inhibited oxidative stress and inflammation, and attenuated SCI. MiR-429 was found to target and negatively regulate PI3K. Inhibition of miR-429 suppressed low-dose LPS-mediated oxidative stress and inflammation via activation of the PI3K/AKT/Nrf2 pathway. In vivo, miR-429 was detectable in neurons. Inhibition of miR-429 blocked low-dose LPS-mediated oxidative stress and inflammation via activation of the PI3K/AKT/Nrf2 pathway. Overall, low-dose LPS was found to alleviate SCI-induced neuronal oxidative stress and inflammatory response by down-regulating miR-429 to activate the PI3K/AKT/Nrf2 pathway.

The microRNA-429/DUSP4 axis regulates the sensitivity of colorectal cancer cells to nintedanib.

Colorectal cancer (CRC) is recognized as one of the most common malignancies, which ranks third among all cancer-related deaths worldwide. Nintedanib is an orally available tyrosine kinase inhibitor that can treat CRC; however, drug resistance to nintedanib leads to unsatisfactory treatments for patients with CRC. The aim of the present study was to explore whether overexpression of miR-429 elevated the sensitivity of CRC cells to nintedanib by downregulating dual specificity protein phosphatase 4 (DUSP4). The nintedanib-resistant CRC cell model was established via the treatment of cells with nintedanib in a dose-dependent manner. Reverse transcription-quantitative PCR was used to detect the expression levels of miR-429 and DUSP4, and to confirm the transfection efficiency of miR-429 mimic and DUSP4 overexpression plasmid. Cell Counting Kit-8 assay was utilized to measure the inhibition rate of cells. Western blotting was conducted to observe the expression levels of DUSP4 protein, apoptosis-related proteins and proteins related to the JNK signaling pathway. Dual-luciferase reporter assay was performed to evaluate luciferase activity and TUNEL assay was conducted to detect the apoptosis of cells. The results revealed that miR-429 mimic elevated the sensitivity of CRC cells to nintedanib. Moreover, by ENCORI prediction, DUSP4 was identified as a target gene of miR-429, and overexpression of DUSP4 reversed the inducing effect of miR-429 overexpression on the sensitivity of CRC cells to nintedanib. In conclusion, overexpression of miR-429 may elevate the sensitivity of CRC cells to nintedanib through inhibition of the JNK signaling pathway by targeting DUSP4.These findings may aid in the prevention of drug resistance of CRC cells to nintedanib.

HFD-induced TRAF6 upregulation promotes liver cholesterol accumulation and fatty liver development via EZH2-mediated miR-429/PPARα axis.

Despite the increasing prevalence of fatty liver diseases worldwide, the molecular mechanism underlying their pathogenesis remains poorly defined. This study examines the expression and significance of tumor necrosis factor (TNF) receptor-associated factor 6 (TRAF6) in the high-fat diet (HFD)-induced mouse obesity model and the oleic acid/palmitic acid (OA/PA)-induced cell model. After developing these models, we measured the expressions of TRAF6, enhancer of the zeste homolog 2 (EZH2), and peroxisome proliferator activated receptor alpha (PPARα). The expression of TRAF6, EZH2, and PPARα was manipulated to investigate their roles in cholesterol accumulation through evaluating the plasma levels of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C). Co-immunoprecipitation (coIP) assay was used to determine the interaction between TRAF6 and EZH2 and chromatin immunoprecipitation (ChIP) assay to detect the enrichment of EZH2 and H3K27me3 in microRNA-429 (miR-429) promoter. We found that HFD resulted in elevated TRAF6 and miR-429 in fatty liver and reduced EZH2 and PPARα. TRAF6 mediated the ubiquitination of EZH2 and increased miR-429 expression, and miR-429 targeted PPARα. TRAF6 increased cholesterol accumulation in liver cells in vitro via the EZH2/miR-429/PPARα axis. Collectively, HFD upregulates TRAF6 and ubiquitinates EZH2 to promote the miR-429-dependent inhibition of PPARα, leading to cholesterol accumulation in liver and the occurrence of fatty liver.

