DUSP1 induces paclitaxel resistance through the regulation of p-glycoprotein expression in human ovarian cancer cells

Kang, Yu-Seon,Seok, Hyun-Jeong,Jeong, Eun-Jeong,Kim, Yuna,Yun, Seok-Joong,Min, Jeong-Ki,Kim, Sun Jin,Kim, Jang-Seong Elsevier 2016 Biochemical and biophysical research communication Vol.478 No.1

<P><B>Abstract</B></P> <P>The heterogeneity and genetic instability of ovarian cancer cells often lead to the development of drug resistance, closely related with the increased cancer-related mortality. In this study, we investigated the role of dual-specificity phosphatase 1 (DUSP1) in the development of the resistance in human ovarian cancer cells against paclitaxel. Overexpression of DUSP1 in HeyA8 human ovarian cancer cells (HeyA8-DUSP1) up-regulated the expression of the drug efflux pump, p-glycoprotein. Consequently, HeyA8-DUSP1 cells are highly resistant to paclitaxel, with the resistance comparable to that of a multi-drug resistance cell line (HeyA8-MDR). Moreover, over expression of DUSP1 significantly increased the activation of p38 MAPK, leaving the activation of ERK1/2 and JNK1/2 unaffected. Pharmacological suppression of p38 MAPK activity prevents the up-regulation of p-glycoprotein expression and the consequent resistance against paclitaxel in HeyA8-DUSP1 cells. By contrast, HeyA8-MDR cells expressed a significantly higher level of DUSP1, but treatment with small interference RNA against DUSP1 significantly suppressed the expression of p-glycoprotein and the resistance against paclitaxel in HeyA8-MDR cells. Ectopic expression of MKK3, an upstream activator of p38 MAPK, significantly up-regulated the expression of p-glycoprotein and increased the consequent resistance against paclitaxel in HeyA8 cells. Collectively, these data indicated that DUSP1 may induce the resistance against paclitaxel through the p38 MAPK-mediated overexpression of p-glycoprotein in human ovarian cancer cells.</P> <P><B>Highlights</B></P> <P> <UL> <LI> DUSP1 induces Pgp expression and paclitaxel resistance in ovarian cancer cells. </LI> <LI> DUSP1 mediates p38 MAPK activation leading to Pgp-mediated paclitaxel resistance. </LI> <LI> Inhibition of DUSP1 or p38 MAPK reverses Pgp-mediated paclitaxel resistance. </LI> <LI> DUSP1-p38 MAPK-Pgp axis is a novel mechanism for paclitaxel resistance. </LI> </UL> </P>

Basic, HCCbasic : PO-20 ; DUSP1 induces p53 target gene expression through p38MAPK/HSP27 pathway and tumor suppression in hepatocellular carcinoma

( Pei Pei Hao ),( Mi Jin Lee ),( Yun Peng Wang ),( Goung Ran Yu ),( In Hee Kim ),( Dae Ghon Kim ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1

Background: Constitutive DUSP1 expression has been shown to be involved in cell cycle inhibition, apoptosis, and senescence. This study was aimed to examine whether DUSP1 functions as tumor suppressor in hepatocarcinogenesis and to explore underlying mechanism whereby DUSP1 suppresses hepatocarcinogenesis. Methods: Immunohistochemistry and real-time PCR analysis were performed in HCC tissues. Cellular localization of DUSP1 was detected by immunofluorescence. Cell proliferation was tested by MTT assay. Cell death and cell cycle were measured by FACS analysis. Apoptotic and kinase signaling were explored by western blot analysis. Tumorigenicity and survival analysis were tested by xenotransplant of SH-J1 cells stably expressing DUSP1 or infected with Ad-DUSP1 in mouse model. Phospho-related factors expression profile in DUSP1 stable cell lines as determined by phospho-kinase array. Results: The mRNA and protein expression level of DUSP1 was down-regulated in tumor than that of the corresponding non-tumor in HCC tissues. Cellular localization of DUSP1 showed that the endogenous DUSP1 and ectopic expression of GFP-tagged DUSP1 was mainly located in the nucleus. DUSP1 down-regulation was associated with reciprocal activation of ERK1/2 in HCC cell lines. DUSP1 was up-regulated in a dose dependent manner after parthenolide or doxorubicin treatment. DUSP1 over-expression was correlated with the increased susceptibility to apoptotic cell death through caspase activation. Ectopic DUSP1 over-expression resulted in the inhibitions of cell cycle progression, colony generation, and tumor growth in vitro and in vivo system. Furthermore, survival rate of mice xenoplanted with DUSP1 overexpressed HCC cells is significantly higher than control group. Inhibition of tumorigenic potential by DUSP1 may involve in p38MAPK- HSP27-P53 pathway. Conclusions: DUSP1 functions as a tumor suppressor during hepatocarcinogenesis, which seemed to be mainly associated with the activation of p53 target genes through p38MAPK/ HSP27 pathway.

