P-94 인간전분화능줄기세포 유래 폐상피전구세포의 카드뮴 세포독성 검사에서의 적용

허혜련,김지영,김우진,홍석호 대한결핵 및 호흡기학회 2016 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.121 No.0

인간전분화능줄기세포 (hPSC)는 중금속과 미세먼지와 같은 유해한 물질에 대한 독성 검사에 유용한 자원으로 여겨지고 있다. 또한 담배와 미세먼지의 공통 요소인 카드뮴을 사용하여 다양한 호흡기 질환의 병리학적 기전을 밝히고 예방을 위한 많은 연구가 진행되고 있다. 이전 연구에서 우리는 BEAS2B (human bronchial epithelial cell line)에서 카드뮴으로 인한 염증반응, 소포체 스트레스, 세포자살 관련 유전자변화 양상을 확인했다. 그래서 이번 연구에서는 인간줄기세포 유래 폐상피전구세포 (AETII, alveolar epithelial progenitor type II)에서도 카드뮴에 의해 유전자 그룹들의 변화가 유사하게 나타나는지를 확인하고자 했다. 순차적인 분화 방법을 통해 줄기세포를 AET II로 유도하고 FACS를 통해 NKX2.1, EpCAM, SpC, CPM의 표현형을 확인하였다. 24시간 동안 1, 5, 10 μM 농도의 카드뮴을 세포에 노출한 뒤 real time PCR를 통해 염증반응, 소포체 스트레스, 세포자살 관련 유전자의 발현을 확인했다. 이 연구는 줄기세포 유래 폐상피전구세포가 다양한 호흡기질환 및 재생의학에서 병리학적 기전을 이해하고 유해한 물질의 세포독성 평가를 할 수 있는 유용한 재료가 될 수 있음을 보여준다.

P-152 Cadmium-induced ER stress and inflammation are mediated through C/EBP-DDIT3 signaling in human bronchial epithelial cells

허혜련,김지영,송행석,김정운,김혜련,홍윤기,양세란,한선숙,이승준,김우진,홍석호 대한결핵 및 호흡기학회 2017 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.124 No.0

Cadmium (Cd), a major component of cigarette smoke, disrupts the normal functions of airway cells and can lead to the development of various pulmonary diseases such as chronic obstructive pulmonary disease (COPD). However, the molecular mechanisms involved in Cd-induced pulmonary diseases are poorly understood. Here, we identified a cluster of genes that are altered in response to Cd exposure in human bronchial epithelial cells (BEAS-2B) and demonstrated that Cd-induced ER stress and inflammation are mediated via CCAAT-enhancer-binding proteins (C/EBP)-DNA-damaged-inducible transcript 3 (DDIT3) signaling in BEAS-2B cells. Cd treatment led to marked upregulation and downregulation of genes associated with the cell cycle, apoptosis, oxidative stress and inflammation as well as various signal transduction pathways. Gene set enrichment analysis revealed that Cd treatment stimulated the C/EBP signaling pathway and induced transcriptional activation of its downstream target genes, including DDIT3. Suppression of DDIT3 expression using specific small interfering RNA effectively alleviated Cd-induced ER stress and inflammatory responses in both BEAS-2B and normal primary normal human bronchial epithelial cells. Taken together, these data suggest that C/EBP signaling may have a pivotal role in the early induction of ER stress and inflammatory responses by Cd exposure and could be a molecular target for Cd-induced pulmonary disease. This work was supported by NRF(National Research Foundation of Korea) Grant funded by Korean Government (2016H1A2A1909769-Global Ph.D. Fellowship program)

Isolated Star Formation in DC303.8-14.2

허혜련,이정은 한국천문학회 2008 天文學會報 Vol.33 No.2

P-73 MicroRNA Profiling Reveals a Role for miR-181α-2-3p in the Pathogenesis of Chronic Obstructive Pulmonary Disease

김지영,허혜련,홍석호,김우진 대한결핵 및 호흡기학회 2016 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.121 No.-

Chronic obstructive pulmonary disease (COPD) is a complex disorder that is characterized by airflow limitation associated with chronic inflammation. MicroRNA (miRNAs) are one of the main types of epigenetic factor, which can act as key regulators of gene expression, modulate almost all biological process. Despite the fact that expression changes of several miRNAs play an important role in COPD, little is known about the role of miRNAs in inflammatory disorders. In this study, we aimed to identify dysregulated miRNAs in serum samples of COPD patients and the underlying molecular mechanism. Among the top 20 differentially expressed miRNAs, five were also differentially expressed in serum samples from COPD subjects versus those from control subjects. Specifically, miR-181a-2-3p was down-regulated significantly in the COPD samples, whereas miR-181d-5p, miR-501-3p, miR-769-5p, and miR-191-5p were up-regulated significantly. Based on the results of the miRNA profiling, we further validated that the expression of miR181a-2-3p was also decreased in bronchial epithelial cells treated with Cd in vitro. Interestingly, bioinformatics analysis demonstrated that high mobility group box 1(HMGB1) an important inflammatory mediator of COPD experimental model, were predicted as a target of miR-181a-2-3p, which was validated by qPCR and western blotting. Furthermore, anti-miR-181a-2-3p significantly enhanced mitochondrial dysfunctions proteins and MAPK signaling pathway. Taken together, these data suggest that miR181a-2-3p can regulate inflammatory responses by targeting HMGB1

