• Asian Pacific Journal of Cancer Prevention
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• 제16권13호
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• pp.5343-5348
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• 2015

Promoter hypermethylation of the runt-related transcription factor 3 (RUNX3) gene is associated with increased risk of hepatocellular carcinoma (HCC). Oxidative stress plays a vital role in both carcinogenesis and progression of HCC. However, whether oxidative stress and RUNX3 hypermethylation in HCC have a cause-and-effect relationship is not known. In this study, plasma protein carbonyl and total antioxidant capacity (TAC) in patients with hepatitis B virus (HBV)-associated HCC (n=60) and age-matched healthy subjects (n=80) was determined. RUNX3 methylation in peripheral blood mononuclear cells (PBMC) of subjects was measured by methylation-specific PCR. Effect of reactive oxygen species (ROS) on induction of RUNX3 hypermethylation in HCC cells was investigated. Plasma protein carbonyl content was significantly higher, whereas plasma TAC was significantly lower, in HCC patients than healthy controls. Based on logistic regression, increased plasma protein carbonyl and decreased plasma TAC were independently associated with increased risk for HCC. PBMC RUNX3 methylation in the patient group was significantly greater than in the healthy group. RUNX3 methylation in hydrogen peroxide ($H_2O_2$)-treated HepG2 cells was significantly higher than in untreated control cells. In conclusion, increase in oxidative stress in Thai patients with HBV-associated HCC was demonstrated. This oxidative increment was independently associated with an increased risk for HCC development. RUNX3 in PBMC was found to be hypermethylated in the HCC patients. In vitro, RUNX3 hypermethylation was experimentally induced by $H_2O_2$. Our findings suggest that oxidative stress is a cause of RUNX3 promoter hypermethylation in HCC cells.

• 대한한방내과학회지
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• 제20권1호
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• pp.122-132
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• 1999

Purpose : Hepatitis B virus DNA transfected cell line(HepG2.2.15) was cultured to evaluate the effect of herbs on the expression of HBeAg and the replication of HBV. HepG2.2.15 produces HBV particles as well as viral proteins into cell culture media. Methods : Extracts of herbs were adminitered to the cells on the proper concentration. Culture media was collected 48 hours after the herbal administration and HBeAg level in the media was examined by ELISA method. To confirm that the anti-viral effect was not due to direct cytotocixity of the extracts, normal cell proliferation was shown by cell counting. And as of the interference in protein synthesis of HepG2.2.15 by herb-extracts, we used the result of study that we performed before by ${\alpha}FP$ assay using EIA method. Results& Conclusion : Herb medicines like 地楡(Sanguisorbae Radix) and 覆盆子(Rubi Frusctus) showed significant inhibitory effect on HBeAg expression at p<0.01 and 五味子(Acanthopanacis Cortex) at p<0.05. Whereas, though some herbs such as ?草根(Rubiae Radix), 山査(Crataegii Fructus), 白芍藥(Paeoniae Radix Alba), and 大黃(Rhei Radix et Rhizoma) showed the tendecy to suppress HBeAg. most of them were not significant statistically. From the above, we could conclude that those herb medicines can be applied to patients effectively and further studies on effective fraction of some herbs are thought to be needed.

• 대한한방내과학회지
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• 제37권3호
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• pp.529-538
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• 2016

Objectives: The purpose of this case study is to report the efficacy and safety of treatment with Korean medicine of a patient with HBeAg-positive chronic viral hepatitis B.Methods: The patient took Korean medicine (mainly Injinchunggan-tang-gamibang) from July 20th, 2010, to March 14th, 2016, without any antiviral or interferon therapy. Changes to laboratory records, abdomen ultrasonography, and clinical symptoms were reviewed.Results: The laboratory records showed that AST, ALT, and HBV DNA had decreased to normal ranges, and HBeAg showed seroconversion. Clinical symptoms also improved after taking Korean medicine.Conclusion: The results suggest that treatment with Korean medicine and without antiviral or interferon therapy could be effective for HBeAg-positive chronic hepatitis B.

