Different microRNA expressions in gastric cancer depending on Helicobacter pylori infection

학위논문 (석사)-- 서울대학교 대학원 : 의과학과, 2014. 2. 김나영.Background and aim: Helicobacter pylori (H. pylori) infection increases the risk of gastric cancer through inducing aberrant gene expression regulation of cell. The microRNA (miRNA) regulate downstream target gene which can control cell proliferation and differentiation. The expressions of miRNA are usually analyzed with microarray and FFPE (formalin fixed paraffin embedded) sample can be used for microarray demanding high quality RNA. The study was undertaken to identify microRNAs differently expressed by H. pylori infection in patients with intestinal type of gastric cancer using miRNA microarray, and to confirm the candidate miRNAs expression levels. Methods: Total RNA was extracted from cancerous region and non-cancerous regions in formalin fixed paraffin embedded tissues of intestinal type gastric cancer patients who were H. pylori-positive (n=8) or -negative (n=8). The RNA was analyzed with a 3,523 miRNA profiling microarray based on the Sanger miRBase. Validation analysis was performed using TaqMan miRNA assays. Results: 219 miRNAs in the aberrant miRNA profiles across the miRNA microarray showed at least two-fold changed different expression in H. pylori-positive and -negative cancer tissue. Seven statistically significant candidate miRNAs were selected with online miRNA databasesmiRWalk and HMDD. TaqMan miRNA assays confirmed that three miRNAs (miR-99b-3p, miR-564 and miR-638) were significantly increased in H. pylori-positive cancer than H. pylori-negative. In addition, four miRNAs (miR-204-5p, miR-338-5p, miR-375 and miR-548c-3p) were significantly increased in H. pylori-negative cancer than H. pylori-positive. Conclusion: The miRNA expression in the intestinal type of gastric cancer depending on H. pylori infection suggest that different gastric cancer pathogenesis could be exist between H. pylori-positive and -negative gastric cancer. FFPE specimens can be used for investigating the miRNA expression patterns. Further study which focused on miRNA function in gastric carcinogenesis is needed.초록 ……………………………………………………………….ⅰ 목차 ……………………………………………………………….ⅲ LIST OF TABLES ……………………………………………..…..ⅳ LIST OF FIGURES ……………………………………………..…ⅴ 본문 ……………………………………………………………..… 1 서론 ………………………………………………………..…1 2 연구재료 및 방법 …………………………………………..2 3 연구결과 ………………………………………………..……7 4 고찰 ……………………………………………………..…..11 참고문헌 ………………………………………………………....20 국문초록 …………………………………………………………25Maste

Coronary Artery Intervention after Cytostatics Treatment in Unstable Angina Patient with Essential Thrombocythemia. A Case Report and Literature Review

Essential thrombocythemia (ET) is a clonal disorder of myeloid stem cells that causes thrombocytosis. As a result, ET can lead to vascular thrombosis and tissue ischemia; the association of coronary artery abnormalities such as myocardial infarction or unstable angina is rare. Here we describe a 45-year-old male patient with essential thrombocythemia who presented with unstable angina. Elective coronary angiography showed total occlusion of mid right coronary artery and mid left anterior descending coronary artery. ET was confirmed by a bone marrow biopsy; treatment was started with antiplatelet therapy including aspirin and clopidogrel along with cytostatic therapy with hydroxyurea and anagrelide. After the initiation of the treatment, the platelet count decreased to 20×104/µL. In addition, percutaneous coronary angioplasty was successfully performed with stent placement at the right coronary artery without hemorrhagic or thrombotic complications.ope

Sclerosing Encapsulating Peritonitis (Abdominal Cocoon) after Abdominal Hysterectomy

Sclerosing encapsulating peritonitis (SEP) is a poorly understood and rarely documented cause of small bowel obstruction. Although recurrent peritonitis has been reported as the main contributory factor leading to secondary SEP, the pathogenesis of primary (idiopathic) SEP is still uncertain. A 40-year-old woman with a history of total abdominal hysterectomy due to gestational trophoblastic disease presented with progressive lower abdominal pain and abdominal distension. Ultrasonography and contrast-enhanced abdomen-pelvis computed tomography of the abdomen revealed encapsulation of the entire small bowel with a sclerotic capsule. At laparotomy, a fibrous thick capsule encasing small bowel loops was revealed. Extensive adhesiolysis and removal of the capsule from the bowel loops were performed. The patient recovered uneventfully; she was discharged without complications. SEP is a rare cause of small bowel obstruction. We treated a case of abdominal cocoon with intestinal partial obstruction in a woman with a history of abdominal hysterectomy due to gestational trophoblastic disease. Surgical treatment was effective and the patient recovered without complication.ope

