간암전사체 분석을 통한 간암진행 조절 융합유전자의 발굴
Identification of fusion genes regulating liver cancer progression through transcript analysis
- 주제(키워드) Hepatocellular carcinoma , fusion gene , RNA-sequencing , SLC39A14 , PIWIL2
- 발행기관 아주대학교 일반대학원
- 지도교수 우현구
- 발행년도 2019
- 학위수여년월 2019. 2
- 학위명 박사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000028820
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Although Fusion genes have been recognized as an oncogene that affects development and progression of cancer, there are rarely reports of fusion genes in hepatocellular carcinoma (HCC), the third leading cause of cancer mortality worldwide. Advances of RNA sequencing (RNA-Seq) technology offer the ability to screen fusion genes. The current study aimed to determine whether fusion genes are present and affect cancer progression in HCC. Here, the fusion transcripts were identified by using the fusion detection pipeline based on the three different fusion-finder algorithms: SOAPfuse, ChimeraScan, and TopHat-Fusion. Out of 14 potential fusion transcripts that were not previously reported, SLC39A14-PIWIL2 was predicted as a potential oncogenic fusion gene through evaluating the expression of 3’ transcripts. SLC39A14, a metal-ion transporter, is known to maintain metal ion homeostasis by excreting zinc, iron, and manganese in the liver and pancreas, and PIWIL2 has been reported to causes tumorigenesis in organs other than testis. The structure of SLC39A14-PIWIL2 was formed by fusion of SLC39A14 exon 1 and PIWIL2 exons 7–23 on the same chromosome 8 through chromosomal inversion in one HCC patient. The truncated form of PIWIL2 was transcripted by SLC39A14 containing the predictive promoter region from SLC39A14-PIWIL2. Using overexpression system, it was demonstrated that SLC39A14-PIWIL2, wild type PIWIL2 (WT PIWIL2), and truncated PIWIL2 (tPIWIL2) were shown to affect proliferation, colony formation, migration, and invasion in the HCC cell lines (i.e., HepG2 and Huh7). In conclusion, these results suggest that SLC39A14-PIWIL2 fusion transcript identified by the fusion detection pipeline is a novel SLC39A14-PIWIL2 variant that is expected to play oncogenic roles by expressing truncated form of PIWIL2 in HCC.
more목차
I. INTRODUCTION 1
II. MATERIALS AND METHODS 9
1. Patients and tissue specimens 9
2. RNA-seq and novel transcripts profiling 9
3. Detection of fusion transcripts 10
4. Cell culture 10
5. Construction of expression vectors 10
6. Quantitative real-time PCR (qRT-PCR) and RT-PCR 11
7. Proliferation and colony formation assay 11
8. Migration and invasion assay 12
9. Statistical analysis 12
III. RESULTS 14
A. Identification of fusion genes in HCC 14
B. Characterization of SLC39A14-PIWIL2 18
C. tPIWIL2 contributes to the development and progression of HCC cell lines 26
IV. DISCUSSION 30
V. CONCLUSION 33
VI. REFERENCES 34
국문 요약 40
