• Clinical and Experimental Reproductive Medicine
• /
• 제38권4호
• /
• pp.193-202
• /
• 2011

Objective: We found previously that $interferon$ $regulatory$ factor ($Irf$)-1 is a germinal vesicle (GV)-selective gene that highly expressed in GV as compared to metaphase II oocytes. To our knowledge, the function of $Irf-1$ in oocytes has yet to be examined. The present study was conducted to determine the relationship between retinoic acid (RA) and RA-mediated expression of $Irf-1$ and the mouse oocyte maturation. Methods: Immature cumulus-oocyte-complexes (COCs) were collected from 17-day-old female mice and cultured $in$ $vitro$ for 16 hours in the presence of varying concentrations of RA (0-10 ${\mu}M$). Rate of oocyte maturation and activation was measured. Gene expression was measured by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and cytokine secretion in the medium was measured by Bio-Plex analysis. Apoptosis was analyzed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Results: The rates of oocyte maturation to metaphase II and oocyte activation increased significantly with RA treatment (10 nM-1 ${\mu}M$). With 100 nM RA treatment, lowest level of $Irf-1$ mRNA and cumulus cell's apoptosis was found. Among 23 cytokines measured by Bio-Plex system, the substantial changes in secretion of tumor necrosis factor-${\alpha}$, macrophage inflammatory protein-$1{\beta}$, eotaxin and interleukin-12 (p40) from COCs in response to RA were detected. Conclusion: We concluded that the maturation of oocytes and $Irf-1$ expression are negatively correlated, and RA enhances the developmental competence of mouse immature oocytes $in$ $vitro$ by suppressing apoptosis of cumulus cells. Using a mouse model, results of the present study provide insights into improved culture conditions for $in$ $vitro$ oocyte maturation and relevant cytokine production and secretion in assisted reproductive technology.

• Tuberculosis and Respiratory Diseases
• /
• 제75권2호
• /
• pp.59-66
• /
• 2013

Background: This study was conducted in order to elucidate the effects of docetaxel on the growth of peroxiredoxin 1 (Prx1) knockdown A549 xenograft tumors and further tested the role of Prx1 as a predictor for how a patient would respond to docetaxel treatment. Methods: Effects of docetaxel on the growth of scrambled- and shPrx1-infected A549 xenograft tumors in nude mice were measured. Moreover, immunohistochemical expression of Prx1 was evaluated in paraffin-embedded tissues from 24 non-small cell lung cancer patients who had received docetaxel-cisplatin regimens as a first-line treatment. Results: Docetaxel treatment in Prx1 knockdown xenograft tumor resulted in reduced tumors growth compared with other groups. Prx1 knockdown increased the production of cleaved caspases-8 and -9 in the control itself compared to scramble tumors. Moreover, docetaxel treatment in Prx1 knockdown tissue led to an increased protein band. Phosphorylated Akt was found in Prx1 scramble tissues. Phosphorylated FOXO1 was detected in the docetaxel treatment group. On the other hand, Prx1 knockdown completely suppressed the Akt-FOXO1 axis. The median progression-free survival (PFS) of patients with low Prx1 expression was 7 months (95% confidence interval [CI], 6.0-7.7), whereas the median progression-free survival of patients with high Prx1 expression was 4 months (95% CI, 4.0-5.0). However, high Prx1 expression was not associated with decreased PFS (p=0.114). Conclusion: Our findings suggest that elevated Prx1 provides resistance to docetaxel treatment through suppression of FOXO1-induced apoptosis in A549 xenograft tumors, but may not be related with the predictive significance for response to docetaxel treatment.