MicroRNA-429 acts as a tumor suppressor in colorectal cancer by targeting high mobility group box 3.

Colorectal cancer (CRC) is one of the most common solid tumors worldwide and has an extremely poor prognosis. MicroRNA-429 (miR-429) has been reported to participate in the progression of CRC. However, the pathological mechanisms require further investigation. The aim of the present study was to investigate the association between miR-429 and high mobility group box 3 (HMGB3) in CRC and the associated mechanism. The mRNA expression levels of miR-429 and HMGB3 in 65 paired CRC and adjacent tissues were examined by reverse transcription-quantitative PCR. Furthermore, a dual-luciferase reporter assay was performed to identify the association between miR-429 and HMGB3. Finally, the effects of miR-429 and HMGB3 on the proliferation and apoptosis of CRC cells were detected. As a result, it was identified that miR-429 expression was downregulated and HMGB3 expression was upregulated in CRC tissues compared with in adjacent non-cancer tissues, and the expression levels of miR-429 were negatively associated with those of HMGB3. Notably, HMGB3 was demonstrated to be a direct target of miR-429 by dual-luciferase reporter assay. Furthermore, transfection with a miR-429 mimic significantly inhibited HMGB3 expression and led to decreased proliferation and increased apoptosis of CRC cells. On the other hand, transient overexpression of HMGB3 partially inhibited the antitumor effects of miR-429. To the best of our knowledge, the present study demonstrated for the first time that miR-429 regulated the proliferation and apoptosis of CRC cells via HMGB3, suggesting a specific tumor suppressive function of the miR-429/HMGB3 signaling pathway in CRC.

Berberine inhibits the proliferation, invasion and migration of endometrial stromal cells by downregulating miR­429.

Endometriosis (EM) is a common gynecological disease, and its pathological process is accompanied by the migration and proliferation of uterine cells. Berberine (BBR) has been shown to exhibit antitumor activity; however, the effects of BBR on EM have seldom been reported to date. The expression of microRNA (miR)­429 is upregulated in EM and miR­429 can be used as a target for drug regulation of cancer cells. Whether BBR plays a regulatory role in EM by targeting miR­429 has not been reported. Thus, the aim of the present study was to determine the effects of BBR on EM cells. The survival rate of immortalized human endometrial stromal cells (HESCs) was determined using a Cell Counting Kit­8 assay. A colony formation assay was used to detect the rate of cell proliferation. The expression levels of proliferation­related proteins, including proliferation marker protein Ki­67 (Ki­67) and proliferating cell nuclear antigen (PCNA), were detected by reverse transcription­quantitative PCR (RT­qPCR) and western blotting. Wound healing and Transwell assays were performed to detect cell migration and invasion, and western blotting was used to detect the expression of the migration­ and invasion­related proteins, including matrix metalloproteinase (MMP)2, MMP4 and MMP9. The expression of miR­429 was detected by RT­qPCR following its overexpression via cell transfection. The results revealed that treatment with 80 µM BBR significantly inhibited cell proliferation and colony formation, and inhibited the expression of Ki­67 and PCNA proteins in HESCs. BBR inhibited cell invasion and migration, as well as the expression of MMP2, MMP4 and MMP9. In this process, it was found that the expression of miR­429 decreased following treatment of the cells with BBR, whereas the inhibitory effects of BBR on cell proliferation, invasion and migration were suppressed following the overexpression of miR­429. Overall, the findings of the present study indicated that BBR inhibited the proliferation, invasion and migration of HESCs by downregulating the expression of miR­429.

Epithelial to Mesenchymal Transition(EMT) Regulated by MicroRNA-429 in Pancreatic Ductal Adenocarcinoma (PDAC).

