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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2017.1
- 수록면
- 230 - 245 (16page)
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Purpose The purpose of this study is to examine the role of the double-stranded RNA (dsRNA) activated Toll–interleukin-1 receptor domain-containing adaptor inducing interferon (TRIF) signal pathway in triggering apoptosis in hepatocellular carcinoma (HCC) cells.
Materials and Methods First, siRNA targeted autophagy–related gene LC3 (pU6H1-LC3 siRNA and siLC3) and a dsRNA used as a Toll-like receptor 3 (TLR3) ligand was constructed and synthesized, respectively. Then, a human HCC cell line was transfected with dsRNA, siLC3, and cotransfected with siLC3 and dsRNA (siLC3+dsRNA), respectively. Finally, quantification real-time polymerase chain reaction, western blotting, and immunofluorescence staining were used in the HCC line (SMMC7721), and MTT assay, flow cytometry, terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling, and transmission electron microscopy were used in an HCC xenograft model of nude mice. Human umbilical vein endothelial cell tube forming assay, color Doppler ultrasonographic flow image examination, and CD34-positive microvessel density were used in vitro and in vivo.
Results Compared with untreated cells, the protein and mRNA expression of TLR3 and TRIF was up-regulated, in order, siLC3+dsRNA, dsRNA, and siLC3. Expression of LC3 was obviously down-regulated and the autophagosomes were significantly decreased in siLC3+dsRNA and siLC3, whereas in dsRNA (p < 0.05). LC3 and TRIF colocation was observed in HepG2 cells. Decreased cell viability, increased apoptosis, decrease in xenograft tumor volume, and angiogenesis potential were also observed in order (p < 0.05).
Conclusion Suppression of intracellular autophagy resulted in decreased degradation of TRIF protein, which can promote triggering of apoptosis by the TLR3-TRIF pathway. dsRNA and siLC3 could play anticancer roles in coordination.
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참고문헌 (24)
참고문헌 신청
Amaravadi RK / 2011 / Principles and current strategies for targeting autophagy for cancer treatment / Clin Cancer Res 17 : 654 ~ 666
Biswas N / 2011 / The ubiquitin-like protein PLIC-1 or ubiquilin 1 inhibits TLR3-Trif signaling / PLoS One 6 : e21153 ~
Chen GY / 2012 / Simultaneous induction of autophagy and toll-like receptor signaling pathways by graphene oxide / Biomaterials 33 : 6559 ~ 6569
Chen L / 2012 / TLR3 dsRNA agonist inhibits growth and invasion of HepG2.2.15 HCC cells / Oncol Rep 28 : 200 ~ 206
Delgado MA / 2008 / Toll-like receptors control autophagy / EMBO J 27 : 1110 ~ 1121
Biswas N / 2011 / The ubiquitin-like protein PLIC-1 or ubiquilin 1 inhibits TLR3-Trif signaling / PLoS One 6 : e21153 ~
Chen GY / 2012 / Simultaneous induction of autophagy and toll-like receptor signaling pathways by graphene oxide / Biomaterials 33 : 6559 ~ 6569
Chen L / 2012 / TLR3 dsRNA agonist inhibits growth and invasion of HepG2.2.15 HCC cells / Oncol Rep 28 : 200 ~ 206
Delgado MA / 2008 / Toll-like receptors control autophagy / EMBO J 27 : 1110 ~ 1121
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