Decreased Expression of the Suppressor of Cytokine Signaling 6 in Human Hepatocellular Carcinoma

Bae, Hyun-Jin,Noh, Ji-Heon,Eun, Jung-Woo,Kim, Jeong-Kyu,Jung, Kwang-Hwa,Xie, Hong Jian,Ahn, Young-Min,Ryu, Jae-Chun,Park, Won-Sang,Lee, Jung-Young,Nam, Suk-Woo The Korean Society of Toxicogenomics and Toxicopro 2009 Molecular & cellular toxicology Vol.5 No.3

Suppressors of cytokine signaling (SOCS) proteins were originally identified as negative feedback regulators of cytokine signaling and include the Janus kinase/Signal transducer and activator of transcription (JAK/STAT) pathways. Recent studies have shown that SOCS proteins negatively regulate the receptor tyrosine kinase (RTK) pathway including the insulin receptor (IR), EGFR, and KIT signaling pathways. In addition, SOCS1 and SOCS3 have been reported to have anti-tumor effects in human hepatocellular carcinoma (HCC). However, it is uncertain whether other members of the SOCS family are associated with tumor development and progression. In this study, to investigate whether SOCS6 is aberrantly regulated in HCC, we examined the expression level of SOCS6 in HCC by Western blot analysis and immunohistochemical staining. The results showed that SOCS6 was down-regulated in all examined HCCs compared to the corresponding normal tissues. In addition, expression of SOCS6 was observed in the cytoplasm of most normal and precancerous tissue, but not in the HCCs by immunohistochemical staining. This is first report to demonstrate that SOCS6 is aberrantly regulated in HCC. These findings suggest that underexpression of SOCS6 is involved in hepatocarcinogenesis, and SOCS6 may play a role, as a tumor suppressor, in HCC development and progression.

Analysis on SOCS-Antigen Protein correlated with Prognostic Factors of Oral Cancer Patients

유미현,장민아,한혜연,김욱규 대한구강악안면병리학회 2023 대한구강악안면병리학회지 Vol.47 No.3

SOCS3, a suppressor of cytokine signaling 3, is known as a negative regulator of various cytokines and a tumor suppressor gene in human tumors. This study aimed to investigate the role of SOCS3 in oral squamous cell carcinoma (OSCC) and its impact on epithelial-mesenchymal transition (EMT) in OSCC cells. Although SOCS3 is recognized as a negative regulator of various cytokines and a tumor suppressor gene in human tumors, its specific effects on OSCC remain poorly understood. For the assessment of SOCS3 expression in OSCC, the UALCAN website and TCGA data were used to evaluate its expression in head and neck cancer. Additionally, immunohistochemical staining was conducted to determine the SOCS3 expression specifically in OSCC. The findings indicated a significant decrease in SOCS3 expression in tumor tissue compared to that in normal tissues. To investigate the enhancement of SOCS3 expression in OSCC cancer cell lines, IL6 treatment was administered to MC3 cells. However, no significant differences were observed in cell viability, wound healing assay, and invasion assay. Conversely, the transfection of SOCS3 siRNA into OSCC cells led to a notable increase in cell viability and statistically significant increases in wound healing and invasion assays. These results suggest that SOCS3 plays a crucial role in cell viability and EMT in OSCC, thereby contributing to oral carcinogenesis. Further research is necessary to elucidate the precise role of SOCS3 in OSCC.