In this study, we aimed to investigate the role of miRNA-429 in the pathogenesis of pancreatic ductal adenocarcinoma(PDAC) and the potential mechanism of this procedure. Totally 95 consecutive patients with PDAC diagnosed by pathology were retrospectively collected from June 1st , 2015 to August 30th, 2019 in Department of General Surgery, Jingmen First People's Hospital. The human pancreatic cancer cell line Bxpc-3 and Panc-1 were used and the cell proliferation and migration were detected by MTT assays and Transwell assays, respectively. The quantitative real-time polymerase chain reaction (qRT-PCR) was applied to evaluate the expression in RNA level. Our results showed that overexpression of miRNA-429 could suppress the cell invasion, proliferation and metastasis through regulating the process of EMT in PDAC cell line, while low expression of miRNA-429 had the opposite effects. We demonstrated that miRNA-429 had critical roles in the pathogenesis of PDAC. Clinically, we observed that tumor tissues from patients with PDAC exhibited significantly decreasing in miRNA-429 expression compared with the non-tumor tissues. Additionally, decreased expression of miRNA-429 in tumor tissues of patients with PDAC was associated with poorer prognosis and several clinical-pathological characteristics. In conclusion, miRNA-429 exerted anti-tumor functions in PDAC through the regulation of EMT process. The results of this study would provide a novel insight into tumorigenesis and the basis for the development of miRNA-targeting therapies against PDAC.

Knockdown of circ­PVT1 inhibits the progression of lung adenocarcinoma and enhances the sensitivity to cisplatin via the miR­429/FOXK1 signaling axis.

Lung cancer is one of the most prevalent cancers in China, and its incidence and morbidity remain high due to various independent factors. Lung adenocarcinoma (ADC) is the most common type of non­small cell lung carcinoma. Circular RNA plasmacytoma variant translocation 1 (circ­PVT1) plays an oncogenic role in various types of cancer, but the specific role of circ­PVT1 in lung ADC has not yet been reported. In the present study, circ­PVT1 was knocked down in A549 cells and the cell viability, proliferation, migration and invasion were measured via MTT, colony formation, wound healing and Transwell assays, respectively. Then, the cell viability of A549 cells with circ­PVT1­knockdown or ­overexpression was detected after exposure to cisplatin (DDP). After confirming the associations among circ­PVT1, microRNA (miR)­429 and forkhead box k1 (FOXK1) using various tools and assays, the cellular functions of A549 cells treated with combined short hairpin (sh)RNA­circ­PVT1 and miR­429 inhibitor/pcDNA3.1­FOXK1 were tested again. The expression of circ­PVT1 was found to be increased in lung ADC cells, and shRNA­circ­PVT1 led to decreased cell viability, proliferation, migration and invasion. The expression of circ­PVT1 was higher in A549/DDP cells than that in A549 cells, and the activity of caspase­3 was also activated by DDP in A549/DDP cells transfected with shRNA­circ­PVT1, whereas it was inactivated by DDP in A549 cells transfected with circ­PVT1 overexpression plasmid. Furthermore, the decreased cell viability, proliferation, invasion and migration induced by shRNA­circ­PVT1 could be abated by transfection with miR­429 inhibitor and pcDNA3.1­FOXK1. In conclusion, interference of circ­PVT1 inhibits the progression of lung ADC and enhances its sensitivity to DDP via miR­429/FOXK1, which may provide a theoretical basis for the use of novel targets in the treatment of lung ADC.

Anti­inflammatory role of microRNA­429 in human gingival epithelial cells­inhibition of IL­8 production through direct binding to IKKß mRNA.

