HeLa 세포주의 아미노산 결핍에서 IL6 유전자의 3’ UTR이 mRNA의 표현에 미치는 영향분석
Regulation of mRNA expression through 3’ UTR sequences of IL6 gene in amino acid-deprived HeLa cells
- 발행기관 아주대학교
- 지도교수 최 용준
- 발행년도 2012
- 학위수여년월 2012. 2
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000012350
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
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.
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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 induction of amino acid deprived
HeLa cells 14
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