Increased cytoplasmic levels of CIS, SOCS1, SOCS2, or SOCS3 are required for nuclear translocation

Lee, Kyeong-Hee,Moon, Kyeong-Jin,Kim, Hyung Sik,Yoo, Byong Chul,Park, Seoyoung,Lee, Ho,Kwon, Soim,Lee, Eun Sook,Yoon, Sungpil Elsevier 2008 FEBS letters Vol.582 No.15

<P><B>Abstract</B></P><P>We investigated the cellular localization of ectopically-expressed CIS, SOCS1, SOCS2 and SOCS3 proteins. We found that SOCS proteins localize to the nucleus where they reduce Stat3 proteins and that the presence of proteasome inhibitors increased SOCS nuclear localization. Our results indicate that increased nuclear localization resulted from increased levels of SOCS proteins in the cytoplasm. Finally, we demonstrate that the same effect occurs with endogenously-expressed SOCS proteins. These observations suggest that increased cytoplasmic levels of proteins in the SOCS family are regulated through nuclear translocation.</P>

Down-regulation of microRNA-155 suppressed Candida albicans induced acute lung injury by activating SOCS1 and inhibiting inflammation response

Li Xiaohua,Gong Yuanzhong,Lin Xin,Lin Qiong,Luo Jianxiong,Yu Tianxing,Xu Junping,Chen Lifang,Xu Liyu,Hu Ying 한국미생물학회 2022 The journal of microbiology Vol.60 No.4

Acute lung injury caused by Candida albicans could result in high mortality and morbidity. MicroRNA-155 (miR-155) and suppressor of cytokine signaling 1 (SOCS1) have been believed to play a key in the regulation of inflammatory response. Whether miR-155/SOCS1 axis could regulate the acute lung injury caused by C. albicans has not been reported. The acute lung injury animal model was established with acute infection of C. albicans. miR-155 inhibitor, miR-155 mimic, and sh-SOCS1 were constructed. The binding site between miR- 155 and SOCS1 was identified with dual luciferase reporter assay. Knockdown of miR-155 markedly inhibited the germ tube formation of C. albicans. Knockdown of miR-155 significantly up-regulated the expression of SOCS1, and the binding site between miR-155 and SOCS1 was identified. Knockdown of miR-155 improved the acute lung injury, suppressed inflammatory factors and fungus loading through SOCS1. Knockdown of SOCS1 greatly reversed the influence of miR- 155 inhibitor on the cell apoptosis in vitro. The improvement of acute lung injury caused by C. albicans, suppression of inflammatory response and C. albicans infection, and inhibitor of cell apoptosis were achieved by knocking down miR-155 through SOCS1. This research might provide a new thought for the prevention and treatment of acute lung injury caused by C. albicans through targeting miR-155/SOCS1 axis.

Anti-inflammatory mechanisms of suppressors of cytokine signaling target ROS via NRF-2/thioredoxin induction and inflammasome activation in macrophages

( Ga-young Kim ),( Hana Jeong ),( Hye-young Yoon ),( Hye-min Yoo ),( Jae Young Lee ),( Seok Hee Park ),( Choong-eun Lee ) 생화학분자생물학회 2020 BMB Reports Vol.53 No.12

Suppressors of cytokine signaling (SOCS) exhibit diverse antiinflammatory effects. Since ROS acts as a critical mediator of inflammation, we have investigated the anti-inflammatory mechanisms of SOCS via ROS regulation in monocytic/macrophagic cells. Using PMA-differentiated monocytic cell lines and primary BMDMs transduced with SOCS1 or shSOCS1, the LPS/TLR4- induced inflammatory signaling was investigated by analyzing the levels of intracellular ROS, antioxidant factors, inflammasome activation, and pro-inflammatory cytokines. The levels of LPS-induced ROS and the production of pro-inflammatory cytokines were notably down-regulated by SOCS1 and up-regulated by shSOCS1 in an NAC-sensitive manner. SOCS1 up-regulated an ROS-scavenging protein, thioredoxin, via enhanced expression and binding of NRF-2 to the thioredoxin promoter. SOCS3 exhibited similar effects on NRF-2/thioredoxin induction, and ROS downregulation, resulting in the suppression of inflammatory cytokines. Notably thioredoxin ablation promoted NLRP3 inflammasome activation and restored the SOCS1-mediated inhibition of ROS and cytokine synthesis induced by LPS. The results demonstrate that the anti-inflammatory mechanisms of SOCS1 and SOCS3 in macrophages are mediated via NRF-2-mediated thioredoxin upregulation resulting in the downregulation of ROS signal. Thus, our study supports the anti-oxidant role of SOCS1 and SOCS3 in the exquisite regulation of macrophage activation under oxidative stress. [BMB Reports 2020; 53(12): 640-645]

