14 publications mentioning hsa-mir-592

Open access articles that are associated with the species Homo sapiens and mention the gene name mir-592. Click the [+] symbols to view sentences that include the gene name, or the word cloud on the right for a summary.

1

Overexpression of WSB1 protein in HCC cells also significantly abolished the suppression of miR-592 on glucose consumption (Figure 4D), lactate production (Figure 4E), and ATP level (Figure 4F), suggesting that miR-592 may suppress glycolysis by directly decreasing WSB1 expression in HCC cells.

It was found that the overall survival of HCC patients with low miR-592 expression was shorter than that of patients with high miR-592 expression (Figure 1E) Together, our data indicate that downregulation of miR-592 expression closely associates with poor HCC prognosis, suggesting that miR-592 participates in HCC progression.

Therefore, although miR-592 overexpression or WSB knockdown increased pVHL stability, CoCl [2] inhibited PHD activity and disrupted the interaction between PHD and pVHL, thus miR-592 overexpression or WSB knockdown could not block the CoCls -induced lactate production.

Herein, we report that the expression of miR-592 in HCC cells and tissues is downregulated, and miR-592 directly targets WSB1 mRNA.

It was found that overexpression of miR-592 remarkably suppressed the luciferase activity of the reporter gene with the wild-type construct, but not with the mutant WSB1 3′-UTR construct in both SK-Hep-1 and SMMC7721 cells (Figure 3B), suggesting that miR-592 directly targeted the WSB1 3′-UTR.

Although it was observed thatthe decreasing of lactate production upon overexpression of miR-592 or knockdown of WSB1 statistically significant compared to control in DMSO treated group, neither miR-592 overexpression nor WSB1 knockdown blocked the CoCl [2] or HIF-1α -induced increase of HK2 and Glut1 expression and lactate production (Figure 5F, lower panel).

In addition, Given that CoCl [2] only partially rescued miR-592 overexpression- caused cell proliferation retardation, and so did WSB-1 overexpression, this suggests that there may be alternative targets of miR592 that contributed to the growth retardation effect of miR-592.

Knockdown of miR-592 also increased the expression of Glut1 and HK2 and lactate production; however, YC-1 rescued Glut1 and HK2 expression and lactate production enhanced by miR-592 knockdown (Figure 5E).

Our results indicate that miR-592 is a robust inhibitor of the Warburg effect by targeting WSB1/HIF-1a axis, and more importantly, the newly identified miR-592 and its downstream pathway might serve as potential therapeutic targets and novel prognosis indicators in HCC.

Indeed, we observed that the miR-592 knockdown -mediated upregulation of the WSB1 level increased the expression of HIF-1α, which plays an important role in critical aspects of HCC cancer biology, especially glucose metabolism [16, 17].

In the present study, we showed that miR-592, by suppressing WSB1, but not key glycolytic genes, Myc, HK1 HIF-1α, HK2, stat3, or PKM2, effectively inhibits both glucose metabolism in HCC cells and the growth of xenografted HCC tumors, indicating a strong rationale for developing miR-592 as a novel metabolism -targeting therapeutic agent against HCC.

These data further support mechanism postulating that miR-592 knockdown directly promotes the expression of WSB1, thereby enhancing HIF-1α protein expression to maintain increased glycolysis in HCC (Figure 6E).

Together, our findings indicate that HIF-1α is responsible for the changes in glycolysis induced by miR-592 downregulation or WSB1 upregulation.

According to the median level of miR-592 expression, the patients (90 cases) were divided into two groups as either high expression (45 patients) or low expression (45 patients).

In addition, miR-592 knockdown increased, whereas miR-592 overexpression decreased HIF-1α protein expression, but not mRNA level (Figure 5D).

F. The effect of CoCl2 on the expression of HK2 and Glut1 (left panel), and the effects of CoCl2 on lactate production (right panel) in SK-Hep-1 cells with WSB1 knockdown or miR-592 overexpression.

Moreover, the inhibitory role of miR-592 in proliferation and cell cycle progression was reverted under the condition of mutant WSB1 overexpression (Figure 3E and Figure 3F).