MicroRNAs (miRNAs), a family of small non­coding RNAs, serve a pivotal role in the regulation of the inflammation by modulating the expression of various genes. However, the molecular mechanism by which miRNAs regulate inflammation­associated molecules in oral epithelial cells remains to be elucidated. The present study examined the biological function of miR­429 by performing the gain­/loss­of­function studies of miR­429 in a gingival squamous cell carcinoma line Ca9­22 cells that either over­ or under­expressed miR­429 through transient transfection with miR­429 mimic or miR­429 inhibitor, respectively. The results demonstrated that the over­expression of miR­429 suppressed the mRNA level of several interleukins, including IL­8. In addition, the over­expression of miR­429 reduced IL­8 secretion under the basal and TNF­α stimulated conditions, whereas the secretion of IL­8 was enhanced when miR­429 was under­expressed. The over­expression of miR­429 inhibited the activation of the transcription factor NF­κB. Furthermore, we found that miR­429 suppressed both mRNA and protein levels of IKKß via its direct binding to the 3'­untranslated region of IKKß mRNA. In addition, the downregulation of IKKß by small interfering RNA reduced both NF­kB activity and IL­8 production in Ca9­22 cells. Taken together, the findings revealed the molecular mechanism of miR­429 to regulate the inflammatory mediator in gingival cells and suggested that it could be useful as a therapeutic target of oral inflammatory diseases.

hsa­miR­429 targets CBX8 to promote cell apoptosis in diffuse large B­cell lymphoma.

Diffuse large B­cell lymphoma (DLBCL) is the most common type of non­Hodgkin lymphoma worldwide. Several studies have indicated that Homo sapiens (hsa)­microRNA (miR)­429 exerts a tumor­suppressive effect on a variety of malignant tumors. To the best of our knowledge, the molecular function and mechanism of action of hsa­miR­429 in DLBCL have not been evaluated to date. The present study demonstrated that the expression of hsa­miR­429 in DLBCL cells was significantly reduced. hsa­miR­429 inhibited the proliferation of the DLBCL cell lines, SUDHL­4 and DB, and promoted apoptosis. A dual luciferase reporter assay was used to demonstrate that chromobox 8 (CBX8) was the target gene of hsa­miR­429. Overexpression of CBX8 promoted the proliferation of SUDHL­4 and DB cells and inhibited apoptosis, thereby playing a cancer­promoting role. Transfection of hsa­miR­429 mimic into DB cells overexpressing CBX8 antagonized the effect of CBX8 on the proliferation of DB cells. Moreover, the apoptotic rate was increased in DB cells overexpressing CBX8 and transfected with hsa­miR­429 mimic, while the proportion of cells in the G2/M phase was significantly reduced. These results demonstrated the antagonistic effect of hsa­miR­429 on the oncogenic function of CBX8. Therefore, in DLBCL, the tumor suppressor effect of hsa­miR­429 may be achieved by targeted downregulation of CBX8, suggesting that hsa­miR­429 may be used as a diagnostic marker and a potential nucleic acid drug for DLBCL. CBX8 may also represent an effective therapeutic target for DLBCL.

MicroRNA-429 inhibits cancer cell proliferation and migration by targeting AKT1 in renal cell carcinoma.

MicroRNAs (miRNAs or miR) serve as oncogenes and tumor suppressors. In a previous study, it was revealed that has-miRNA-429 (miR-429) is a tumor suppressor in 786-O renal cell carcinoma (RCC) cells. However, its mechanism in RCC remains to be determined. The present study aimed to explain the functional role and mechanism of miR-429 in RCC pathogenesis. Luciferase reporter assays demonstrated that miR-429 overexpression reduced the transcriptional activity of AKT serine/threonine kinase 1 (AKT1). Reverse transcripton-quantitative (RT-q) PCR and western blot analysis indicated that the mRNA and protein expression of AKT1 was downregulated in 786-O RCC cell lines when miR-429 was overexpressed, indicating that miR-429 may directly target AKT1 in RCC. Therefore, miR-429 overexpression enhanced the inhibition of tumor size and weight in nude mice in vivo. The current study indicated that the novel miR-429-regulated pathway may provide insights into RCC oncogenesis and metastasis.

MicroRNA-429 inhibits bone metastasis in breast cancer by regulating CrkL and MMP-9.