노화 흰쥐 전경골근 및 해마의 SOCS3 및 IL15 단백질 발현에 대한 운동트레이닝 효과

안나영(AhnNaYoung),김기진(KimKiJin),임창현(LimChangHyun),김창근(KimChangKeun) 한국체육학회 2018 한국체육학회지 Vol.57 No.2

본 연구는 노화과정에서 염증발현 증가에 의한 퇴행성 뇌질환을 예방하기 위하여 운동 유형에 따른 항 염증성 사이토카인 발현 효과를 골격근 및 해마조직에서 분석하였다. 연구대상은 50주령 Wistar계 수컷 흰쥐 40마리를 대조군(Con; n=10), 유산소운동군(AT; n=10), 저항성 운동군(RT; n=10), 복합 운동군(AT+RT; n=10)으로 구분하였다. 유산소성 운동인 트레드밀운동과 저항성 운동형태인 사다리 등반운동을 적응시켜 12주간 그룹에 따른 처치를 실시하였다. 흰쥐의 전경골근 및 해마를 적출하여 TLR4, TNF-α, SOCS3, IL-15 단백질 발현을 분석하였다. 본 연구 결과 모든 운동군의 체중은 대조군에 비해 유의하게 (p<.05) 감소하였다. 복합운동군의 전경골근의 SOCS3 단백질 발현은 대조군 및 저항성 운동군에 비해서 유의하게(p<.05) 높게 나타났으며, SOCS3/TLR4도 유산소 운동군에 비해 높게(p=0.054) 나타났다. 해마의 SOCS3 단백질 발현은 모든 운동군에서 대조군에 비해 유의하게(p<.05) 증가하였다. 해마의 IL-15 단백질 발현도 저항성 운동 및 복합운동군이 대조군에 비해 유의하게(p<.05) 증가하였다. 그러므로 노화 흰쥐의 12주간 운동은 전경골근과 해마에서 SOCS3 및 IL-15의 발현이 현저하게 증가하여 항 염증성 사이토카인의 증가로 퇴행성 뇌질환 예방에 긍정적인 영향을 미칠 것으로 생각된다. The purpose of this study was to investigate the effect of anti-inflammatory cytokine expression on skeletal muscle and hippocampus to protect degenerative brain diseases caused by increased inflammation during aging process. The study groups were consisted of 40 Wistar old male rats at 50 weeks, and were divided into 4 groups as a control group (Con; n=10), an aerobic training group (AT; n=10), a resistance training group (RT; n=10), and combined training group (AT+RT; n=10). Exercise training program included of ladder-climbing exercise and treadmill exercise, 3 days per week, for 12 weeks. The expression of TLR4, TNF-α, SOCS3 and IL-15 protein was analyzed by western blot. The body weight of all exercise groups was significantly (p<.05) lower than that of the control group. SOCS3 protein expression in the combine exercise group was significantly (p<.05) higher than the control and resistance exercise group, and SOCS3/TLR4 in the combine exercise group was also higher than the aerobic exercise group (p=0.054). The expression of SOCS3 protein in hippocampus was significantly (p<.05) increased in all exercise groups compared to the control group. IL-15 protein expression in the hippocampus was increased in the resistance exercise group (p<.05) and combine exercise group (p<.05) compared to the control group. Therefore, the 12-week exercise training in aged rats significantly increase the expression of SOCS3 and IL15 in both skeletal muscle and hippocampus, resulting positive effect on degenerative brain diseases by increasing the anti-inflammatory cytokines.