Conversely, suppression of miR-592 enhanced the expression of WSB1 in both SK-Hep-1 and SMMC7721 cells (Figure 3C).

To uncover the underlying mechanism accounting for miR-592 -induced inhibition of HCC cells proliferation, bioinformatics research was performed to predict miR-592 targets using PITA and miRanda [11].

In order to validate the therapeutic value of miR-592, tumor cells with miR-592 overexpression were injected into mice to examine the inhibition of tumor growth.

In addition, two recent studies indicating that miR-592 inhibits prostate cancer and colorectal cancer growth [27, 28], further support miR-592 as a strongly general target candidate for anticancer therapies.

Our data suggest that decreased WSB1/HIF-1α expression via overexpression of miR-592 may decrease proliferation, glycolysis, and lactate production in HCC cells, which provides a new approach for HCC therapeutics.

As shown in supplementary Figure 1A, 1B, 1C, 1D and 1E, miR-592 overexpression significantly inhibited HCC growth in mice mo dels.

B. Correlationship between miR-592 expression and WSB1 and HIF-1a expression in HCC patients tissues.

To investigate whether the tumor suppressor role of miR-592 on HCC cell proliferation is mediated by inhibiting the expression of WSB1, SK-Hep-1 cells were co -transfected with miR-592 mimics and WSB1 plasmid without 3′-UTR.

As shown in Figure 6A, levels of both WSB1 and HIF-1α in human HCC tissues with high miR-592 expression were lower than those in HCC with low miR-592 expression.

In conclusion, the present study demonstrated a novel inhibitory role of miR-592 in HCC glycolysis by targeting WSB1/HIF-1α axis.

Consistently, we observed an inverse expression pattern between miR-592 and WSB1 or HIF-1α, whereas a positive expression pattern between WSB1 and HIF-1a was observed (Figure 6B).

To uncover the molecular mechanism of miR-592 HCC inhibitory potential, we hypothesized that decreased WSB1 expression by miR-592 would reduce the HIF-1α level in HCC because HIF-1α signaling has been demonstrated to be enhanced by WSB1 in various cancers [10].

E. The expression of HK2 and Glut1 in SK-Hep-1 cells treated with YC-1 and/or miR-592 knockdown (left panel); lactate production in HCC (right panel).

As shown in Figure 3D, miR-592 -induced WSB1 downregulation was rescued following co-transfection.

D. The protein level (left panel) or mRNA level (right panel) of HIF-1α with overexpression or knockdown of miR-592.

miR-592 is frequently downregulated in human HCC tissues and cell lines, which associates with low postoperative survival rate.

B. Measurement of the cellular G6P level in HCC cells with miR-592 overexpression or downregulation.

The new finding of this study is that downregulation of miR-592 may facilitate HCC pathogenesis through the WSB1/HIF-1α axis.

Consistently, overexpression of miR-592 greatly reduced the cellular G6P level in HCC cells, whereas knockdown of the endogenous miR-592 increased the G6P level (Figure 4B).

miR-592 directly targets WSB1 in HCC cells.

Another important aspect of the current study is the novel role of miR-592 in inhibition of glycolytic phenotypes.

Given that WSB1 increases one of the most important glycolytic proteins-HIF-1α and contributes to human malignancy [10], thus WSB1 was predicted as a potentially functional target gene of miR-592.

As shown in Figure 6C and Figure 6D, the inhibitory role of miR-592 in proliferation and cell cycle progression was reverted under the condition of treatment with CoCl [2].

WSB1 is a bona fide target gene of miR-592 and functional downstream of miR-592 in HCC cells.

Compared with the negative controls, overexpression of miR-592 in SK-Hep-1 and SMMC7721 cells significantly decreased cell number (Figure 2C and Figure 2D) whereas knockdown of the endogenous miR-592 markedly increased the viability of these cells (Figure 2E and Figure 2F).

A. Representative images of IHC staining for WSB1 and HIF-1a in HCC patients with high and low miR-592 expression.

In addition, as shown in supplementary Figure 1F, miR-592 inhibits the migration of HCC cells in vitro.