Disruption of a regulatory loop between DUSP1 and p53 contributes to hepatocellular carcinoma development and progression

Hao, P.P.,Li, H.,Lee, M.J.,Wang, Y.P.,Kim, J.H.,Yu, G.R.,Lee, S.Y.,Leem, S.H.,Jang, K.Y.,Kim, D.G. Elsevier Science Publishers 2015 Journal of hepatology Vol.62 No.6

Background & Aims: Altered expression of dual specificity phosphatase 1 (DUSP1) is common in tumors including hepatocellular carcinoma (HCC), and is predictive of tumor progression and poor prognosis. However, the tumor suppressive role of DUSP1 has yet to be clearly elucidated. Methods: The molecular mechanisms of tumor suppression that were investigated were induction of apoptosis, cell cycle inhibition, and regulation of p53. Additionally, the antitumor effect of DUSP1 was assessed using a mouse model. Associated signaling pathways in HCC cells and tissues were examined. Results: Downregulation of DUSP1 expression was significantly correlated with poor differentiation (p<0.001) and advanced HCC stage (p=0.023). DUSP1 expression resulted in HCC suppression and longer survival (p=0.0002) in a xenoplant mice model. DUSP1 inhibited p38 MAPK phosphorylation and subsequently suppressed HSP27 activation, resulting in enhanced p53 phosphorylation at sites S15, S20, and S46 in HCC cells. Enhanced p53 activation induced the expression of target genes p21 and p27, which are linked to cell cycle arrest and apoptosis. Thus, DUSP1 was potentially linked to p53 activation via the p38 MAPK/HSP27 pathway. Wild-type but not mutant p53 transcriptionally upregulated DUSP1 via its DNA-binding domain. DUSP1 and p53 might collaborate to suppress tumors in hepatocarcinogenesis via a positive regulatory loop. Conclusions: Our results revealed that disruption of a positive regulatory loop between DUSP1 and p53 promoted HCC development and progression, providing a rationale for a therapeutic agent that restores DUSP1 in HCC.

Cks1 regulates human hepatocellular carcinoma cell progression through osteopontin expression

Kang, Yu-Seon,Jeong, Eun-Jeong,Seok, Hyun-Jeong,Kim, Seon-Kyu,Hwang, Jin-Seong,Choi, Mu Lim,Jo, Dong-Gyu,Kim, Yuna,Choi, Jinhyeon,Lee, Yeo-Jin,Jung, Eunsun,Min, Jeong-Ki,Han, Tae-Su,Kim, Jang-Seong Elsevier 2019 Biochemical and biophysical research communication Vol.508 No.1