Association of the ABCB1 gene polymorphisms 2677G>T/A and 3435C>T with clinical outcomes of paclitaxel monotherapy in metastatic breast cancer patients

BACKGROUND: ABCB1 is responsible for multidrug resistance, the principal mechanism by which many cancers develop resistance to chemotherapeutic drugs. There is a controversy whether ABCB1 gene polymorphisms correlate with survival and response in cancer patients treated with chemotherapy. We evaluated the association between clinical outcome (safety and efficacy) of paclitaxel monotherapy in metastatic breast cancer patients with ABCB1 gene polymorphisms 2677G>T/A or 3435C>T. PATIENTS AND METHODS: Patients with metastatic breast cancer were treated with 175 mg/m(2) paclitaxel per 3-week cycle. Peripheral blood mononuclear cells from patients were used to genotype ABCB1 2677G>T/A and 3435C>T polymorphisms. Genotypes were investigated for their association with tumor response, survival, toxicity, and chemoresistance. RESULTS: ABCB1 3435 CT showed a significantly lower disease control rate than the CC genotype (P = 0.025). ABCB1 3435 CT was correlated with shorter overall survival (OS) in Cox regression analysis (P = 0.026). The 2677 GG genotype showed a significant association with chemoresistance to paclitaxel and anthracycline (P = 0.04 and 0.04, respectively). None of the ABCB1 genotypes correlated with toxicity. CONCLUSIONS: ABCB1 genotypes may be a predictor of paclitaxel activity as well as a prognostic factor in metastatic breast cancer patientsope

Adenosine triphosphate-based chemotherapy response assay (ATP-CRA)-guided versus empirical chemotherapy in unresectable non-small cell lung cancer

BACKGROUND: We retrospectively compared adenosine triphosphate-based chemotherapy response assay (ATP-CRA)-guided and empirical chemotherapies for unresectable non-small cell lung cancer (NSCLC) in this case-control study. PATIENTS AND METHODS: Unresectable NSCLC patients receiving ATP-CRA-guided platinum-based doublets as first-line therapy were enrolled as cases (n=27; 14 platinum-sensitive and 13 platinum-resistant patients). Performance status, stage, and chemotherapeutic regimen-matched patients receiving empirical chemotherapy were selected from the retrospective database as controls (n=93) in a case to control ratio of approximately 1:3. RESULTS: Response rate and survival (progression-free; overall) in both groups were not significantly different. However, the platinum-sensitive subgroup by ATP-CRA showed a higher response rate than the empirical group (71 versus 38%; p=0.023) with a trend toward longer progression-free survival (8.7 versus 4.8 months for platinum-sensitive versus empirical; p=0.223) and overall survival (not reached versus 12.6 months for platinum-sensitive versus empirical for p=0.134). CONCLUSION: ATP-CRA may be helpful in selecting platinum-responsive patients in unresectable NSCLC. We consider that nonplatinum doublets in platinum-resistant patients by ATP-CRA may be a more adapted approach than platinum-based doublets in future clinical trialsope