Therefore, the newly identified miR-592/WSB1/HIF-1α axis provides new insight into the pathogenesis of HCC, suggesting a novel, potential therapeutic target for the treatment of HCC.

E. miR-592 expression was inversely associated with the overall survival of 90 HCC patients.

K. The expression of miR-592 in tumor tissues was analyzed by qRT-PCR.

The miR-592/WSB1 signal pathway inhibits glycolysis in HCC cells.

Figure 6 A. Representative images of IHC staining for WSB1 and HIF-1a in HCC patients with high and low miR-592 expression.

Figure 1 A. qRT-PCR analysis of miR-592 expression in 40 pairs HCC tissues and their adjacent normal tissues.

Clinical significance of miR-592, WSB1 and HIF-1a expression in patients with HCC.

To determine the expression of miR-592 in HCC, we analyzed 40 cases of HCC tissues and adjacent non-tumorous liver tissues using quantitative real-time PCR.

Given that WSB1 promotes VHL ubiquitination and proteasomal degradation, thereby stabilizing HIF under both normoxic and hypoxic conditions, and HIF-1α has been shown to be a key transcription factor promoting glycolytic energy metabolism in cancer cells [10], we wondered if this newly discovered miR-592/WSB1 axis might inhibit glycolysis in HCC cells.

Moreover, overexpression of miR-592 also repressed the protein levels of WSB1 in both SK-Hep-1 and SMMC7721 cells.

The miR-592/WSB1 axis inhibits glycolysis in HCC cells.

D. SK-Hep-1 cells were co -transfected with miR-592 mimics or NC and WSB1 plasmid without 3′-UTR, the expression levels of WSB1 were analyzed using western blot.

Because miR-529 presents as the sole upstream regulator of WSB1, it will be possible that mir-592 is a potential key regulator keeping the equilibrium of WSB1/HIF-1α signaling.

miR-592 was recently proposed to be a new prognosis predictor for HCC patients and a new prospective target for HCC [14, 15].

Clinical relevance of miR-592, WSB1 and HIF-1a expression in patients with HCC.

MiR-592 is frequently down-regulated in human HCC and associates with poor clinicopathologic features.

To demonstrate whether the 3′-UTR of WSB1 could be a genuine target of miR-592, a dual-luciferase reporter system was constructed.

The miR-592 inhibitors (Cat.

The expression of miR-592 was confirmed by qRT-PCR.

A. qRT-PCR analysis of miR-592 expression in 40 pairs HCC tissues and their adjacent normal tissues.

The significance of this study is also underscored by the fact that miR-592 is frequently decreased in human HCCs, and that miR-592 knockout significantly promoted mouse HCC development.

qRT-PCR analysis of tumor tissues confirmed that the expression of miR-592 was significantly decreased (Figure 2K).

As shown in supplementary Figure 2A and 2B, miR-592 significantly decreases HIF-1α stability which was reverted by WSB1 overexpression.

C. The expression of miR-592 in HCC tissues, tissues with lymph node metastases, and their adjacent normal tissues, n=10/group.

In addition, we analyzed the correlationship between miR-592 expression and the patients' prognoses.

B. The expression of miR-592 in HCC tissues was significantly lower than in corresponding normal tissues.

Does HIF-1α as a transcription factor increase or decrease the expression of miR-592 in HCC?

Knockdown of miR-592 enhances glycolysis by increasing WSB1 -induced HIF-1α stability.

Because WSB1 can increase the stability of HIF-1α protein [10] and HIF-1α is one of key transcription factors in enhancing glycolysis [12], we hypothesized that HIF-1α may dictate the miR-592 knockdown -induced glycolysis in HCC cells.

To confirm this role of miR-592 applied in vivo, we tested whether miR-592 knockdown would promote tumour growth in SK-Hep-1 orthotopic tumour mo dels and mouse subcutaneous xenograft mo dels.

E. Schematic diagram of the regulatory pathway from miR-592/WSB1 / HIF-1a axis to glycolysis in HCC.

MiR-592 inhibits HCC cell proliferation and tumor growth.