• 한국약용작물학회지
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• 제17권2호
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• pp.133-138
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• 2009

Nelumbo nucifera (lotus) is known to be a useful medicinal plant and leaf extract contains several flavonoids and alkaloids. To analyze the effect of Nelumbo nucifera leaves extract (NNL) on the HBsAg production, we treated NNL on HepG2.2.15 cells which contain the hepatitis B viral genome and secrete surface antigen into media. NNL suppressed the production of hepatitis B surface antigen as a dose-dependent manner. To analyze the effect of NNL on HBV DNA replication, PCR analysis was performed. NNL was not affected the HBV DNA replication and HBsAg mRNA expression. To understand the effect of NNL on the production of HBsAg, we carried out the analysis of lipid-metabolizing gene expression using one-step RT-PCR. NNL reduced the gene expression of FASN and SREBP2 and increased the expression of LDLR. Triglyceride content of HepG2.2.15 cells was not decreased by treatment of NNL. This result suggests a possibility that NNL may have an effect for the inhibition of hepatitis B surface antigen by modulation of lipid and cholesterol metabolism.

• IMMUNE NETWORK
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• 제9권1호
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• pp.20-26
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• 2009

We previously reported that $IFN-{\gamma}$ producing T cell responses induced by the combined therapy of DNA vaccine and lamivudine for one year are important for the induction of sustained virological response (SVR). However, $IFN-{\gamma}$ production is not sufficient to predict sustained viremia control in chronic hepatitis B (CHB) carriers treated. Methods: Twelve CHB carriers were intramuscularly immunized 12 times at a 4-week interval with 8mg of HBV DNA vaccine during the standard lamivudine treatment (100mg/daily/1 year). The level of cytokines during and after the combined therapy in plasma of all 12 CHB carriers treated was determined by each ELISA kit. Six out of 12 CHB carriers revisited the clinic, and their HBV DNA levels were examined. Results: The combined therapy increased plasma IL-12 and IL-12/p40 ratio during the treatment (baseline vs. peak level: $41.8{\pm}8.3$ vs. $163.1{\pm}29.2\;pg/ml$; p<0.01 and $0.96{\pm}0.25$ vs. $3.58{\pm}0.86$; p<0.01, respectively), and the peak level of plasma IL-12 and IL-12/p40 ratio was evoked at 6 to 10 months during the combined therapy. In particular, CHB carriers with SVR had two and three-fold higher level of the peak plasma IL-12 and plasma IL-12/p40 ratio than non-virological responders (NVRs), respectively ($218.0{\pm}41.4$ vs. $108.1{\pm}28.6\;pg/ml$; p=0.09 and $5.35{\pm}1.38$ vs. $1.80{\pm}0.29$; p<0.05, respectively), while p40 level was consistent during the combined therapy. In addition, there was no significant temporal correlation between the peak IL-12/p40 ratio and the elevation of serum alanine amino-transferase (ALT) in this study, contrast to $IFN-{\alpha}$ therapy which induced peak IL-12 level following ALT flares. Conclusion: Our results indicate that the combined therapy induces the increase of plasma IL-12 and IL-12/p40 ratio, which are associated with long-term SVR in CHB carriers.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제7권2호
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• pp.197-207
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• 2004

목적: 소아 만성 B형 간염 환아들에게 라미부딘으로 치료 시작 후 그 치료 효과를 평가하고 소아에서 라미부딘 장기 치료의 지속성과 안정성에 대해 검증하고자 하였다. 대상 및 방법: 1999년 3월부터 경북대학교병원 소아과에서 만성 B형 간염으로 라미부딘 치료를 시작한 후 2004년 9월 현재까지 최소 6개월 이상 치료한 48명(남 31, 여 17명, 1~18세, 평균 8세)을 대상으로 29개월(8~66개월) 추적 관찰하면서 연구를 시행하였다. 치료에 대한 효과는 치료 시작 후 혈청 ALT 치의 정상화와 HBV DNA의 음전 및 HBeAg/anti-HBe로의 혈청전환을 모두 만족할 때 치료반응이 있다고 정의하였다. Kaplan-Meier법을 이용한 누적 HBeAg 혈청전환율을 치료 시작 0.5, 1, 1.5, 2, 2.5, 3년에서 구하였다. 결과: 라미부딘으로 치료 시작 후 0.5년이 경과한 48명 중 치료 반응은 29명(60%)에서 보였고 9명(19%)에서는 HBsAg의 소실도 일어났다. 치료 시작 1년째 ALT치가 정상화된 환아는 94%, HBV DNA치가 음전된 환아는 94%, HBeAg 혈청전환까지 되어 치료 반응이 있던 환아는 34%였다. Kaplan-Meier법에 의한 누적 HBeAg 혈청전환율은 0.5, 1, 1.5, 2, 2.5, 3년째에 각각 13, 34, 50, 68, 79, 90%로 계산되었다. 특히 7세 미만의 소아 22명 중에서는 HBsAg의 소실이 8명(36%)에서 일어나 7세 이상에서 치료를 시작하는 것에 비하여 탁월한 성적을 보여주었다(p=0.002). 결론: 한국의 소아 만성 B형 간염 환아에서 라미부딘의 장기 치료는 HBeAg 혈청전환을 가속화 시키며 3년 동안 추적 관찰 결과 장기적으로 치료 반응이 지속되었다. 소아 만성 간염의 경과 관찰 중 면역제거기에 들어서면 라미부딘으로 장기간 적극적인 치료를 시도하는 것이 좋을 것으로 생각한다.