Bone metastasis is common in late-stage breast cancer patients and leads to skeletal-related events that affect the quality of life and decrease survival. Numerous miRNAs have been confirmed to be involved in metastatic breast cancer, such as the miR200 family. Our previous study identified microRNA-429 (miR-429) as a regulatory molecule in breast cancer bone metastasis. However, the effects of miR-429 and its regulatory axis in the metastatic breast cancer bone microenvironment have not been thoroughly investigated. We observed a positive correlation between miR-429 expression in clinical tissues and the bone metastasis-free interval and a negative correlation between miR-429 expression and the degree of bone metastasis. We cultured bone metastatic MDA-MB-231 cells and used conditioned medium (CM) to detect the effect of miR-429 on osteoblast and osteoclast cells in vitro. We constructed an orthotopic bone destruction model and a left ventricle implantation model to examine the effect of miR-429 on the metastatic bone environment in vivo. The transfection experiments showed that the expression levels of V-crk sarcoma virus CT10 oncogene homolog-like (CrkL) and MMP-9 were negatively regulated by miR-429. The in vitro coculture experiments showed that miR-429 promoted osteoblast differentiation and that CrkL promoted osteoclast differentiation. The two animal models showed that miR-429 diminished local bone destruction and distant bone metastasis but CrkL enhanced these effects. Furthermore, CrkL and MMP-9 expression decreased simultaneously in response to increased miR-429 expression. These findings further reveal the possible mechanism and effect of the miR-429/CrkL/MMP-9 regulatory axis in the bone microenvironment in breast cancer bone metastasis.

Down-regulation of microRNA-429 alleviates myocardial injury of rats with coronary heart disease.

In recent years, the impact of microRNAs (miRNAs) on coronary heart disease (CHD) has been identified. This study was aimed to investigate the regulative role of microRNA (miR-429) in myocardial injury of rats with CHD. Expression of miR-429 in CHD patients and healthy people was detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR). The CHD rat models were injected with normal saline, mimics negative control (NC), miR-429 mimics, inhibitors NC and miR-429 inhibitors twice a week, for 4 weeks. Levels of inflammatory factors, oxidative stress indices as well as apoptosis of cardiomyocytes were determined by a series of assays. Expression of miR-429 was up-regulated in CHD patients. Reduced miR-429 could decline the expression of oxidative stress-related factors and inflammation-related factors, and inhibit the apoptosis of cardiomyocytes in rats with CHD. Moreover, the down-regulation of miR-429 could promote the expression of CrkL and repress activation of the MEK/ERK signaling pathway. This study reveals that restrained miR-429 could exert a protective impact on myocardial injury of rats with CHD by suppressing oxidative stress, inflammation reaction and apoptosis of cardiomyocytes. The function mechanisms may relate to the up-regulation of CrkL and inhibition of the MEK/ERK signaling pathway.

Long non-coding RNA MALAT1 sponges microRNA-429 to regulate apoptosis of hippocampal neurons in hypoxic-ischemic brain damage by regulating WNT1.

Hypoxic-ischemic brain damage (HIBD) is a common neurological disorder. Emerging reports reveal that long non-coding RNAs and microRNAs (miRs) are implicated in the progress of HIBD. In this study we tried to ascertain whether lncRNA MALAT1, with the involvement of miR-429 and WNT1, affects HIBD. Initially, a HIBD mouse model was established. Then, we treated HIBD mice with dexmedetomidine (DEX) and then up- or down-regulated the expression of MALAT1, miR-429 and WNT1 in HIBD mice and neurons. Meanwhile, brain injury and hippocampal neuronal apoptosis were evaluated. Moreover, the interaction among MALAT1, miR-429 and WNT1 in HIBD was investigated. MALAT1 and WNT1 were high-expressed in brain tissues of HIBD mice while miR-429 was low-expressed in brain tissues from HIBD mice. Interestingly, MALAT1 silencing was observed to enhance the cerebral protection of DEX against HIBD. In addition, it was confirmed that MALAT1 sponged miR-429 downregulating expression of miR-429, thereby promoting apoptosis of hippocampal neurons. This effect was achieved through up-regulating the level of WNT1. Taken together, this study demonstrates that silencing of MALAT1 enhances the cerebral protection of DEX against HIBD by suppressing WNT1 expression through miR-429.

