Ras Mitogen-activated Protein Kinase Signaling and Kinase Suppressor of Ras as Therapeutic Targets for Hepatocellular Carcinoma

( Hyuk Moon ),( Simon Weonsang Ro ) 대한간암학회 2021 대한간암학회지 Vol.21 No.1

Hepatocellular carcinoma (HCC) is a high incidence cancer and a major health concern worldwide. Among the many molecular signaling pathways that are dysregulated in HCC, the Ras mitogen-activated protein kinase (Ras/Raf/MAPK) signaling pathway has gained renewed attention from basic and clinical researchers. Mutations in Ras and Raf genes which are known to activate the Ras/Raf/MAPK signaling pathway have been infrequently detected in human HCC; however, the Ras/Raf/MAPK signaling pathway is activated in more than 50% of HCC cases, suggesting an alternative mechanism for the activation of the signaling pathway. Kinase suppressor of Ras acts as a molecular scaffold for facilitating the assembly of Ras/Raf/MAPK signaling pathway components and has been implicated in the regulation of this signaling pathway. In this review, we provide important insights into the cellular and molecular mechanisms involved in the activation of the Ras/Raf/MAPK signaling pathway and discuss potential therapeutic strategies for HCC. (J Liver Cancer 2021;21:1-11)

Sleeping Beauty transposon system harboring HRAS, c-Myc and shp53 induces sarcomatoid carcinomas in mouse skin

JUNG, SUNYOUNG,RO, SIMON WEONSANG,JUNG, GEUNYOUNG,JU, HYE-LIM,YU, EUN-SIL,SON, WOO-CHAN D.A. Spandidos 2013 Oncology Reports Vol.29 No.4

<P>The Sleeping Beauty transposon system is used as a tool for insertional mutagenesis and oncogenesis. However, little is known about the exact histological phenotype of the tumors induced. Thus, we used immunohistochemical markers to enable histological identification of the type of tumor induced by subcutaneous injection of the HRAS, c-Myc and shp53 oncogenes in female C57BL/6 mice. The tumor was removed when it reached 100 mm<SUP>3</SUP> in volume. Subsequently, we used 13 immunohistochemical markers to histologically identify the tumor type. The results suggested that the morphology of the tumor was similar to that of sarcomatoid carcinoma.</P>

모바일 쇼핑에서의 소비자 문화적 혁신성 척도개발

송니은(Nieun Song),유원상(Weonsang Yoo),김보영(Renee Boyoung Kim) 대한경영학회 2020 大韓經營學會誌 Vol.33 No.10

LC, Acute : PE-112 ; Combined effects of caspase inhibitor Nivocasan and lithospermate B on the inhibition of hepatic fibrosis in rats

( Do Young Kim ),( Sook In Chung ),( Weonsang Ro ),( Yong Han Paik ),( Young Nyun Park ),( Sang Hoon Ahn ),( Jung Gyu Park ),( Hee Dong Park ),( Kwan Sik Lee ),( Kwang Hyub Han ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.-

