RESUMO
SNPs in human AFP promoter are associated with serum AFP levels in hepatocellular carcinoma (HCC), suggesting that AFP promoter variants may generate better transcriptional activities while retaining high specificity to AFP-producing cells. We sequenced human AFP promoters, cloned 15 different genotype promoters and tested their reporter activities in AFP-producing and non-producing cells. Among various AFP variant fragments tested, EA4D exhibited the highest reporter activity and thus was selected for the further study. EA4D was fused with tBid and coupled with nano-particle vector (H1) to form pGL3-EA4D-tBid/H1. pGL3-EA4D-tBid/H1 could express a high level of tBid while retain the specificity to AFP-producing cells. In a HCC tumor model, application of pGL3-EA4D-tBid/H1 significantly inhibited the growth of AFP-producing-implanted tumors with minimal side-effects, but had no effect on non-AFP-producing tumors. Furthermore, pGL3-EA4D-tBid/H1 could significantly sensitize HCC cells to sorafenib, an approved anti-HCC agent. Collectively, pGL3-EA4D-tBid/H1, a construct with the AFP promoter EA4D and the novel H1 delivery system, can specifically target and effectively suppress the AFP-producing HCC. This new therapeutic tool shows little toxicity in vitro and in vivo and it should thus be safe for further clinical tests.
Assuntos
Proteína Agonista de Morte Celular de Domínio Interatuante com BH3/genética , Carcinoma Hepatocelular/terapia , Terapia Genética/métodos , Vetores Genéticos/administração & dosagem , Neoplasias Hepáticas/terapia , Nanopartículas , Regiões Promotoras Genéticas/genética , alfa-Fetoproteínas/genética , Carcinoma Hepatocelular/genética , Celulose/química , Ciclodextrinas/química , Ácido Fólico/química , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Nanopartículas/química , Polietilenoimina/química , Resultado do Tratamento , Células Tumorais CultivadasRESUMO
The aim of this study was to make a comparison between the tissue fusion technique and conventional methods for sealing bowel anastomosis. Eighteen female domestic pigs (Suidae, Sus) were used in our study. Tissue-fused anastomoses (LigaSure groups) were made in 13 animals (5 anastomoses per animal), which were subdivided into 4 groups according to different manufacturing settings: "LigaSure-L-1" and "LigaSure-L-2", with low energy output level with 1 or 2 device-activated tissue sealing times, and "LigaSure-M" and "LigaSure-H", with medium or high energy output level. As controls, automatically stapled (GIA group) and hand-sewn (suture group) anastomoses were utilized in 3 and 2 animals, respectively. There was no statistical difference in the overall leakage rate between the GIA group (6.7%) and the LigaSure groups (15%) (P=1.000). There was less proliferating epithelium covering the anastomosis gap in the LigaSure groups compared with the other two groups. The gap between the two extremities of muscular layers of the anastomosis in the LigaSure groups was filled with collagen fibers. More proliferating cell nuclear antigen (PCNA)-positive cells were found in the anastomoses of the LigaSure groups compared with the other two groups (P=0.010). Our results showed that the tissue fusion technology was a feasible and safe method for anastomoses.
Assuntos
Anastomose Cirúrgica/métodos , Intestino Delgado/cirurgia , Técnicas de Sutura , Engenharia Tecidual , Animais , Feminino , Modelos Animais , Reprodutibilidade dos Testes , SuínosRESUMO
The aim of this study was to make a comparison between the tissue fusion technique and conventional methods for sealing bowel anastomosis. Eighteen female domestic pigs (Suidae, Sus) were used in our study. Tissue-fused anastomoses (LigaSure groups) were made in 13 animals (5 anastomoses per animal), which were subdivided into 4 groups according to different manufacturing settings: "LigaSure-L-1" and "LigaSure-L-2", with low energy output level with 1 or 2 device-activated tissue sealing times, and "LigaSure-M" and "LigaSure-H", with medium or high energy output level. As controls, automatically stapled (GIA group) and hand-sewn (suture group) anastomoses were utilized in 3 and 2 animals, respectively. There was no statistical difference in the overall leakage rate between the GIA group (6.7%) and the LigaSure groups (15%) (P=1.000). There was less proliferating epithelium covering the anastomosis gap in the LigaSure groups compared with the other two groups. The gap between the two extremities of muscular layers of the anastomosis in the LigaSure groups was filled with collagen fibers. More proliferating cell nuclear antigen (PCNA)-positive cells were found in the anastomoses of the LigaSure groups compared with the other two groups (P=0.010). Our results showed that the tissue fusion technology was a feasible and safe method for anastomoses.