• Journal of Gastric Cancer
• /
• 제4권3호
• /
• pp.196-203
• /
• 2004

The Information Committee of the Korean Gastric Cancer Association sent questionnaires to 31 laparoscopic gastric surgeons about their personal experiences with laparoscopic gastric surgery from 2001 to 2003. Twenty-four surgeons responded to the questionnaires (response rate: $77.4\%$).The number of laparoscopic gastric surgeries from 2001 to 2003 was 1,130 and increased from 209 in 2001 to 593 in 2003. The number of operations for a gastric adenocarcinoma also increased from 87 cases in 2001 to 403 cases in 2003. Laparoscopic radical procedures, such as a laparoscopyassisted distal gastrectomy or total gastrectomy (LADG or LATG), have increased rapidly during this period. (55 cases in 2001, 150 cases in 2002, and 364 cases in 2003). Laparoscopic function-preserving gastrectomies were not performed until 2003, during which year one pylorus- preserving gastrectomy and six proximal gastrectomies were performed laparoscopically. A wedge resection for a gastric submucosal tumor was performed in 71 cases in 2001, 82 in 2002, and 103 in 2003. Hand-assisted laparoscopic surgery (HALS) was performed in 39 cases in 2001, 55 in 2002, and 49 in 2003. As for personal indications for a LADG, 14 surgeons performed a LADG only for a T1 lesion, and 5 surgeons extended their indications to T2N0 lesions. In the near future, laparoscopic procedures for gastric cancer will be widely adopted in Korea if the medical-insurance obstacle is overcome and the long-term survival results are verified.

• Journal of Acupuncture Research
• /
• 제31권4호
• /
• pp.81-97
• /
• 2014

Objectives : The purpose of this study is analyzing the current research methodology of pharmacopucture for the treatment of animal cancer models. Methods : Four electronic databases were searched for animal studies published from January 2000 to September 2014 onward using these search terms "cancer, anticancer, pharmacopuncture, beevenom". Selected articles were described about animal cancer models. The methods used to induce cancer and the outcome measures used to assess the effects of pharmacopuncture on animal cancer models were analyzed. Results : 37 articles were included. For producing animal cancer models BALB/C mice(n=22) and C57BL/6 mice(n=17) were selected. And intravenous injection of B16-F10 melanoma cells into tail vein(n=14) or intraperitoneal injection of sarcoma-180 cells(n=14) were frequently used to induce cancer. Various pharmacopunctures were injected into acupoints $CV_{12}(n=19)$, $ST_{36}(n=8)$, $BL_{18}(n=8)$ or peritoneal cavity(n=6), tumor site(n=2), tail vein(n=2). Outcome measures were categorized into anti-cancer, anti-metastasis, general condition, cytotoxicity, immune response, toxicity. Median Survival Time(MST) and increase of life span(ILS)(n=26) was frequently used for evaluating anti-cancer effects. And pulmonary colonization assay(n=13) was frequently used for evaluating anti-metastasis effects Conclusions : Based on these data, further research would be needed to ascertain the effectiveness of pharmacopuncture for treating cancer and broaden the range of clinical applications.

• Journal of Gastric Cancer
• /
• 제5권4호
• /
• pp.252-259
• /
• 2005

위에 발생하는 원발성 소세포암은 매우 드물며 예후는 좋지 않아 초기에 발견되어도 60% 이상이 1년 이내에 사망한다. 본원 외과에서 수술치료를 받은 위소세포암 첫 번째 증례는 수술소견상 복막전이 소견 등으로 근치적 수술이 불가능하여 위공장문합술을 시행하였다. 수술 후 etoposide, cisplatin화학요법을 시행하고 6개월 뒤에 찍은 CT촬영상 복막전이, 림프절전이가 악화되어 paclitaxel, cisplatin으로 약제변경 하였으나 수술 후 14개월째 사망하였다. 두 번째 증례는 내시경 조직검사상 위선암과 소세포암의 복합 소견을 보였으며 CT 촬영상 복강동맥주위 림프절종대 및 간전이 소견이 발견되었다. TS-1과 cisplatin 선행화학요법 2차 시행 후 림프절 종대는 완전관해, 원발소 및 간전이소는 부분관해 소견을 보여 위전절제술 및 확대림프절 절제술을 시행하였다. 수술로 절제된 위 및 주변 림프절 35개의 조직검사상 암세포가 모두 사멸되었으며 위내 원 발병소는 심한 심유화변성 소견을 보여 수술 전 사용한 항암요법이 유의했다고 판단되었다. 이에 수술 후에도 동일 제제로 4차례 추가 투약을 하였다. 수술 후 6개월에 시행한 CT촬영상 간전이가 진행된 소견을 보여 간우엽 후부절제술을 시행하고 이후 ininotecan과 cisplatin을 이용한 항암화학요법을 5차례 시행하고 있으며 술 후 14개월째 생존 중이다. 세 번째 증례는 순수 소세포암으로 근치적 위아전절제술을 시행하였으며 수술 후 5차례에 걸쳐 TS-1, cisplatin 보조항암화학요법 시행하였고 수술 후 13개월째 재발 없이 생존 중이다.