Moreover, compared with primary HCC tissues and the adjacent normal tissues, tissues from lymph node metastases expressed lower levels of miR-592 (Figure 1C).

Consistent with these clinical findings, the expression of miR-592 was significantly decreased in five HCC cell lines compared with two normal liver cell lines (LO2 and 7701) (Figure 1D).

Compared with the control tissues, the expression of miR-592 was significantly decreased in 33 of 40 HCC tissues (Figure 1A).

To further confirm that miR-592 decreases HIF-1α stability via WSB1, the protein degradation of endogenous HIF-1α was measured in HCC cells with overexpression and knockdown of miR-592 and WSB1.

The effect of miR-592 and CoCl2 on C. SK-Hep-1 cell proliferation and D. cell cycle progression.

Fold changes for the expression levels of miR-592 were calculated using the comparative cycle threshold method (2 [−ΔCT]).

HSTUD0817, Sigma), miR-592 mimics (Cat.

C. and D. Overexpression of miR-592 significantly decreased cell proliferation of SK-Hep-1 and SMMC772 cells compared to the negative controls using CCK-8 assay.

Importantly, reintroduction of WSB1 reverted the miR-592 -induced decrease in the cellular G6P level in HCC cells (Figure 4C).

To further validate the inverse relationship between WSB1, HIF-1α and miR-592 in HCC cells, we examined miR-592, WSB1 and HIF-1α levels in the same human HCC tissues.

SK-Hep-1 cells transfected with anti-miR-592 or negative control (anti-NC) were subcutaneously injected into the flank region of nude mice.

Figure 3 A. Sequence alignment of wild-type (WT) and mutated (Mut) putative miR-592 -binding sites in the 3′-UTR of WSB1.

Given that many of the HCC patients are often being excluded from chemotherapy or surgical resection due to their poor liver functions, thus, therapeutic against miR-592 may selectively provide a novel approach.

A. Sequence alignment of wild-type (WT) and mutated (Mut) putative miR-592 -binding sites in the 3′-UTR of WSB1.

WSB1 3′-UTR sequences containing wild type or mutated binding site of miR-592 were cloned into the psiCHECK2 vector, respectively, and co -transfected with the miR-592 mimics or NC into HCC cells.

Collectively, our findings suggest that miR-592 decreases HCC cell proliferation in vitro and tumorigenicity in vivo.

L. and M. Cell cycle distribution of SK-Hep-1 and SMMC772 cells transfected with miR-592 mimics or NC was examined using flow cytometry.

As shown in Figure 2L and Figure 2M, compared to cells transfected with negative controls, there was a significant increase in G0/G1 phase cells and decrease in S phase cells in miR-592 mimics -transfected SK-Hep-1 and SMMC772 cells, whereas miR-592 knockdown cells displayed the opposite data (data not shown).

And the median level of miR-592 in HCC tissues was obviously lower than in corresponding normal tissues (Figure 1B).

A. and B. SK-Hep-1 and SMMC772 cells were transfected with hsa-miR-592 mimics, anti-miR-592 or negative control, respectively.

To further investigate whether the tumor suppressor role of miR-592 on HCC cells proliferation is mediated through disrupting the enhanced stability of HIF-1α, SK-Hep-1 cells were co -transfected with miR-592 mimics and treated with or without CoCl [2].

However, the molecular mechanisms by which miR-592 exerts its role in tumor are not frequently studied.

Together, these results suggest that WSB1/HIF-1α axis is functional downstream of miR-592 in HCC.

B. Relative luciferase activities of plasmids carrying WT or mut WSB1 3′-UTR in SK-Hep-1 and SMMC7721 cells cotransfected with miR-592 mimics or NC.

In this study, for the first time, we investigated the aberrant status of miR-592 and its potential target gene to explore this miRNA's cancer -associated functions in HCC.

Secondly, our in vivo orthotopic tumour study also further reinforced our in vitro results, and established the potential significance of miR-592 in HCC progression (Figure 2J).

To measure the expression levels of mature miR-592, real-time quantitative RT-PCR was performed.