• Molecules and Cells
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• 제40권6호
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• pp.418-425
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• 2017

Interferon-${\gamma}$-inducible protein 10 (IP-10), also known as chemokine C-X-C motif ligand (CXCL) 10, is closely associated with antiviral immunity and the progression of chronic hepatitis B (CHB). However, the value of baseline serological and histological IP-10 expression levels in predicting the efficacy of the antiviral response to nucleoside/nucleotide analogues (NAs) is still unknown. In our research, intrahepatic and peripheral IP-10 expression levels were systemically examined before and after treatment with entecavir (ETV). Baseline serological and histological IP-10 expression levels were significantly increased in patients with CHB, particularly in patients with higher degrees of liver inflammation and liver fibrosis. Moreover, higher baseline intrahepatic IP-10 levels indicated better prognoses in patients with CHB after entecavir therapy. The baseline IP-10 level was also positively associated with several clinical parameters, including baseline levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), hepatitis B virus (HBV) DNA, and hepatitis B surface antigen (HBsAg), and with the decrease in HBsAg levels after treatment. In addition, monocyte-derived IP-10 was expressed at higher levels in patients with CHB than in patients with liver cirrhosis (LC) and healthy controls (HC). According to the results of our in vitro experiments, IP-10 directly promoted hepatocyte apoptosis. Based on these findings, baseline serological and histological IP-10 levels might predict CHB severity and the decrease in HBsAg levels after entecavir therapy.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제6권2호
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• pp.152-160
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• 2003

목 적: 소아의 만성 B형 간염 치료에 interferon-${\alpha}$의 일정한 치료 효과가 보고되고 있다. 저자들은 interferon-${\alpha}$의 용량 차이에 따른 치료 효과 및 초치료와 재치료에 따른 치료 효과에 차이가 있는지 비교해 보았다. 방 법: 1990년 3월부터 1999년 8월까지 경북대학교병원 소아과에 내원하였던 환아(2~14세) 중 6개월 이상 HBsAg, HBeAg 및 HBV DNA가 양성이고, 혈청 ALT치가 상승되어 있는 51명의 환아를 대상으로 27명에게 interferon-${\alpha}$ $3MU/m^2$ ($2.66{\pm}0.66\;MU/m^2$)를 투여하였고 24명에게는 $6\;MU/m^2$ ($4.45{\pm}0.94\;MU/m^2$)을 주 3회씩 6개월(6~12개월)간 피하 혹은 근육 주사하였다. interferon-${\alpha}$ 초치료 평균용량은 $3.50{\pm}1.20\;MU/m^2$이었고 평균 치료 기간은 7개월(6~12개월)이었다. 초치료에 반응이 없었던 환아 중 12명을 대상으로 다시 interferon-${\alpha}$ 재치료를 시행하였다. 재치료 평균 용량은 $3.62{\pm}1.51\;MU/m^2$이었고 평균 치료 기간은 7개월(6~12개월)이었다. 용량 차이를 보인 두 군 사이에 성별, 연령, 치료기간, 치료전 ALT치와 HBV DNA 등에서는 통계학적으로 유의한 차이가 없었으며 초치료, 재치료 두 군간에도 유의한 차이가 없었다. 결 과: 치료 시작 1년 후 시점에서 interferon $3\;MU/m^2$로 치료한 27명 중 11명(41%)에서 ALT의 정상화를 보였고 9명(33%)에서 HBeAg이 anti-HBe로 혈청전환이 되었다. 한편 $6\;MU/m^2$ 치료군 24명 중 에서는 12명(50%)에서 ALT의 정상화를 보였고 7명(29%)에서 혈청전환이 되었는데 두 군 사이의 치료성적에는 통계학적으로 유의한 차이가 없었다. Interferon $3\;MU/m^2$ 치료군에서 발생한 부작용으로는 발열 14례(52%), 백혈구 감소증 10례(37%)였으며 모든 경우에서 특별한 조치 없이 회복되었다. 한편 interferon $6\;MU/m^2$ 치료군에서는 발열 16례(67%), 백혈구 감소증 8례(33%), 혈소판 감소증 1례(4%), 갑상선 기능 저하증 2례(8%)가 있었다. 치료와 관련된 부작용도 두 군 간에 통계학적으로 유의한 차이가 없었다. Interferon-${\alpha}$ 초치료군 51명 중 23명(45%)에서 ALT의 정상화를 보였고 16명(31%)에서 혈청전환이 있었으며 재치료군은 12명 중 3명(25%)에서 ALT의 정상화 및 혈청전환이 있었다. 결 론: Interferon-${\alpha}$ $3\;MU/m^2$ 치료군과 $6\;MU/m^2$치료군을 비교했을 때 ALT의 정상화 및 혈청전환에서 의미 있는 치료 효과의 차이를 찾을 수 없었다. interferon-${\alpha}$ 재치료는 초치료만큼의 효과가 있음을 보여주었다.