• Toxicological Research
• /
• 제15권1호
• /
• pp.79-87
• /
• 1999

To know the stress response and antioxidative effect of sulfur containing compounds, we observed the expression of the stress protein (heat shock protein; inducible protein) from mouse tissues and evaluated the protective effects to hydroxyl radical in mouse brain cell culture. Cysteine, methionine or sodium sulfide was fed by oral administration of 1 ml/per 6hr/three times with 1 mM, 2mM or 3mM to mouse, respectively. After that, the stress proteins were extracted from mouse tissues and analyzed the features of expression. The stress proteins by sulfur containing compounds were showed different aspects in the kinds and concentrations of their compounds, and in the tissues of mouse. In the liver, the stress proteins were appeared at different time on the concentration of sulfur containing compounds and had less than 20 KDa as small molecules. In general, the molecular weights of stress protein in liver, the stress proteins were appeared at different time on the concentration of sulfur containing compounds and had less than 20 KDa as small molecules. In general, the molecular weights of stress protein in the spleen were evaluated from 32KDa to 50KDA, and the induced times were relatively late at high concentration of cysteine, early at low concentration of methionine or sodium sulfide. The stress proteins in mouse muscle were detected mostly between 24hr after treatment of sulfur containing compounds. Their molecular weights were 15~24KDa. In the antioxidative effects of sulfur containing compounds to hydroxyl radical, cell viabilities were measured by 63.2% at 10 $\mu\textrm{M}$, 65.5% at 50 $\mu\textrm{M}$, 68.6% at 100 $\mu\textrm{M}$, 78.3% at 150 $\mu\textrm{M}$, or 83.0% at 200 $\mu\textrm{M}$ of cysteine, respectively. At addition of methionine, the cell viabilities were assessed as 58.1% at 10 $\mu\textrm{M}$, 62.8% at 50 $\mu\textrm{M}$, 75.7% at 100 $\mu\textrm{M}$, 78.6% at 150 $\mu\textrm{M}$, and 79.2% at 200 $\mu\textrm{M}$ after 4hrs exposure with 20mU/ml glucose oxidase (GO) system, while the numbers of live cells to hydroxyl radicals in treatment of sodium sulfide were showed 48.6% at 10 $\mu\textrm{M}$, 54.8% at 100 $\mu\textrm{M}$, 51.8% at 150 $\mu\textrm{M}$, and 51.6% at 200 $\mu\textrm{M}$ in the neuronal cells. In the inhibitory effects on the proliferation of tumor cells, percentages of dead cells of the CT-26 or HeLa cell were generally less than 30% even 48hr after addition of sulfur containing compounds. Conclusively, the results of these experiments indicate that stress protein by sulfur containing compounds can be used as physiological indicator for animal nutrition and for environment, and also that cysteine and methionine can play critical roles as an antioxidant.