Background/Aim: To see combined anti-fibrosis effect when lithospermate B (LAB), an anti-oxidant, and Nivocasan, a caspase inhibitor, were simultaneously administered in comparison with either compound. Methods: Hepatic fibrosis was induced in SD rats by thioacetamide (TAA). In fibrosis-preventive experiment, rats were injected with TAA and fed on LAB and Nivocasan at the same time. Grouping was; Normal control (N), Normal+ LAB+ Nivocasan (NLN), TAA control (T), TAA+LAB (TL), TAA+ Nivocasan (TN), TAA+LAB+Nivocasan (TLN). In fibrosis- reversal experiment, rats were first treated with TAA, and then LAB and Nivocasan were fed. Grouping was; TAA control (F), TAA+LAB (FL), TAA+Nivocasan (FN), and TAA+LAB+ Nivocasan (FLN). Fibrotic area was evaluated quantitatively by computerized morphometry. Apoptosis was assessed using TUNEL assay, and immunohistochemical staining for malondialdehyde (MDA) and 4-hydroxy-2-nonenal (4HNE) was performed to assess oxidative stress. Real-time quantitative PCR was used to determine expression of fibrosis-related genes. Results: The degree of hepatic fibrosis was significantly reduced in group TLN (4.9%) compared to TL (6.3%) or TN (6.7%) (p < 0.001). These results were similarly observed in fibrosis- reversal experiment. Treatment with each compound significantly decreased fibrosis-related gene expression such as type I collagen α1 (col1α1), α-SMA, and TGF-β1 (p < 0.05). Co-treat- ment with LAB and Nivocasan further reduced col1α1 expression compared with either compound in both fibrosis- preventive and reversal experiment. TUNEL assay revealed that hepatocyte apoptosis significantly decreased in group TN and TLN compared to TL (p < 0.001). Similarly, apoptosis reduced significantly in group FN and FLN compared to FL (p < 0.001). Immunohistochemistry showed decrease of MDA and 4HNE, reflecting amelioration of oxidative stress in both fibrosis- preventive and reversal experiment, when LAB or LAB+ Nivocasan was administered compared with Nivocasan alone (p < 0.05). Conclusion: Simultaneous administration of LAB and Nivocasan to suppress oxidative stress and apoptosis resulted in an enhanced effect on inhibition of hepatic fibrosis in rats.

Basic, HCC basic : PE-100 ; Development of various double transgenic Liver cancer models by hydrodynamic transfection

( Hye Lim Ju ),( Kwang Hyub Han ),( Weonsang S. Ro ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.-

Background/Aims: Liver cancer is a major health concern worldwide. Liver cancer is known to develop through complicated interactions of oncogenes and tumor suppressor genes. However, its pathogenesis and underlying mechanisms are incompletely understood. In this study, various double transgenic liver cancer mouse models have been developed using a hydrodynamic injection method and the Sleeping Beauty transposon system. The strategy allows a double transgenic mouse to be developed efficiently reducing time and resources significantly. Using the double transgenic liver cancer models, we could detect tumors noninvasively in vivo and reveal genetic influences on the development of liver cancer. Methods: Transposon vectors each encoding an oncogene or down-regulating a tumor suppressor gene (SmoM2, HrasG12V, Ctnnb1S33Y and shp53) were constructed. To induce liver cancer, total 50 μg of the three plasmids - encoding the Sleeping Beauty transposase and two transposons - were diluted in 2.5 ml of 0.9% saline and then injected into the lateral tail veins of 6- to 8-week-old C57BL/6 mice. Tumors were observed via bioluminescence imaging. Results: Liver cancer was induced by the stable expression of the transposed oncogenes into the mouse chromosome. Tumor growths were observed via repeated bioluminescence imaging of mice over time. Whereas control mice that stably expressed only luciferase showed no significant changes in the luciferase signal intensities over time, the mice expressing oncogenes in the liver exhibited continuous tumor growth over months, based on the luciferase signal intensities. Conclusion: Established double transgenic mouse models could be an efficient alternative to traditional transgenic mouse models for liver cancer with significantly reduced time and cost.

Pro-tumorigenic roles of TGF-β signaling during the early stages of liver tumorigenesis through upregulation of Snail

( Hyuk Moon ),( Kwang-hyub Han ),( Simon Weonsang Ro ) 생화학분자생물학회(구 한국생화학분자생물학회) 2017 BMB Reports Vol.50 No.12

Many studies have focused on the tumor suppressive role of TGF-β signaling during the early stages of tumorigenesis by activating the target genes involved in cytostasis and apoptosis. We investigated the effects of TGF-β inhibition on early tumorigenesis in the liver, by employing diverse inhibitory methods. Strikingly, TGF-β inhibition consistently suppressed hepatic tumorigenesis that was induced either by activated RAS plus p53 downregulation or by the co-activation of RAS and TAZ signaling; this demonstrates the requirements for canonical TGF-β signaling in tumorigenesis. Moreover, we found that Snail is the target gene of the TGF-β signaling pathway that promotes hepatic carcinogenesis. The knockdown of Snail suppressed the early tumorigenesis in the liver, as did the TGF-β inhibition, while the ectopic expression of Snail restored tumorigenesis that was suppressed by the TGF-β inhibition. Our findings establish the oncogenic TGF-β-Smad- Snail signaling axis during the early tumorigenesis in the liver. [BMB Reports: Perspective 2017; 50(12): 599-600]