• The Korean Journal of Physiology and Pharmacology
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• 제8권4호
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• pp.213-218
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• 2004

Primary fish-odor syndrome (FOS) is a genetic disorder caused by defective flavin-containing mono-oxygenase 3 gene (FMO3) with deficient N-oxidation of trimethylamine (TMA), causing trimethylaminuria (TMAU). By contrast, secondary FOS can be acquired by decreased FMO activities in patients with chronic liver diseases, but the underlying mechanisms are unknown. In the present study, we examined plasma NOx concentrations and viral DNA contents as well as in vivo FMO activities and their correlations in chronic viral hepatitis (CVH) patients. Plasma concentration of NOx was significantly increased by 2.1 fold $(56.2{\pm}26.5\;vs.\;26.6{\pm}5.4\;{\mu}M,\;p<0.01)$, and it was positively correlated with plasma hepatitis B virus (HBV) DNA contents $(r^2=0.2838,\;p=0.0107)$. Furthermore, the elevated plasma NOx values were inversely and significantly correlated with in vivo FMO activities detected by ranitidine-challenged test $(8.3%\;vs.\;20.0%,\;r^2=0.2109,\;p=\0.0315)$. TMA N-oxidation activities determined in CVH patients without challenge test were also significantly low (73.6% vs. 95.7%, p< 0.05). In conclusion, these results suggested that secondary FOS could be acquired by the endogenously elevated NO in patients with CVH.

• BMB Reports
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• 제52권8호
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• pp.520-525
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• 2019

Dihydroartemisinin (DHA) has been reported to possess anti-cancer activity against many cancers. However, the pharmacologic effect of DHA on HBV-positive hepatocellular carcinoma (HCC) remains unknown. Thus, the objective of the present study was to determine whether DHA could inhibit the proliferation of HepG2.2.15 cells and uncover the underlying mechanisms involved in the effect of DHA on HepG2.2.15 cells. We found that DHA effectively inhibited HepG2.2.15 HCC cell proliferation both in vivo and in vitro. DHA also reduced the migration and tumorigenicity capacity of HepG2.2.15 cells. Regarding the underlying mechanisms, results showed that DHA induced cellular senescence by up-regulating expression levels of proteins such as p-ATM, p-ATR, ${\gamma}-H_2AX$, P53, and P21 involved in DNA damage response. DHA also induced autophagy (green LC3 puncta gathered together and LC3II/LC3I ratio increased through AKT-mTOR pathway suppression). Results also revealed that DHA-induced autophagy was not linked to senescence or cell death. TPP1 (telomere shelterin) overexpression could not rescue DHA-induced anticancer activity (cell proliferation). Moreover, DHA down-regulated TPP1 expression. Gene knockdown of TPP1 caused similar phenotypes and mechanisms as DHA induced phenotypes and mechanisms in HepG2.2.15 cells. These results demonstrate that DHA might inhibit HepG2.2.15 cells proliferation through inducing cellular senescence and autophagy.