<P><B>Abstract</B></P> <P>Precise cell cycle regulation is critical to prevent aberrant cell proliferation and cancer progression. Cks1 was reported to be an essential accessory factor for SCF<SUP>Skp2</SUP>, the ubiquitin ligase that targets p27<SUP>Kip1</SUP> for proteasomal degradation; these actions drive mammalian cell transition from G1 to S phase. In this study, we investigated the role played by Cks1 in the growth and progression of human hepatocellular carcinoma (HCC) cells. Silencing Cks1 expression abrogated osteopontin (OPN) expression in a p27<SUP>Kip1</SUP>-dependent manner in Huh7 HCC cells. OPN increased the proliferation, migration and invasion of Huh7 cells. Pharmacological inhibitor studies demonstrated that ERK1/2 signaling is responsible mainly for Cks1-mediated OPN expression. Cks1 appears to regulate ERK1/2 signaling through the expression of dual-specificity phosphatase 16 (DUSP16) because both Cks1 knockdown, which leads to DUSP16 upregulation, and DUSP16 overexpression decreased ERK1/2 phosphorylation and the resulting OPN expression. The same is true for the Cks1-mediated increases in p27<SUP>Kip1</SUP>, suggesting that Cks1 regulates OPN expression through activating ERK1/2 signaling either by suppressing DUSP16 expression or by a p27<SUP>Kip1</SUP>-dependent mechanism. Cks1 and OPN expression levels were significantly higher, but DUSP16 expression levels were significantly lower in HCC tissues than in normal liver tissues. Both Cks1 and OPN expression were negatively correlated with DUSP16 expression, whereas Cks1 expression was positively correlated with OPN expression. Moreover, combined panels for the expression levels of Cks1, DUSP16 and OPN showed significant prognostic power for the risk assessment of HCC patient overall survival. In conclusion, our data propose a novel function for Cks1 as a tumor promoter through the expression of the strongly oncogenic protein OPN in HCC.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Cks1 promotes OPN expression in human HCC cells. </LI> <LI> The Cks1-mediated upregulation of DUSP16 or p27<SUP>Kip1</SUP> decreased OPN expression. </LI> <LI> DUSP16- or p27<SUP>Kip1</SUP>-mediated ERK1/2 inactivation is responsible for OPN abrogation. </LI> <LI> Cks1, DUSP16 and OPN may be a useful prognostic biomarker panel for HCC patients. </LI> </UL> </P>

Calcium-Phosphate Crystals Promote RANKL Expression via the Downregulation of DUSP1

Choi, YunJeong,Yoo, Ji Hyun,Lee, Youngkyun,Bae, Moon Kyoung,Kim, Hyung Joon Korean Society for Molecular and Cellular Biology 2019 Molecules and cells Vol.42 No.2

Osteoarthritis (OA) is a naturally occurring, irreversible disorder and a major health burden. The disease is multifactorial, involving both physiological and mechanical processes, but calcium crystals have been associated intimately with its pathogenesis. This study tested the hypothesis that these crystals have a detrimental effect on the differentiation of osteoclasts and bone homeostasis. This study employed an osteoblastosteoclast coculture system that resembles in vivo osteoblastdependent osteoclast differentiation along with $Ca^{2+}$-phosphate-coated culture dishes. The calcium-containing crystals upregulated the expression of RANKL and increased the differentiation of osteoclasts significantly as a result. On the other hand, osteoblast differentiation was unaffected. MicroRNA profiling showed that dual-specificity phosphatases 1 (DUSP1) was associated with the increased RANKL expression. DUSP1 belongs to a family of MAPK phosphatases and is known to inactivate all three groups of MAPKs, p38, JNK, and ERK. Furthermore, knockdown of DUSP1 gene expression suggested that RANKL expression increases significantly in the absence of DUSP1 regulation. Microarray analysis of the DUSP1 mRNA levels in patients with pathological bone diseases also showed that the downregulated DUSP1 expression leads to increased expression of RANKL and consequently to the destruction of the bone observed in these patients. These findings suggest that calcium-containing crystals may play a crucial role in promoting RANKL-induced osteoclastogenesis via DUSP1.