Anti-inflammatory actions of probiotics through activating suppressor of cytokine signaling (SOCS) expression and signaling in <i>Helicobacter pylori</i> infection: A novel mechanism

Lee, Jeong Sang,Paek, Nam Soo,Kwon, Oh Sang,Hahm, Ki Baik Blackwell Publishing Asia 2010 Journal of gastroenterology and hepatology Vol.25 No.1

<P>Abstract</P><P>Background and Aims: </P><P>In spite of the International Agency for Research on Cancer's definition that <I>Helicobacter pylori</I> is the definite carcinogen of gastric cancer, the simple eradication of the bug is not enough to prevent resultant gastric cancer, and increasing microbial resistance further limits the eradication application. Therefore, probiotics, non-pathogenic microbial feed that can affect the host in a beneficial manner, could be an alternate way to enhance anti-inflammation against <I>H. pylori</I>. However, the mechanism of their anti-inflammatory actions is still unclear. In the current study, we hypothesized that suppressor of cytokine signaling (SOCS) signaling could be a feasible anti-inflammatory mechanism of probiotics against <I>H. pylori</I> infection.</P><P>Results: </P><P><I>H. pylori</I> infection or their lipopolysaccharide stimulation led to significant increased expressions of inflammatory mediators including tumor necrosis factor-&agr;, interleukin-8, inducible nitric oxide synthase and cyclooxygenase-2 in AGS cells and pretreatment of <I>Lactobacillus plantarum</I>, <I>Lactobacillus rhamnosis</I> and <I>Lactobacillus acidophilus</I> significantly attenuated the expressions of these inflammatory mediators in accordance with the blocking action of nuclear factor-&kgr;B nuclear translocation. Probiotic administration increased expression of SOCS-2 and SOCS-3 and exerted the active SOCS signaling featured with earlier and higher expressions of SOCS-2 and SOCS-3. In contrast to weak inactivation of mitogen-activated protein kinases including p-38 and extracellular signal-regulated kinase 1/2, probiotic-induced SOCS expressions were mediated through either significant phosphorylation of signal transducers and activation of transcription (STAT)-1 and STAT-3 or simultaneous inhibition of Janus kinase (JAK)2 phosphorylation, which is known to signal SOCS-2/SOCS-3 negatively.</P><P>Conclusion: </P><P>Anti-inflammatory signals of SOCS through STAT-1/STAT-3 activation and JAK2 inactivation might be a key anti-inflammatory mechanism of probiotics, setting probiotics as a non-microbial strategy to <I>H. pylori</I> infection.</P>

SOCS1 and SOCS3 are expressed in mononuclear cells in human cytomegalovirus viremia after allogeneic hematopoietic stem cell transplantation

신승환,이지윤,윤재호,이태향,박소혜,양승아,김희제,이성은,조병식,이동건,김유진,이석,민창기,조석구,김동욱,이종욱,민우성,박종원 대한혈액학회 2015 Blood Research Vol.50 No.1

BackgroundThe expression of the SOCS genes in cytomegalovirus (CMV) viremia after hematopoieticstem cell transplantation (HSCT) remains largely unexplored. MethodsUsing quantitative RT-PCR of mononuclear cells, we conducted pairwise comparison ofSOCS1 and SOCS3 expression levels among a healthy donor group (N=55), a pre-HSCTgroup (N=17), and the recipient subgroup (N=107), which were divided according tothe occurrence of CMV viremia and acute graft-versus-host disease (aGVHD). ResultsCompared to that in the healthy donor group, SOCS1 expression was higher in the CMV+subgroup, especially in the CMV+GVHD- group, but decreased in the other subgroups. When compared to the expression in the pre-HSCT group, SOCS1 expression was significantlyhigher in the CMV+ subgroup, especially in the CMV+GVHD+ subgroup. Meanwhile, compared to that in the healthy donor group, SOCS3 expression was significantlylower in all other groups. The CMV-GVHD- subgroup showed significantly lowerSOCS3 expression compared to the CMV+ subgroup, the CMV+GVHD+ subgroup,and the CMV+GVHD- subgroup. ConclusionWe report differential expression of SOCS genes according to CMV viremia with acuteGVHD occurrence after HSCT, suggesting that regulation of SOCS expression is associatedwith CMV viremia.

