Multi-target lentivirus specific to hepatocellular carcinoma: In vitro and in vivo studies
| Autor: | Jian Niu, He-wei Zhao, Jingfeng Sun, Ye-wei Zhang, JianFei Wen, Wan Yee Lau, Ji-feng Feng, Xiang Lu, Jindao Wu, Xia He, Dong-Liang Yan, Huan-zhou Xue, Guo-Wen Yin, Yin-xue Yang |
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| Rok vydání: | 2013 |
| Předmět: |
Male
Carcinoma Hepatocellular Apoptosis Biology Receptor IGF Type 1 Mice In vivo medicine Animals Humans RNA Small Interfering Mice Inbred BALB C TUNEL assay Hepatology Cell growth Lentivirus Liver Neoplasms Genetic Therapy medicine.disease biology.organism_classification Molecular biology MicroRNAs Terminal deoxynucleotidyl transferase Hepatocellular carcinoma Female alpha-Fetoproteins Liver cancer |
| Zdroj: | Journal of Hepatology. 58:502-508 |
| ISSN: | 0168-8278 |
| Popis: | Background & Aims We aimed at investigating the effects of the targeted transduction of the Wtp53-pPRIME-miR30-shRNA gene into liver cancer cells, under the mediation of anti-alpha fetoprotein scFv-directed lentivirus, and the inhibitory effect of this system on liver cancer cells. Methods The result of infection was observed by fluorescence microscopy. Polymerase chain reaction and Western blotting were used to demonstrate the successful transduction and transcription of the Wtp53-pPRIME-miR30-shRNA-IGF1R gene. Cell growth was observed via the Cell-Counting Kit-8 Method, and cell apoptosis was detected by terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling. To observe further the effects of AFP-Wtp53-pPRIME-miR30-shRNA-IGF1R therapy in animals, models of BALB-C nude mice bearing subcutaneous human hepatocellular carcinoma were established. The influence of the growth of subcutaneously transplanted tumor, expression of Wtp53 protein, apoptosis, and microvessel formation on the overall level of AFP-Wtp53 pPRIME-miR30-shRNA-IGF1R were also evaluated. Results Recombinant lentivirus was successfully constructed, and its functional plaque-forming unit titer was determined as 4.58×10 9 plaque-forming units/ml. A positive strand was detected by polymerase chain reaction and Western blotting. Lentiviral construction worked effectively in AFP-positive liver cancer cells. In vitro and in vivo experiments showed that the recombinant lentivirus was more efficacious in inhibiting the proliferation of Hep3B cells. Conclusions The Wtp53-pPRIME-miR30-shRNA gene can be subjected to targeted transduction into liver cancer cells under the mediation of anti-alpha fetoprotein scFv-directed lentivirus. The Wtp53-pPRIME-miR30-shRNA system has targeting ability and lethal effects on liver cancer cells. |
| Databáze: | OpenAIRE |
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