Concurrent chemoradiotherapy with 5FU/DDP in esophageal cancer patients

의학과/석사[한글]식도암에서 항암제-방사선 동시치료는 의학적으로 수술을 견디기 어려운 환자나 기술적으로 수술이 어려운 경우에 기본 치료로 고려되는 방법이다. 본 연구는 이를 바탕으로 연세 암센터에서 2000년부터 2005년까지 국소 진행성 식도암으로 진단받고 항암제-방사선 동시치료를 받은 29명의 환자들을 대상으로 치료 효과에 대해 후향적 연구분석을 하였다. 항암제치료로 cisplatin (DDP)은 60 mg/m2을 제 2일과 30일에 정주하였고 5-fluorouracil (5-FU)은 750 mg/m2을 제 1일에서 5일까지 그리고 29일에서 33일까지 점적 정주하였다. 항암제 치료와 함께 방사선 치료는 63 Gy를 하루에 1.8 Gy씩 주 5회씩 조사하여 총 7주간 시행하였다. 이후 항암제 유지요법으로 5-FU 1000 mg/m2을 각 주기의 제 1일에서 5일까지, DDP 80 mg/m2을 각 주기의 제 2일에 투여하였고 각 주기는 4주마다 반복하여 최대 4회까지 시행하였다.대상 환자는 남자 28예, 여자 1예로 총 29예이었으며 중간 연령은 64세 (범위: 45-77세)이었다. 환자들의 수행상태는 ECOG 기준으로 하여 29예 모두 1이하로 양호하였다. 임상병기는 II기가 2예, III기가 21예, 그리고 IV기 6예이었다. 대상환자에서 항암제-방사선 동시치료 후 항암제 유지요법 후의 전체 반응율은 58.6%이었다. 대상 환자의 중앙 추적기간은 32.5개월이었으며 중앙 무진행생존기간은 11.7개월, 중앙 생존기간은 17.7개월, 그리고 전체 2년 생존율은 39.6%이었다. 항암제-방사선 동시치료 중 NCI-CTC version 2.0 기준에 따른 Grade 3이상의 치료독성은 중성구 감소증, 구역•구토, 식도염이 각각 전체 환자의 20.6%, 20.6% 그리고 20.6%에서 보였다. 항암제 유지요법 중 Grade 3이상의 치료독성은 중성구 감소증, 구역•구토가 각각 전체 환자의 37.5%, 12.5%에서 보였다. 결론적으로 식도암 환자를 대상으로 DDP와 5-FU를 이용한 항암제-방사선 동시치료 후 항암제 유지요법을 시행한 본 연구의 결과는 다른 연구결과들과 유사한 치료성적을 보여 수술이 어려운 환자들을 대상으로 추천할 만한 치료방법으로 사료된다. 그러나 식도암의 치료효율을 더욱 개선을 위해서는 국소재발을 억제하기 위한 새로운 치료 방안이 개발되어야 할 것으로 생각된다. [영문]Concurrent chemoradiotherapy is considered the standard of care for patients with medically inoperable or surgically unresectable esophageal cancer.Therefore we performed a retrospective analysis to evaluate the efficacy and toxicity of concurrent chemoradiotherapy for 29 patients with locally advanced esophageal cancer who were treated at Yonsei cancer center from 2000 to 2005. Cisplatin (DDP) 60 mg/m2 was administered on days 2 and 30, and 5-fluorouracil (5-FU) 750 mg/m2 was administered on days 1-5 and 29-33 during radiotherapy. Concurrent radiation therapy included 1.8 Gy in 35 fractions over 7 weeks, for a total dose of 63 Gy. Maintenance chemotherapy after concurrent chemoradiotherpy was consisted of infusional 5-FU 1000 mg/m2 on day 1 to 5 and infusion of DDP 80 mg/m2 on day 2, every 4 weeks for four courses.Of the 29 patients, 28 patients were males and 1 female with median age of 64 years (range, 45-77). All patients had good performance status ( ECOG ≤ 1). Two patients were in stage II, 21 in III, and 6 in IV. The overall response rate, which encompassed complete and partial response, after concurrent chemoradiotherapy and maintenance chemotherapy was 58.6%. With a median follow–up duration of 32.5 months, median time to progression and overall survival was 11.7 months and 17.7 months, respectively. Overall 2 year survival rate of all patients was 39.6%. The toxicities exceeding grade 3 by NCI-CTC version 2.0 during chemoradiotherapy were found to be neutropenia, emesis and esophagitis in 20.6%, 20.6% and 20.6% of the 29 patients, respectively. Grade 3 or 4 neutropenia and emesis during maintenance chemotherapy were seen in 37.5% and 12.5% of the 24 patients, respectively.This study with concurrent chemoradiotherapy followed by maintenance chemotherapy with DDP and 5-FU showed comparable efficacies to previous reports in locally advanced esophageal cancer. However further studies are needed to overcome high local failure rate.ope

Gene expression signature-based prediction model for synergistic effect of histone deacetylase inhibitor and chemotherapy