Cadmium-induced ER stress and inflammation are mediated through C/EBP–DDIT3 signaling in human bronchial epithelial cells

김지영,송행석,허혜련,김정운,김혜련,홍윤기,양세란,한선숙,이승준,김우진,홍석호 생화학분자생물학회 2017 Experimental and molecular medicine Vol.49 No.-

Cadmium (Cd), a major component of cigarette smoke, disrupts the normal functions of airway cells and can lead to the development of various pulmonary diseases such as chronic obstructive pulmonary disease (COPD). However, the molecular mechanisms involved in Cd-induced pulmonary diseases are poorly understood. Here, we identified a cluster of genes that are altered in response to Cd exposure in human bronchial epithelial cells (BEAS-2B) and demonstrated that Cd-induced ER stress and inflammation are mediated via CCAAT-enhancer-binding proteins (C/EBP)-DNA-damaged-inducible transcript 3 (DDIT3) signaling in BEAS-2B cells. Cd treatment led to marked upregulation and downregulation of genes associated with the cell cycle, apoptosis, oxidative stress and inflammation as well as various signal transduction pathways. Gene set enrichment analysis revealed that Cd treatment stimulated the C/EBP signaling pathway and induced transcriptional activation of its downstream target genes, including DDIT3. Suppression of DDIT3 expression using specific small interfering RNA effectively alleviated Cd-induced ER stress and inflammatory responses in both BEAS-2B and normal primary normal human bronchial epithelial cells. Taken together, these data suggest that C/EBP signaling may have a pivotal role in the early induction of ER stress and inflammatory responses by Cd exposure and could be a molecular target for Cd-induced pulmonary disease.

Inhibition of MicroRNA-221 and 222 Enhances Hematopoietic Differentiation from Human Pluripotent Stem Cells via c-KIT Upregulation

이지윤,김명주,허혜련,하권수,한은택,박원선,양세란,홍석호 한국분자세포생물학회 2018 Molecules and cells Vol.41 No.11

The stem cell factor (SCF)/c-KIT axis plays an important role in the hematopoietic differentiation of human pluripotent stem cells (hPSCs), but its regulatory mechanisms involving microRNAs (miRs) are not fully elucidated. Here, we demonstrated that supplementation with SCF increases the hematopoietic differentiation of hPSCs via the interaction with its receptor tyrosine kinase c-KIT, which is modulated by miR-221 and miR-222. c-KIT is comparably expressed in undifferentiated human embryonic and induced pluripotent stem cells. The inhibition of SCF signaling via treatment with a c-KIT antagonist (imatinib) during hPSC-derived hematopoiesis resulted in reductions in the yield and multi-lineage potential of hematopoietic progenitors. We found that the transcript levels of miR-221 and miR-222 targeting c-KIT were significantly lower in the pluripotent state than they were in terminally differentiated somatic cells. Furthermore, suppression of miR-221 and miR-222 in undifferentiated hPSC cultures induced more hematopoiesis by increasing c-KIT expression. Collectively, our data implied that the modulation of c-KIT by miRs may provide further potential strategies to expedite the generation of functional blood cells for therapeutic approaches and the study of the cellular machinery related to hematologic malignant diseases such as leukemia.

ACN9 Regulates the Inflammatory Responses in Human Bronchial Epithelial Cells

정재훈,김지영,김정운,허혜련,정진선,류영준,홍윤기,한선숙,홍석호,이승준,김우진 대한결핵및호흡기학회 2017 Tuberculosis and Respiratory Diseases Vol.80 No.3

Background: Airway epithelial cells are the first line of defense, against pathogens and environmental pollutants, in the lungs. Cellular stress by cadmium (Cd), resulting in airway inflammation, is assumed to be directly involved in tissue injury, linked to the development of lung cancer, and chronic obstructive pulmonary disease (COPD). We had earlier shown that ACN9 (chromosome 7q21), is a potential candidate gene for COPD, and identified significant interaction with smoking, based on genetic studies. However, the role of ACN9 in the inflammatory response, in the airway cells, has not yet been reported. Methods: We first checked the anatomical distribution of ACN9 in lung tissues, using mRNA in situ hybridization, and immunohistochemistry. Gene expression profiling in bronchial epithelial cells (BEAS-2B), was performed, after silencing ACN9 . We further tested the roles of ACN9, in the intracellular mechanism, leading to Cd-induced production, of proinflammatory cytokines in BEAS-2B. Results: ACN9 was localized in lymphoid, and epithelial cells, of human lung tissues. ACN9 silencing, led to differential expression of 216 genes. Pathways of sensory perception to chemical stimuli, and cell surface receptor-linked signal transduction, were significantly enriched. ACN9 silencing, further increased the expression of proinflammatory cytokines, in BEAS-2B after Cd exposure. Conclusion: Our findings suggest, that ACN9 may have a role, in the inflammatory response in the airway.

바이오의약품 품질 확보를 위한 국가표준품의 안정성 평가

강민수,박태준,우희원,박은혜,김지화,허혜련,이철현,전형옥,류승렬 한국에프디시규제과학회(구 한국에프디시법제학회) 2022 FDC법제연구 Vol.17 No.2