Basic, Research : Expression Levels of Cancer-related microRNAs in Human Hepatocellular Carcinoma (HCC)

( Sinhwa Baek ),( Jun Yong Park ),( Do Young Kim ),( Sang Hoon Ahn ),( Kwang Hyub Han ),( Weon Sang S Ro ) 대한간학회 2013 춘·추계 학술대회 (KASL) Vol.2013 No.1

Background: Hepatocellular carcinoma (HCC) is the third cause of cancer-related death worldwide. Curative options for HCC are only available for patients with tumor of an early stage, thus a precise method to detect HCC in its early stage is needed. MicroRNAs (miRNAs) are non-coding small (~25nt) RNAs that play important roles in gene regulatory networks by repressing the activity of specific target mRNAs. Aberrant expression of several miRNAs was found to be involved in human hepatocarcinogenesis. In this study we compared the expression levels of miRNAs between surgically resected HCC tissues and surrounding non-tumor tissues of 20 patients with HCC of early stages. Aims: The aim of this study is to identify miRNAs that can be used as biomarkers for an early stage HCC. Methods: miRNAs were isolated from surgically resected HCC tissues and surrounding non-tumor tissues of 20 HCC patients and used to synthesize cDNAs. We designed primers for 50 kinds of miRNAs, selection of which was based on published microarray studies. We performed SYBR Green quantitative RT-PCR (qRT-PCR) assays to compare expression levels of miRNAs between HCC and surrounding non-HCC tissues. Results: Six miRNAs were found highly up-regulated in early stage HCC. MiR-17-5p, 24, 25, 107, 221, and 222 were significantly up-regulated in 70%, 65%, 75%, 75%, 90%, and 85% of HCC samples, respectively. All of the HCC samples, at least one of miR-221 and miR-25 was up-regulated, as compared with adjacent non-tumor tissues. For mechanism study, we transfected the miRNAs expressing plasmids in HCC cell lines. MiR-221 enhanced cell proliferation in Huh7 cells. And miR- 221, 21-5p, 25, 222, and 17-5p are increased cell migration in Hep3B cells. Conclusions: We identified miRNAs which are significantly up-regulated in early stage HCC samples. All patients had an elevated expression level of either miR-221 or miR-25. We suggest that the combination of the two microRNAs might be an effective biomarker for detecting early HCC.

Inhibitory Effects of Fermented Black Radish (Raphanus sativus L. var niger) on Antioxidant and Lipid Accumulation

Seong-Eun Kim,Sinhwa Baek,Hak Sung Lee,Hyun-Kyu Kim 한국식품영양과학회 2018 한국식품영양과학회 학술대회발표집 Vol.2018 No.10

유산균 발효 흑무의 항산화 및 지방구 형성 억제 효과

김성은(Seong-Eun Kim),백신화(Sinhwa Baek),이학성(Hak Sung Lee),김현규(Hyun-Kyu Kim) 한국식품영양과학회 2018 한국식품영양과학회지 Vol.47 No.7

본 연구는 Lactobacillus plantarum을 이용하여 발효시킨 흑무 유산균 발효물의 간 손상 보호 효과를 알아보기 위해 기초적인 항산화 효능 및 간세포 지방구 형성 억제 활성 실험을 하였다. 흑무 유산균 발효물은 단순 추출물 및 발효 전 단계의 영양원과 비교하여 유의적으로 증진된 항산화 효능을 보였으며, 총 폴리페놀 함량 및 총 플라보노이드 함량 역시 발효를 통해 증가한 것을 확인하였다. 또한, 간 손상 유발 시 간세포에 형성되는 지방구의 형성 억제를 통한 간세포 보호 효능도 단순 추출물 및 발효 전 단계 영양원과 대비하여 유의적으로 증가한 것을 확인하였다. 이상의 결과를 통해 흑무 유산균 발효물은 종전의 흑무 추출물과 대비하여 손상에 의한 간세포의 지방구 형성 억제 효능이 증진되었으며 이를 토대로 유산균 발효 흑무는 간 건강 개선을 위한 건강기능식품 소재로써 활용될 수 있는 가능성을 확인하였다. Black radish (Raphanus sativus var. niger) has been reported to have anti-oxidant, anti-inflammatory and detoxification effects. In this study, we investigated the antioxidant and lipid accumulation inhibitory effects of fermented black radish (FBR) with Lactobacillus plantarum compared with non-fermented black radish (BR) and black radish extracts obtained using water, and 30%, 50%, 70% ethyl alcohol. The lipid accumulation inhibitory effect was determined by Oil-Red O staining activity after oleic acid treatment of HepG2 cells. Treatment with FBR at doses of 100 and 500 μg/mL inhibited lipid accumulation by 103% and 108% and BR at the same dose inhibited lipid accumulation by 72% and 61%, respectively. Extracts inhibited lipid accumulation ranging by 17% to 93% when applied in the same doses. FBR led to significant inhibition of lipid accumulation compared with BR and extracts. In addition, FBR demonstrated antioxidant effect such as scavenging activity of (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 2,2-diphenyl-1-picrylhydrazyl radical and super oxide dismutase (SOD)-like activity. Also the FBR was measured total polyphenol contents (TPC) and total flavonoid contents (TFC). Moreover, FBR showed significant greater antioxidant activity than BR and extracts. The TPC of FBR increased more than BR and the TFC of FBR was higher than that of other extracts. These results suggest that FBR might be a potential health functional food ingredient for enhancing liver function.

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.

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>