Stably transfected cells (1.5 × 10 [6] in 0.2 mL) transfected with anti-miR-592 or negative control were injected subcutaneously into the flank region of 6 week-old male severe combined immunodeficiency mice (SCID; Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences).

Figure 2 A. and B. SK-Hep-1 and SMMC772 cells were transfected with hsa-miR-592 mimics, anti-miR-592 or negative control, respectively.

In future studies, it will be interesting to determine whether this newly discovered miR-592/WSB1/HIF-1α axis is distinct in HCC etiology.

As shown in Figure 3A, one predicted binding site in the WSB1 3′-UTR with a perfect complementarity to the seed region of the miR-592 was identified.

The reaction mixture was used for real-time RT-PCR of miR-592.

The effect of miR-592 and WSB1 on E. HCC cell proliferation and F. cell cycle progression.

2

When analyzing the expression levels of the microRNAs selected for validation (Table 2), we observed that 5 of them (hsa-miR-135b *, hsa-miR-592, hsa-miR-31, hsa-mir-135b, hsa-miR-944) have similar patterns of expression, i. e., there are statistically significant differences between the UAs and the control group.

On the SA group, we observed differences in the expression between SAs and the control group in seven out of the 13 microRNAs selected for biological validation (p <0.05), (hsa-miR-135b *, hsa-miR- 489, hsa-miR-592, hsa-miR-369-5p, hsa-miR-31, hsa-mir-135b, hsa-miR-944).

Liu et al. [39] described this overexpression of miR-592 in colorectal cancer associating it to the size of the tumor, the distance of the metastases and the patient survival.

With these data, we consider that alterations in miR-592 expression in ameloblastomas would be mainly linked with tumor growth and progression.

When analyzing the expression of each miRNA, we observed statistically significant differences in 6 microRNAs among the UAs and the control group, (p <0.05) (hsa-miR-135b *, hsa-miR-592, hsa-miR-31, hsa-mir-135b, hsa-miR-944, hsa-miR-142-5p)).

MicroRNAs with a differential expression in both SA and UA when compared to the control group are the microRNA miR-135b, miR-135b *, miR-31, miR-592 and miR-944.

Different studies [36, 37] linked the alterations of miR-592 to neoplastic development in humans.

After applying the inclusion criteria (|FC| <0.2 or> 5 and p adjusted <0.05), as previously mentioned, biological validation was performed by RT-qPCR of the 13 differently regulated miRNAs (hsa-miR-9, hsa-miR-135b*, hsa-miR-194*, hsa-miR-489, hsa-miR-592, hsa-miR-369-5p, hsa-miR-876-5p, hsa-miR-31, hsa-miR-135b, hsa-miR-211, hsa-miR-944, hsa-miR-142-5p, hsa-miR-455-3p), in an independent set of 46 samples corresponding to 19 SA, 8 UA and 19 controls.

miR-592 and miR-552 can distinguish between primary lung adenocarcinoma and colorectal cancer metastases in the lung.

3

NRF2 mediated drug resistance has been shown in colon cancer and is up-regulated in hepatocyte carcinoma [38, 52], indicating a possible link between the downregulation of miR-592 in these disorders and NRF2 activity.

Interestingly, miR-592 down-regulation has also been observed in colon cancers with deficient DNA mismatch repair (dMMR), a common mutation in colorectal tumors.

miR-592 was also down-regulated in hepatitis B hepatocellular carcinoma [89].

For example, in colorectal cancer, low levels of miR-592 expression have shown promise as a biomarker, identifying patients who will react poorly to anti-EGFR chemotherapies [88].

miRNA Location Targets References miR 193b/365Chr16,14397824-14397906 (miR193b)14403142-14403228 (miR-365)TTf1 - oncogenicBCL2 – TSGCyclin D- TSG, uPa(Qi et al., 2012)(Nie et al., 2012)(Li et al., 2009) Chr7, 130562218-130562298Sp-1MCL-1 - oncogenicTCL1 – oncogenic(Amodio et al., 2012)(Mott et al., 2007)(Pekarsky et al., 2006) Chr19, 13985513-13985622SIRT1- oncogenic and TSGKRAS - oncogenicTGFβ - TSGTNF - TSGNOTCH - oncogenic and TSG(Zhang et al., 2012)(Schonrock et al., 2012)(Hashimoto et al., 2010) miR-617 Chr12, 81226312-81226408 N/A miR-592 Chr7, 126698142-126698238 N/A miR-1207 Chr8 129061398-129061484.

miR-592 is located on chromosome 7 at position 126698142-126698238 and has been associated with a number of malignancies including, colorectal and liver carcinoma.