Dept. of Medicine/박사Purpose We evaluated the cytotoxic effects of combining of suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, with taxanes in human gastric cancer cell lines, and studied the pre-treatment difference of gene profile to identify synergism-related genes that could potentially mediate the cytotoxic response. Next we developed predicting model for the chemosensitivity of taxane/SAHA combination. To understand the roles of synergism-related genes we investigated the change of protein expression related with cell cycle and HDAC. We also analyzed genetic variants by whole exome-sequencing to identify possible mechanisms that may be responsible for synergism. Methods Twenty five gastric cancer cell lines with 22K gene expression data were treated with SAHA and paclitaxel or docetaxel, and the synergistic interaction between the drugs was evaluated in vitro using the combination index (CI) method. We performed significance analysis of microarray (SAM) to identify chemosensitivity-related genes in gastric cancer cell lines that were concomitantly treated with SAHA and taxane. We generated a correlation-matrix between gene expression and CI values to identify genes whose expression correlated with a combined effect of taxanes and SAHA. On the basis of expression profiles of genes selected from microarray, we developed ‘Molecular Diagnosis Score (MDS)’ prediction model with quantitative RT-PCR (qRT-PCR). To identify the mechanism of synergism, mRNA expression of selected genes with RT-PCR, protein expression of cell cycle-, apoptosis-related genes, histone acetylation with western blot and cell cycle analysis with flow cytometry were investigated. We performed whole-exome sequencing on 40 gastric cancer cells using Illumina HiSeqTM. The sequencing data were examined for genes which were involved in histone acetylation, cell cycle regulation and tubulinsResults Taxane and SAHA combination had a synergistic cytotoxic effect against taxane-resistant gastric cancer cells. We selected 49 chemosensitivity-related genes, which were commonly identified in paclitaxel and docetaxel combined with SAHA, via SAM analysis. Among them, nine common genes were extracted from the subsequent correlation-matrix analysis. These genes were validated with qRT-PCR and used for developing MDS. Application of MDS scoring system for evaluating tumor tissue of gastric cancer patients who were treated with taxane predicted the sensitivity of taxane with 100% sensitivity, 43% specificity. The selected nine genes of gastric cancer cells were modulated by SAHA and taxane. The protein expression of p21 and acetylated histone H3 were significantly increased with paclitaxel/SAHA in synergism-inducing cell line. The frequency of OR4L1Gly109Ser, GBP4G2366R, KCNN3L66H , IGSF9G34E mutated cells was significantly higher in synergism-induced cells than other cells, respectively. HDAC9G366V mutation was exclusively identified in synergism- induced YCC-16. Conclusions Collectively, we discovered biomarkers for predicting chemosensitivity of SAHA/taxane combination to further aid the development of personalized chemotherapy regimen for gastric cancer patients. Prospective study is needed to fully define the clinical utility of the biomarkers.prohibitio

METTL16의 Notch 전사 조절에 따른 간암세포 이동 및 침습 영향

MasterⅠ. 서 론 1 1.1. 간세포암종 발달과 치료를 위한 전략 1 1.2. METTL16 과 암의 예후 2 1.3. Notch 신호 및 간세포암종에서 예후 4 1.4.원발성간암에서 METTL16와 Notch에 상관관계 5 Ⅱ. 실험재료 및 방법 6 2.1. 세포주 배양 6 2.2. 유전자 발현 벡터 6 2.3. 시약 6 2.4. 일시적인 형질주입 7 2.5. 실시간 중합효소 연쇄반응 7 2.6. 세포 증식 측정 (Cell proliferation assay) 9 2.7. 세포의 침습성 측정 (cell migration assay) 9 2.8. 세포의 이동성 측정(cell invasion assay) 9 2.9. 통계적 분석 10 Ⅲ. 결과 11 3.1. METTL16의 억제는 간암세포에서 NOTCH1의 발현을 증가시킨다. 11 3.2. METTL16의 억제는 간암세포에서 침습성과 이동성에 영향을 미친다. 17 Ⅳ. 고찰 및 결론 24 참고문헌 26 영문요약 2

Identification of genes related to a synergistic effect of taxane and suberoylanilide hydroxamic acid combination treatment in gastric cancer cells

PURPOSE: We evaluated the cytotoxic effects of combining suberoylanilide hydroxamic acid (SAHA), a histone deacetylase inhibitor, with taxanes in human gastric cancer cell lines and assessed the pre-treatment difference of gene expression to identify genes that could potentially mediate the cytotoxic response. METHODS: Gastric cancer cell lines were treated with SAHA and paclitaxel or docetaxel, and the synergistic interaction between the drugs was evaluated in vitro using the combination index (CI) method. We performed significance analysis of microarray (SAM) to identify chemosensitivity-related genes in gastric cancer cell lines that were concomitantly treated with SAHA and taxane. We generated a correlation matrix between gene expression and CI values to identify genes whose expression correlated with a combined effect of taxanes and SAHA. RESULTS: Combination treatment with taxane and SAHA had a synergistic cytotoxic effect against taxane-resistant gastric cancer cells. We identified 49 chemosensitivity-related genes via SAM analysis. Among them, nine common genes (SLIT2, REEP2, EFEMP2, CDC42SE1, FSD1, POU1F1, ZNF79, ETNK1, and DOCK5) were extracted from the subsequent correlation matrix analysis. CONCLUSIONS: The combination of taxane and SAHA could be efficacious for the treatment of gastric cancer. The genes that were related to the synergistic response to taxane and SAHA could serve as surrogate biomarkers to predict the therapeutic response in gastric cancer patients.ope