MicroRNA-429 decreases the invasion ability of gastric cancer cell line BGC-823 by downregulating the expression of heparanase.

The present study aimed to measure the expression of microRNA (miRNA/miR) -429 in gastric cancer and investigate the associated mechanism of action. A total of 30 patients with gastric cancer who received radical or palliative resection at Jining No. 1 People's Hospital between January-October 2016 were included in the present study. Resected gastric cancer and tumor-adjacent tissues were resected. Gastric cancer BGC-823 cells were transfected with miR-429 mimics to induce the overexpression of miR-429, or transfected with small interfering (si)RNA of heparanase (HPSE) for the silencing of HPSE. Reverse transcription-quantitative polymerase chain reaction was used to measure the expression of miR-429 or HPSE mRNA. Western blotting was employed to determine the protein expression of HPSE. Cell-Counting Kit-8 assay was carried out to test cell proliferation and a Transwell assay was used to determine cell invasion ability. Expression of HPSE mRNA and protein in gastric cancer tissues was increased compared with tumor-adjacent tissues. Reduced expression of miR-429 in gastric cancer tissues may be associated with the targeting of HPSE mRNA by miR-429. Overexpression of miR-429 inhibited the transcription and translation of the HPSE gene. However, overexpression of miR-429 did not affect the proliferation of gastric cancer cells. Notably, overexpression of miR-429 reduced the invasion ability of gastric cancer cells. Transfection with HPSE siRNA decreased the expression of the HPSE protein in BGC-823 cells and inhibited the occurrence and development of gastric cancer by reducing the invasion ability of the cells. The present study demonstrated that expression of miR-429 in gastric cancer tissues was significantly reduced compared with tumor-adjacent tissues. As a tumor-suppressor gene, miR-429 decreases the invasion ability of gastric cancer cells by downregulating the expression of HSPE.

miR-429 expression in bladder cancer and its correlation with tumor behavior and clinical outcome.

We previously showed that microRNA-429 (miR-429) played an important role in epithelial-mesenchymal transition (EMT) of urothelial cell carcinoma of the bladder. We herein evaluated the expression of miR-429 in bladder cancer and its potential relevance to clinicopathological characteristics and patient survival. Relative expression levels of miR-429 in surgical bladder cancer tissue specimens obtained from 76 patients with bladder cancer were measured by chromogenic in situ hybridization. miR-429 expression was significantly higher in specimens from alive patients than expired patients in both of 5-year overall survival (OS) (0.59 ± 0.09 vs. 0.27 ± 0.12; p < 0.05) and 5-year recurrence-free survival (RFS) (0.63 ± 0.10 vs. 0.33 ± 0.10; p < 0.05). The univariate Cox proportional hazards analysis revealed that tumor grade, stage, and miR-429 expression were significantly associated with patient survival. In multivariate analysis, tumor stage and miR-429 expression were significantly associated with 5-year OS (hazard ratio [HR] 4.70, p < 0.001) and 5-year-RFS (HR 2.20, p < 0.05). The Kaplan-Meier analysis showed that patients with miR-429 expression had significantly better 5-year OS and 5-year RFS rates than those without miR-429 expression (84.4% vs. 61.4%, p < 0.05 and 71.9% vs. 45.5%, p < 0.05, respectively). miR-429 may be considered as an adjunctive prognostic marker in addition to tumor grade and stage in bladder cancer.