HCV : PE-100 ; Development of various double transgenic liver cancer models by hydrodynamic transfection

( Hye Lim Ju ),( Kwang Hyub Han ),( Weonsang S. Ro ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1

Background/Aims: Liver cancer is a major health concern worldwide. Liver cancer is known to develop through complicated interactions of oncogenes and tumor suppressor genes. However, its pathogenesis and underlying mechanisms are incompletely understood. In this study, various double transgenic liver cancer mouse models have been developed using a hydrodynamic injection method and the Sleeping Beauty transposon system. The strategy allows a double transgenic mouse to be developed efficiently reducing time and resources significantly. Using the double transgenic liver cancer models, we could detect tumors noninvasively in vivo and reveal genetic influences on the development of liver cancer. Methods: Transposon vectors each encoding an oncogene or down-regulating a tumor suppressor gene (SmoM2, HrasG12V, Ctnnb1S33Y and shp53) were constructed. To induce liver cancer, total 50 μg of the three plasmids - encoding the Sleeping Beauty transposase and two transposons - were diluted in 2.5 ml of 0.9% saline and then injected into the lateral tail veins of 6- to8-week-old C57BL/6 mice. Tumors were observed via bioluminescence imaging. Results: Liver cancer was induced by the stable expression of the transposed oncogenes into the mouse chromosome. Tumor growths were observed via repeated bioluminescence imaging of mice over time. Whereas control mice that stably expressed only luciferase showed no significant changes in the luciferase signal intensities over time, the mice expressing oncogenes in the liver exhibited continuous tumor growth over months, based on the luciferase signal intensities. Conclusion: Established double transgenic mouse models could be an efficient alternative to traditional transgenic mouse models for liver cancer with significantly reduced time and cost.

간암 동물모델의 현황 및 전임상 연구에서의 활용

주혜림 ( Hye-lim Ju ),노원상 ( Simon Weonsang Ro ) 대한간암학회 2017 대한간암학회지 Vol.17 No.1

Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide. HCC develops in various causes - Viral hepatitis infection, toxins, or other liver conditions - by activation of oncogenes and/or inactivation of tumor suppressors. Understanding of signal pathways and protein-protein interactions critical in tumor development may lead to novel treatment strategy. To evaluate the progression of HCC and effects of potential therapies, various animal models have been established. Experimental models of HCC provide valuable tools to investigate the risk factors, new treatment modalities and biologic characteristics. Subcutaneous xenograft models have been widely used in the past. However, with the advancement of in vivo imaging technology, investigators are more concerned with the orthotopic models nowadays. Genetically engineered mouse models have greatly facilitated studies of gene function in HCC development. Lately, a novel approach for stable gene expression in mouse hepatocytes by hydrodynamic injection has been developed. Each model has its own advantages and disadvantages. Therefore, selecting the optimal models based on study objectives is necessary. In this review, we highlight both the frequently used mouse models and some emerging ones with emphasis on their merits or defects, and give advices for investigators to choose a ``best-fit`` animal model in HCC research. (J Liver Cancer 2017;17:1-14)

Synergic chemoprevention with dietary carbohydrate restriction and supplementation of AMPK-activating phytochemicals: the role of SIRT1

Lee, Jong Doo,Choi, Min-Ah,Ro, Simon Weonsang,Yang, Woo Ick,Cho, Arthur E.H.,Ju, Hye-Lim,Baek, Sinhwa,Chung, Sook In,Kang, Won Jun,Yun, Mijin,Park, Jeon Han Lippincott WilliamsWilkins 2016 EUROPEAN JOURNAL OF CANCER PREVENTION Vol.25 No.1

