간암에서 Long noncoding RNA 프로파일링
Profiling of long non-coding RNA in hepatocellular carcinoma
- 주제(키워드) Hepatocellular carcinoma (HCC) , transcripts expression profiling , Long non-coding RNA (lncRNA) , antisense RNA
- 발행기관 아주대학교
- 지도교수 우현구
- 발행년도 2017
- 학위수여년월 2017. 8
- 학위명 박사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000025972
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Hepatocellular carcinoma (HCC) is the predominant subject of liver malignancies which provoke global concern. Previous studies have shown that long non-coding RNAs (lncRNAs) are differentially expressed in HCC, which have distinct functional roles in the pathogenesis of HCC. Recent advance of high-throughput RNA sequencing have revealed that numerous non-coding RNAs are expressed in cancers. Here, to deleineate functional lncRNAs in HCC, RNA-seq profiling was performed. I found 23 differentially expressed lncRNAs in the HCCs compared to non-tumoral tissues. Of these, 15 lncRNAs showed the correlated expression between antisense and sense lncRNAs, inclduding HAND2-AS1 and HAND2. The functional effects of HAND2-AS1 or HAND2 expression in liver cancers were further evaluated. Overexpression of HAND2-AS1 or HAND2 inhibited the proliferation and migration of liver cancer cells in vitro. The xenografted in vivo mouse model also showed significant reduction of tumor size by knockdown of HAND2. In addition, I found that the HAND2 mRNA expression was induced by HAND2-AS1 overexpression. This HAND2-AS1-mediated HAND2 expression was mediated throguht phosphorylation of CREB transcription factor. In summary, genome-wide noncoding RNA profiling analysis could identify that HAND2-AS1 can play pivotal roles in liver cancer progression.
more목차
I. INTORDUCTION 1
II. MATERIALS AND METHODS 3
1. Cell culture 3
2. Clinical samples 3
3. RNA isolation, DNA library preparation and sequencing 3
4. Gene expression constructs and lentiviral vector transfection 4
5. quantitative RT-PCR 5
6. Western Blotting 5
7. Colony formation assay 6
8. Migration assay 6
9. Xenograft Mouse model 7
10. Construction of luciferase reporter plasmid 7
11. Luciferase reporter assay 8
12. Estimation of transcript expression 8
13. Analysis of public data 9
14. Statistical analysis 9
III. RESULTS 10
1. Overall distribution of whole transcripts in hepatocellular carcinoma by RNA-seq 10
2. Differentially expressed antisense lncRNAs in HCC 15
3. Clinical implication of HAND2 and HAND2-AS1 18
4. Construction of HAND2 or HAND2-AS1 expressing cells 20
5. Tumor suppression by HAND2 or HAND2-AS1 in vitro 22
6. Tumor suppression by HAND2 in vitro 25
7. Tumor suppression by HAND2 or HAND2-AS1 in vivo 28
8. HAND2-AS1 induced HAND2 expression 30
IV. DISCUSSION 34
V. CONCLUSION 36
VI. REFERENCES 37
국문 요약 43