• 생명과학회지
• /
• 제20권2호
• /
• pp.213-218
• /
• 2010

본 연구에서는 대장암 세포주 모델에서 파이토케미칼 capsaicin에 의한 항 생장 활성과 유전체 수준에서의 유전자 발현 변화를 연구하였다. 그 결과, 처리한 capsaicin 농도 의존적으로 세포 생존율이 감소함을 확인하였고, capsaicin은 다양한 유전자의 발현 변화를 유도하였다. DNA microarray 실험결과 $100\;{\mu}M$ capsaicin의 처리에 의해 2배 이상 증가되는 유전자 103개가 확인된 반면, 2배 이상 발현이 감소되는 유전자 153개가 확인되었다. 발현이 증가되는 유전자 중 4개(NAG-1, DDIT3, GADD45A 그리고 PCK2)를 선택하여 RT-PCR을 수행한 결과, DNA micorarray 실험과 일치함을 확인하였다. 또한 $100\;{\mu}M$ capsaicin의 처리에 의해 암 억제유전자인 p53의 발현이 증가됨을 RT-PCR과 real-time PCR 방법으로 확인하였다. 게다가, NAG-1, DDIT3 그리고 GADD45A 유전자는 p53의 존재에 관계없이 발현이 증가되는 반면, PCK2 유전자는 반드시 p53에 의해 발현이 유도됨을 확인할 수 있었다. 이러한 연구는 대장암 세포주에서 capsaicin에 의한 항암 기전을 이해하는데 도움을 줄 것으로 기대된다.

• 생명과학회지
• /
• 제30권6호
• /
• pp.501-512
• /
• 2020

제주조릿대 잎은 항염, 해열 및 이뇨작용을 가지고 있어 위궤양, 목마름 및 토혈 치료를 위한 민간의약으로 사용되어 왔다. 본 저자들은 제주조리대 잎에서 분리한 피토케미칼 풍부 추출물(PRE)과 그 에틸아세테이트 분획물(EPRE)은 여러 위암 세포주에서 세포사멸을 유도하는 항암 효과가 있다고 보고한 바 있다. 본 연구는 EPRE의 세포사멸 유도 기전에 관여하는 분자표적들을 탐색하기 위하여 EPRE을 처리한 SNU-16 세포에서 mRNA와 microRNA (miRNA)의 프로파일 변화를 분석하였다. RNA sequencing 분석을 통해 총 2,875개의 차등적으로 발현되는 유전자들(DEGs)을 동정하였다. 유전자 온톨로지(GO)와 KEGG 경로 분석 결과, EPRE는 세포사멸, 유사 분열-활성화 단백질 키나제(MAPK) 및 염증 반응, 종양 괴사 인자(TNF) 신호 전달 및 암 경로에 관여하는 유전자들의 발현을 조절하는 것으로 나타났다. 단백질-단백질 상호 작용(PPI) 네트워크 분석으로 세포사멸 및 세포죽음과 관련된 유전자들 간의 상호작용들을 확인할 수 있었다. 그리고, miRNA sequencing 분석을 통해 총 27개의 차별적으로 발현되는 miRNAs (DEMs)를 동정하였다. GO와 KEGG 경로 분석 결과, EPRE는 세포주기, 세포사멸 및 tropomyosin-receptor-kinase (TRK) 수용체 신호 전달, 성장인자-β(TGF-β), 핵인자 κB (NF-κB) 및 암 경로에 관여하는 miRNAs의 발현을 조정하였다. 본 연구결과는 EPRE의 항암 효과의 근본적인 메커니즘에 대한 통찰력을 제공한다.