<P>Calorie restriction or a low-carbohydrate diet (LCD) can increase life span in normal cells while inhibiting carcinogenesis. Various phytochemicals also have calorie restriction-mimetic anticancer properties. We investigated whether an isocaloric carbohydrate-restriction diet and AMP-activated protein kinase (AMPK)-activating phytochemicals induce synergic tumor suppression. We used a mixture of AMPK-activating phytochemical extracts including curcumin, quercetin, catechins, and resveratrol. Survival analysis was carried out in a B16F10 melanoma model fed a control diet (62.14% kcal carbohydrate, 24.65% kcal protein and 13.2% kcal fat), a control diet with multiple phytochemicals (MP), LCD (16.5, 55.2, and 28.3% kcal, respectively), LCD with multiple phytochemicals (LCDmp), a moderate-carbohydrate diet (MCD, 31.9, 62.4, and 5.7% kcal, respectively), or MCD with phytochemicals (MCDmp). Compared with the control group, MP, LCD, or MCD intervention did not produce survival benefit, but LCDmp (22.80±1.58 vs. 28.00±1.64 days, <I>P</I>=0.040) and MCDmp (23.80±1.08 vs. 30.13±2.29 days, <I>P</I>=0.008) increased the median survival time significantly. Suppression of the IGF-1R/PI3K/Akt/mTOR signaling, activation of the AMPK/SIRT1/LKB1pathway, and NF-κB suppression were the critical tumor-suppression mechanisms. In addition, SIRT1 suppressed proliferation of the B16F10 and A375SM cells under a low-glucose condition. Alterations in histone methylation within <I>Pten</I> and <I>FoxO3a</I> were observed after the MCDmp intervention. In the transgenic liver cancer model developed by hydrodynamic transfection of the <I>HrasG12V</I> and <I>shp53</I>, MCDmp and LCDmp interventions induced significant cancer-prevention effects. Microarray analysis showed that <I>PPARα</I> increased with decreased <I>IL-6</I> and <I>NF-κB</I> within the hepatocytes after an MCDmp intervention. In conclusion, an isocaloric carbohydrate-restriction diet and natural AMPK-activating agents induce synergistic anticancer effects. SIRT1 acts as a tumor suppressor under a low-glucose condition.</P>

Basic, Research : Rapid Induction of Liver Cancer using Hydrodynamic Transfection of Oncogene-encoding Plasmids

( Hye Lim Ju ),( Sang Hoon Ahn ),( Do Young Kim ),( Sinhwa Baek ),( Sook In Chung ),( Kwang Hyub Han ),( Simon Weonsang Ro ) 대한간학회 2013 춘·추계 학술대회 (KASL) Vol.2013 No.1

Background : Liver cancer is a complex multistep process requiring genetic alterations in multiple proto-oncogenes and tumor suppressor genes. Although hundreds of genes are known to play roles in hepatocarcinogenesis, oncogenic collaboration among these genes is still largely unknown. We developed various non-germline transgenic mouse models using hydrodynamics- based transfection and the Sleeping Beauty transposon system. Here, we report a method by which oncogenic collaboration of various cancer-related genes in the liver can be easily investigated in vivo by bioluminescence imaging (BLI) of tumors. Methods: Transposon vectors each encoding an oncogene or down-regulating a tumor suppressor gene (HrasG12V, SmoM2, and shp53) were constructed. To induce liver cancer, total 50 μg of the three plasmids-encoding the Sleeping Beauty transposase and two transposons-were diluted in 2.5 mL of 0.9% saline and then injected into the lateral tail veins of male 5- to 6-week-old C57BL/6 mice. Tumors were observed via BLI. Results: Very strong BLI signals were observed at 4 weeks post-hydrodynamic injection (PHI) in mice co-expressing HrasG12V and shp53, while only background signals were detected in other double or single transgenic groups until 30 weeks PHI. Consistent with the BLI data, tumors were observed in the HrasG12V plus shp53 group at 4 weeks PHI, while other transgenic groups failed to exhibit a hyperplastic nodule at 30 weeks PHI. Conclusion: The methodology described here is expected to accelerate and facilitate in vivo studies of the hepatocarcino genic potential of cancer-related genes by means of oncogenic cooperation.