Calcium-Phosphate Crystals Promote RANKL Expression via the Downregulation of DUSP1

최윤정,유지현,이영균,배문경,김형준 한국분자세포생물학회 2019 Molecules and cells Vol.42 No.2

Osteoarthritis (OA) is a naturally occurring, irreversible disorder and a major health burden. The disease is multifactorial, involving both physiological and mechanical processes, but calcium crystals have been associated intimately with its pathogenesis. This study tested the hypothesis that these crystals have a detrimental effect on the differentiation of osteoclasts and bone homeostasis. This study employed an osteoblast-osteoclast coculture system that resembles in vivo osteoblast-dependent osteoclast differentiation along with Ca2+-phosphate-coated culture dishes. The calcium-containing crystals upregulated the expression of RANKL and increased the differentiation of osteoclasts significantly as a result. On the other hand, osteoblast differentiation was unaffected. MicroRNA profiling showed that dual-specificity phosphatases 1 (DUSP1) was associated with the increased RANKL expression. DUSP1 belongs to a family of MAPK phosphatases and is known to inactivate all three groups of MAPKs, p38, JNK, and ERK. Furthermore, knockdown of DUSP1 gene expression suggested that RANKL expression increases significantly in the absence of DUSP1 regulation. Microarray analysis of the DUSP1 mRNA levels in patients with pathological bone diseases also showed that the downregulated DUSP1 expression leads to increased expression of RANKL and consequently to the destruction of the bone observed in these patients. These findings suggest that calcium-containing crystals may play a crucial role in promoting RANKL-induced osteoclastogenesis via DUSP1.

Dusp1 modulates activin/smad2 mediated germ layer specification via FGF signal inhibition in Xenopus embryos

Zobia Umair,Santosh Kumar,Khezina Rafiq,Vijay Kumar,Zia Ur Reman,이승환,김성찬,이재용,이은주,김재봉 한국통합생물학회 2020 Animal cells and systems Vol.24 No.6

Activin, a member of the transforming growth factor (TGF-β) superfamily, induces mesoderm, endoderm and neuro-ectoderm formation in Xenopus embryos. Despite several previous studies, the complicated gene regulatory network and genes involved in this induction await more elaboration. We identified expression of various fibroblast growth factor (FGF) genes in activin/ smad2 treated animal cap explants (AC) of Xenopus embryos. Activin/smad2 increased fgf3/8 expression, which was reduced by co-injection of dominant negative activin receptor (DNAR) and dominant negative Fgf receptor (DNFR). Interestingly, activin/smad2 also increased expression of dual specificity phosphatase 1 (dusp1) which has been known to inhibit Fgf signaling. Dusp1 overexpression in dorsal marginal zone caused gastrulation defect and decreased Jnk/Erk phosphorylation as well as Smad1 linker region phosphorylation. Dusp1 decreased neural and organizer gene expression with increasing of endodermal and ventral gene expression in smad2 treated AC, indicating that dusp1 modulates germ layer specification. Dusp1 decreased neural gene expression in fgf8 treated AC, suggesting that Erk and/or Jnk phosphorylation may be involved in fgf8 induced neural induction. In addition, dusp1 decreased the reporter gene activities of activin response element (ARE) and increased it for bmp response element (BRE), indicating that dusp1 modulates two opposite morphogen signaling of dorsal (activin/Smad2) and ventral (bmp/Smad1) tracks, acting to fine tune the Fgf/Erk pathway.

Comprehensive Analysis of Vascular Endothelial Growth Factor-C Related Factors in Stomach Cancer

Liu, Yong-Chao,Zhao, Jing,Hu, Cheng-En,Gan, Jun,Zhang, Wen-Hong,Huang, Guang-Jian Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.5

Background: Vascular endothelial growth factor-C (VEGF-C), which contributes to lymphatic metastasis (LM) in malignant disease, is one of the most important factors involved in physical and pathological lymphangiogenesis. Some VEGF-C related factors such as sine oculis homeobox homolog (SIX) 1, contactin (CNTN) 1 and dual specificity phosphatase (DUSP) 6 have been extensively studied in malignancies, but their expression levels and associations have still to be elucidated in stomach cancer. Methods: We detected their expression levels in 30 paired stomach cancer tissues using quantitative real-time reverse transcription-PCR (qRT-PCR). The expression and clinical significance of each factor was analyzed using Wilcoxon signed rank sum test. The correlation among all the factors was performed by Spearman rank correlation analysis. Results: The results suggest that VEGF-C and CNTN1 are significantly correlated with tumor size, SIX1 with the age and CNTN1 also with the cTNM stage. There are significant correlations of expression levels among VEGF-C, SIX1, CNTN1 and DUSP6. Conclusions: There exists an important regulatory crosstalk involving SIX1, VEGF-C, CNTN1 and DUSP6 in stomach cancer.