• 생명과학회지
• /
• 제11권2호
• /
• pp.181-189
• /
• 2001

Cytokines are hormone-like proteins which mediate and regulast inflammatory and immune responses. Transforming growth factor -$\beta$1(TGF-$\beta$) plays an important role in the control of the immune response and wound healing, and in the development o various tissues and organs, Nitric oxide(NO) is major messenger molecule regulating immune function and blood vessel dilation and serving as a neurotransmitter in the brain and peripheral nervous system. Also, NO is to be a potent mutagen that cause mutation in the p53 tumor suppressor gene in early phases of human gastric carcinogenesis. The purpose of this study was to investigate the effect of Helicobacter phlori lystes, lipopolysaccharide (LPS), and Staphylococcus enterotoxin B(SEB) on production of TGF-$\beta$1 and NO by human fibroblasts. Primary cultured human fibroblasts were incubated with H. pylori lysates(Hp), LPs, SEB, Hp+LPS, Hp+SEB, Hp+LPS+SEB. Cultured supernatants that were collected at 24, 48 and 72 hr were assessed for TGF-$\beta$1 by enzyme-linked immunosorbent assay and NO production by quantification of nitrite ion. TGF-$\beta$1 production in fibroblasts exposed with Hp, LPS or SEB for 48 hrs was enhanced, but for 72 hrs inhibited. Its production by doble exposure such as Hp+LPS, Hp+SEB, Hp+LPS+SEB was lowered in comparison with single exposure of Hp in cases of 24 and 48 hrs incubation, but for 72 hrs decreased in Hp vaculoating toxin(+), increased in Hp vacuolating toxin(-). No production in fibroblasts increaed at all doses of LPS. But its production by exposure of SEB increased or decreased according to dose and incubation time. Also, NO production by Hp vacuolating toxin(+) increased at all doses, but its production by Hp vacuolating toxin(-) decreased. Its production by doble exposure such as Hp+LPS, Hp+SEB, Hp+LPS+SEB decreased in comparison with single exposure Hp Therefore, quantities pf TGB-$\beta$1 and NO released by human fibroblasts shows differences according to kinds of stimulants. Also, in care stimulated with same kinds of stimulants, its productions exhibit quantitative differences according to exposure times. These results suggest that the decreased of TGF-$\beta$1 in fibroblasts by mixed exposure with Hp producing vacuolating toxin and bacterial toxins such as LPS and SEB may effect negatively in healing of host tissue and increased of NO by infection oh H. pylori may related to the increased susceptibility for human gastric carcinogenesis.

• 생명과학회지
• /
• 제14권5호
• /
• pp.874-881
• /
• 2004

혈관 신생은 암의 성장 및 전이뿐만 아니라 염증, 관절염, 건성, 동맥경화 등의 병적인 진행에 주요한 역할을 하며, 혈관신생 억제를 통한 암의 치료를 시도하는 연구들이 활발하게 진행되고 있다. 혈관 신생 시 내피세포의 증식, 이동을 유도하는 활성화 과정이 필수적으로 일어나는 것으로 알려져 있다 본 연구에서는 in vitro에서 내피세포를 배양하여, 각종 growth factor가 풍부한 배지에서 활성화 시켰을 때, 그렇지 않는 세포들과의 유전자 발현 형태를 비교 조사하였다. HUVEC을 70∼80% cofluency로 배양시킨 후에 endothelial cell growth supplement (ECCS), 20% fetal bovine serum, heparin이 첨가된 Ml99 배지에서 13 시간 활성화시킨 세포(AHUVEC)와 대조군 세포(RHUVEC)로부터 분리한 total RNA로부터 CDNA를 제작하였고, 이것을 18,864 개의 유전자가 올려져있는 인간 올리고 칩과 hybridization 반응을 시켰다. 반응된 유전자를 이용하여 random clustering분석을 실시한 결과, 활성화 시켰던 HUVEC과 그렇지 않은 HUVEC으로 dendrogram 상에서 두개의 subgroup으로 나뉘어 지는 것을 확인할 수 있었다. 최소 2배 이상 발현 변화가 있는 유전자 122종이 활성화 시켰던 HUVEC으로부터 추출되었다. 이중에서 기능이 알려진 32 개의 유전자는 활성화시킨 HUVEC에서 발현이 증가하였고, 38 개의 유전자 발현은 감소하였다. 흥미롭게도 세포 증식과 이동, 염증, 면역반응에 관련한 유전자의 발현이 증가된 반면에 세포 흡착과 혈관 조직과 기능에 관련한 유전자의 발현이 감소된 것이 관찰되었다. 예상외로 규명이 잘된 혈관신생 인자와 관련한 유전자들의 발현에는 크기 차이를 보이지 않았으나, Eph-B4의 발현은 약 4 배 감소된 것으로 관찰되었다 또한, 2배 이상 발현에 차이를 보이고 기능이 알려져 있지 않은 유전자 52종이 발견되었다. 따라서, 이러한 연구 결과로부터 새로운 혈관 표적 물질 개발에 대한 기회가 제공될 수 있을 것이라 사료된다.