2026-09-22T05:02:54Zhttps://repository.ajou.ac.kr/oai/request

oai:repository.ajou.ac.kr:201003/75322024-10-11T02:17:40Zcom_201003_14372com_201003_14368col_201003_14373

Regulation of mRNA expression through 3’ UTR sequences of IL6 gene in amino acid-deprived HeLa cells HeLa 세포주의 아미노산 결핍에서 IL6 유전자의 3’ UTR이 mRNA의 표현에 미치는 영향분석 강, 정희 최, 용준 대학원 의생명과학과 201024346 강, 정희 ER stress responses and autophagic responses triggered by amino acid-deprivation of cancer cells activate STAT3 and NF-κB transcriptional factors subsequently to induce IL6 cytokines. Here, it was investigated whether 3’UTR of IL6 mRNA played a role in IL6 induction through cellular stress responses. In the studies with GFP-reporter containing 3’UTR of IL6 mRNA, it was elucidated that 3’UTR contributed to increase of IL6 mRNA after amino acid-starvation, and its motifs seemed to be broadly distributed according to deletion mutants studies of IL6 3’UTR. NF-κB functioned in 3’UTR to partially contribute to stabilization of IL6 mRNA according to knock-down experiments of p65, NF-κB subunit. Finally, treatments of p38 MAP kinase inhibitor, SB203580 markedly blocked mRNA stabilization through 3’UTR as well as IL6 induction after amino acid-starvation, suggesting that p38-MK2/3-TTP pathway is evidently activated and implicated in stabilization of IL6 mRNA during starvation of cancer cells. Conclusively, these studies indicate that stress signals via NF-κB and p38 lead to mRNA stabilization through 3’UTR sequences, and their molecular mechanism would need to be further studied. ABSTRACT ⅰ TABLE OF CONTENTS ⅱ LIST OF FIGURES ⅳ LIST OF TABLES ⅴ ABBREVIATION ⅴ Ⅰ. INTRODUCTION 1 Ⅱ. MATERIALS AND METHODS 3 A. Cell, antibodies and other reagents 3 B. Expression constructs and lentiviral transfections 3 C. Real time RT PCR 4 D. Western blot 4 Ⅲ. RESULTS 6 A. IL6 induction in amino acid-deprived HeLa cell 6 B. Constructions of GFP-reporter plasmids containing 3’UTR of IL6 mRNA 6 C. Effects of 3’UTR on induction of IL6 reporter in amino acids-deprived HeLa cells 8 D. Effect of partial 3’UTR on induction of IL6 reporter in amino acids-deprived HeLa cell 8 E.GFP expressions in HeLa cells infected with pCMV or pCMV-UTR before/after amino acid deprivation 8 F. Effects of STAT3 and NF-kB transcriptional factors on IL6 3’UTR in IL6 G. Effects of MAP kinase inhibitor on IL6 3’ UTR in amino acids-deprived HeLa cells 14 Ⅳ. DISCUSSION 19 Ⅴ. CONCLUSION 21 REFERENCES 22 국문요약 26 Master 2012-10-25T05:14:06Z 2012-10-25T05:14:06Z 2012 2012 Thesis Theses http://repository.ajou.ac.kr/handle/201003/7532 http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000012350 000000012350 en