The mitogen-activated protein kinase phosphatase-1 (MKP-1) gene is a potential methylation biomarker for malignancy of breast cancer

Fang-Ming Chen,Hsueh-Wei Chang,Sheau-Fang Yang,Ya-Fang Huang,Pei-Yung Nien,Yao-Tsung Yeh,Ming-Feng Hou 생화학분자생물학회 2012 Experimental and molecular medicine Vol.44 No.5

The mitogen-activated protein kinase (MAPK) phosphatase-1 (MKP-1) belongs to the MAPK cascades which are central to cell proliferation and apoptosis. The carcinogenic role of MKP-1 has been reported in many types of cancer but it has rarely been investigated in breast cancer. The present study was designed to evaluate the MKP-1 mRNA expression and its possible regulation by methylation of MKP-1 promoter in the model of several breast cancer cell lines and tissues as well as controls. Our data demonstrate MKP-1mRNA expression significantly decreased in five breast cancer cell lines compared to breast controls (P <0.01). Using the methylation-specific PCR (MSP) analysis,the unmethylated reaction (U) is dominant in both normal cell lines and benign breast tumors (100% vs. 86.2%), whereas the methylated reaction (M) is dominant in both breast cancer cell lines and invasive breast tumors (100% vs. 57.2%). In terms of methylation ratio (M/M+U), methylation level in MKP-1 promoter is significantly higher in the invasive breast tumor tissues (n = 152) than in benign breast tumor tissues (n = 29)(P < 0.0001). Assessing the methylation ratio of the promoter of MKP-1 gene to diagnose the breast malignancy (invasive vs. benign), the area under the receiver-operating characteristic (ROC) curve was 0.809(95% CI: 0.711-0.906, P < 0.001). The best performance for this prediction has a sensitivity of 76.32% and a specificity of 82.76% at the cutoff value of 0.38. Taken together, we firstly demonstrated that the promoter methylation of MKP-1 gene is a potential breast cancer biomarker for breast malignancy.

TSH Signaling Overcomes B-RafV600E–Induced Senescence in Papillary Thyroid Carcinogenesis through Regulation of DUSP6 ¹ ²

Kim, Young Hwa,Choi, Yong Won,Han, Jae Ho,Lee, Jeonghun,Soh, Euy Young,Park, So Hyun,Kim, Jang-Hee,Park, Tae Jun Neoplasia Press 2014 Neoplasia Vol.16 No.12

<P>B-RafV600E oncogene mutation occurs most commonly in papillary thyroid carcinoma (PTC) and is associated with tumor initiation. However, a genetic modification by B-RafV600E in thyrocytes results in oncogene-induced senescence (OIS). In the present study, we explored the factors involved in the senescence overcome program in PTC. First of all, we observed down-regulation of p-extracellular signal-regulated kinases 1/2 and up-regulation of dual specific phosphatase 6 (DUSP6) in the PTC with B-RafV600E mutation. DUSP6 overexpression <I>in vitro</I> induced extracellular signal-regulated kinases 1/2 dephosphorylation and inhibited B-RafV600E–induced senescence in thyrocytes. Although DUSP6 protein was degraded by B-RafV600E–induced reactive oxygen species (ROS), thyroid-stimulating hormone (TSH) stabilized DUSP6 protein by increasing Mn superoxide dismutase expression and inhibited B-RafV600E–induced senescence. Although serum TSH was not increased, its receptor was markedly upregulated in PTC with B-RafV600E. Furthermore, TSH together with DUSP6 reactivated Ras signaling, resulted in activation of Ras/AKT/glycogen synthase kinase 3β, and stabilized c-Myc protein by inhibiting its degradation. These observations led us to conclude that increased TSH signaling overcomes OIS and is essential for B-RafV600E–induced papillary thyroid carcinogenesis.</P>