MicroRNA-429 inhibits gastric cancer migration and invasion through the downregulation of specificity protein 1.

microRNAs (miRs) have been reported to have an important role in tumorigenesis and tumor progression. Although miR-429 has been shown to be downregulated in gastric cancer (GC), the function of miR-429 in the metastasis of GC has yet to be investigated. In the present study, GC cells were transfected with miR-429, and reverse transcription-quantitative polymerase chain reaction, cell migration assays, cell invasion assays, western blot analysis and luciferase assays were conducted to investigate the role of miR-429 in GC cells. It was demonstrated that miR-429 expression was markedly increased following transfection of the cells with miR-429. Furthermore, miR-429 was shown to inhibit the migration and invasion of GC cell lines. In addition, this study provided evidence that miR-429 directly targets specificity protein 1 in GC cells. The results of the present study may enhance current knowledge regarding the molecular basis of cancer metastasis and provide a potential therapeutic strategy for GC.

Advances in biological functions of miR-429 / 解剖学报

MicroRNAs (miRNAs, miR) are endogenous non-coding single-stranded RNAs with a length of about 22 nucleotides. These RNAs play an important biological function within cells, among which, miR-429 has been proved to play an important role in inhibiting tumor development and tumor progression as well as in cell differentiation and neurological diseases by regulating the expression of different target genes. In this paper, the role of miR-429 and its downstream targeted genes in cell proliferation, apoptosis, migration and invasion is summarized.

The Potential Prognostic Value of MicroRNA-429 for Human Gliomas.

MicroRNA-429 (miR-429) is reported to be frequently dysregulated in cancer. Studies have demonstrated that miR-429 functions as either an oncogene or a tumor suppressor depending on the tumor type. To date, its role in human glioma has not yet been reported. The aim of this study was to investigate the expression levels, clinicopathological significance, and prognostic significance of miR-429 in glioma tissues. Quantitative real-time PCR assay was performed to detect miR-429 expression in human glioma and non-neoplastic brain tissues. The association of miR-429 expression with clinicopathological features and prognoses of glioma patients were analyzed by SPSS 17.0 statistical software. The expression levels of miR-429 were found to be distinctly increased in glioma tissues compared to non-neoplastic brain tissues (P<0.01) with ascending pathological grade. The 5-year survival rate of patients with high miR-429 expression was significantly lower than those with low miR-429 expression (P<0.001). Moreover, multivariate Cox regression analyses showed that miR-429 high expression is an independent prognostic factor for glioma patients. Taken together, miR-429, was significantly up-regulated in glioma tissues. Therefore, miR-429 could be considered as an independent prognostic factor and potential therapeutic target for glioma.

MiR-429 reverses epithelial-mesenchymal transition by restoring E-cadherin expression in bladder cancer.

Epithelial-mesenchymal transition (EMT) accompanying loss of E-cadherin is important for invasiveness and metastasis of bladder cancer. MicroRNAs (miRs) had been associated with cancer progression and differentiation in several cancers. Our goal is to find out the specific miR which modulates EMT in bladder cancer. Real-time quantitative polymerase chain reaction was used to measure the miRs expression in urothelial cell carcinoma (UCC) cell lines. MiR or siRNA mimics was used to regulate miR and mRNA level respectively. Migration and scratch assays were used to determine the migratory ability. Zymography assay was used to confirm the metalloproteinase activity. Western blotting was used to elucidate the mechanism which regulated by specific miR. MiR-429 was highly expressed in low grade UCC cell lines. Exogenous mimic of miR-429 treatment dramatically inhibited the migratory ability of T24 cells. MiR-429 downstream target ZEB1 was decreased, E-cadherin was restored, and ß-catenin was contrarily decreased by exogenous mimic of miR-429 treatment in T24 cells. Cell invasive ability was also inhibited by exogenous mimic of miR-429 treatment through inactivating the MMP-2 activity in T24 cells. E-cadherin protein expression level was inhibited by E-cadherin siRNA accompanied with increasing cell migratory ability when compared with control group in low grade TSGH8301 cells. MiR-429 decreased the cell migratory and invasive abilities through reducing ZEB1 and ß-catenin, restoring the E-cadherin expression and inactivation of MMP-2 of UCC cells. MiR-429 might be used as a progression marker of bladder cancer.