SOCS3 suppresses the expression of IL-4 cytokine by inhibiting the phosphorylation of c-Jun through the ERK signaling pathway in rat mast cell line RBL-2H3

Kim, D.,Kim, S.H.,Cho, S.H.,Shin, K.,Kim, S. Pergamon Press 2011 Molecular immunology Vol.48 No.5

SOCS3 is well known to negatively regulate various cytokine-mediated signaling responses, but its direct role in the expression of specific cytokines has not been clearly elucidated. To understand the role of SOCS3 in the expression of IL-4, one of the key Th2 cytokines, RBL-2H3 cells (a rat mast cell line) were engineered to express SOCS3 constitutively at a high level or at a lower level using shRNA. In RBL-2H3 cells stably expressing SOCS3, the RNA and protein levels of IL-4 were significantly decreased, while it was opposite in RBL-2H3 cells containing shRNA for SOCS3. Overexpression of SOCS3 was found to reduce the level of calcium ionophore-induced phosphorylation of ERK½ and c-Jun transcription factor. Consistent with this data, knockdown of SOCS3 increased the level of phosphorylated ERK1 and ERK2. Taken together, SOCS3 appears to play an important role as a negative feedback inhibitor in the expression of IL-4 by inhibiting serine phosphorylation of c-Jun via the ERK signaling pathway.

Silencing of Suppressor of Cytokine Signaling-3 due to Methylation Results in Phosphorylation of STAT3 in Imatinib Resistant BCR-ABL Positive Chronic Myeloid Leukemia Cells

Al-Jamal, Hamid AN,Jusoh, Siti Asmaa Mat,Yong, Ang Cheng,Asan, Jamaruddin Mat,Hassan, Rosline,Johan, Muhammad Farid Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.11

Background: Silencing due to methylation of suppressor of cytokine signaling-3 (SOCS-3), a negative regulator gene for the JAK/STAT signaling pathway has been reported to play important roles in leukemogenesis. Imatinib mesylate is a tyrosine kinase inhibitor that specifically targets the BCR-ABL protein and induces hematological remission in patients with chronic myeloid leukemia (CML). Unfortunately, the majority of CML patients treated with imatinib develop resistance under prolonged therapy. We here investigated the methylation profile of SOCS-3 gene and its downstream effects in a BCR-ABL positive CML cells resistant to imatinib. Materials and Methods: BCR-ABL positive CML cells resistant to imatinib (K562-R) were developed by overexposure of K562 cell lines to the drug. Cytotoxicity was determined by MTS assays and $IC_{50}$ values calculated. Apoptosis assays were performed using annexin V-FITC binding assays and analyzed by flow cytometry. Methylation profiles were investigated using methylation specific PCR and sequencing analysis of SOCS-1 and SOCS-3 genes. Gene expression was assessed by quantitative real-time PCR, and protein expression and phosphorylation of STAT1, 2 and 3 were examined by Western blotting. Results: The $IC_{50}$ for imatinib on K562 was 362nM compared to 3,952nM for K562-R (p=0.001). Percentage of apoptotic cells in K562 increased upto 50% by increasing the concentration of imatinib, in contrast to only 20% in K562-R (p<0.001). A change from non-methylation of the SOCS-3 gene in K562 to complete methylation in K562-R was observed. Gene expression revealed down-regulation of both SOCS-1 and SOCS-3 genes in resistant cells. STAT3 was phosphorylated in K562-R but not K562. Conclusions: Development of cells resistant to imatinib is feasible by overexposure of the drug to the cells. Activation of STAT3 protein leads to uncontrolled cell proliferation in imatinib resistant BCR-ABL due to DNA methylation of the SOCS-3 gene. Thus SOCS-3 provides a suitable candidate for mechanisms underlying the development of imatinib resistant in CML patients.