4

Moreover, miR-138 reportedly induced cell cycle arrest by targeting CCND3 in hepatocellular carcinoma cells [37] and miR-592 inhibited cell proliferation by suppressing CCND3 expression in CRC [38] Similarly, miR-4779 efficiently induced cell cycle arrest and apoptosis by targeting CCND3, and further contributed to cell cycle arrest by suppressing PAK2 expression.

Liu Z MiR-592 inhibited cell proliferation of human colorectal cancer cells by suppressing of CCND3 expressionInt.

5

In addition, miR-552 and miR-592 expression was up-regulated in pMMR tumors and down-regulated in dMMR tumors.

Significant differences were also seen in 6 miRNAs including miR-31 and miR-592, in the direct comparison of tumors that were deficient or proficient for mismatch repair.

miRNAs that showed decreased levels in pMMR relative to dMMR tumors included miR-552, miR-592, miR-181c and miR-196b.

Figures Array data was validated by by qRT-PCR for 10 miRNAs (mir-1, miR-10b, miR-135b, miR-147, miR-31, miR-33, miR-503, miR-552, miR-592, miR-622).

6

One might hypothesize that the SNP may potentially affect posttranslational modifications of GRM8 via regulating the expression of microRNA592.

7

miR-592 functioned as a tumor suppressor in lung cancer by targeting SOX9 [29].

8

They characterized low expression of hsa-miR-592 and high expression of hsa-miR-10b-5p and hsa-miR-615-3p were associated with tumors located in the right colon relative to the left colon and rectum and high expression of hsa-miR-615-3p [14].

9

Twelve miRNAs (hsa-miR-130b, hsa-miR-203, hsa-miR-1974, hsa-miR-592, hsa-miR-200a, hsa-miR-429, hsa-miR-183, hsa-miR-182, hsa-miR-1290, hsa-miR-141, hsa-miR-135b, and hsa-miR-96) were overexpressed, whereas 84 miRNAs (hsa-miR-1, hsa-miR-145, hsa-miR-145*, and so on) were downexpressed in tumor tissues compared with those in normal tissues.

10

Six miRNAs of them (miR-450b-5p, miR-424, miR-503, miR-542-3p, miR-629, and miR-214) were significantly underexpressed, while one miRNA (miR-592) was significantly overexpressed in NFA compared to normal pituitary tissues.

11

In another report, miR-592 promotes metastasis, in part, by targeting FOXO3A in human colorectal cancer [79].

12

10 miRNAs were consistently validated (Fig.   6) to be differentially expressed by HP diet, namely miR-592, miR-488-3p, miR-339-3p, miR-17-5p-1, miR-877-5p, miR-15b-5p, miR-484-1, miR-4745-5p, let-7b-5p and miR-25-3p.

13

In the cerebellum, novel_circRNA_007362 was predicted to combine with 24 miRNAs (tch-let-7e-5p, tch-let-7i-5p, tch-let-7f-5p, tch-miR-125a-5p, tch-miR-1301, tch-miR-135a-5p, tch-miR-135b-5p, tch-miR-15b-5p, tch-miR-195-5p, tch-miR-1-5p, tch-miR-218-5p, tch-miR-22-3p, tch-miR-26a-5p, tch-miR-26b-5p, tch-miR-296-3p, tch-miR-335-5p, tch-miR-34a-5p, tch-miR424-5p,tch-miR-491-5p, tch-miR-455-3p,tch-miR-503, tch-miR-592, tch-miR-9771e, and tch-miR-98-5p).

14

Other miRNAs controlling both proliferation and differentiation of adult NSPCs are miR-137 [122] and rno-miR-592 [123].