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An Oncogenic Hepatocyte-Induced Orthotopic Mouse Model of Hepatocellular Cancer Arising in the Setting of Hepatic Inflammation and Fibrosis
Published on: September 12, 2019
Multi-target lentivirus specific to hepatocellular carcinoma: in vitro and in vivo studies
Ye-Wei Zhang1, Jian Niu, Xiang Lu
1Department of Hepatobiliary & Pancreatic Surgery, Affiliated Jiangsu Cancer Hospital of Nanjing Medical University, Nanjing 210009, China. zhangyewei@njmu.edu.cn
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.
Insights
Targeted delivery of Wtp53-pPRIME-miR30-shRNA using lentivirus effectively inhibited liver cancer cell growth. This novel gene therapy shows promise for treating hepatocellular carcinoma by targeting AFP-positive cells.
Area of Science:
- Oncology
- Gene Therapy
- Molecular Biology
Background:
- Hepatocellular carcinoma (HCC) remains a significant global health challenge.
- Targeted gene delivery offers a promising strategy for HCC treatment.
- Developing effective therapeutic vectors for HCC is crucial.
Purpose of the Study:
- To investigate the targeted transduction of the Wtp53-pPRIME-miR30-shRNA gene into liver cancer cells.
- To evaluate the inhibitory effect of this gene therapy system on liver cancer.
- To assess the efficacy of anti-alpha fetoprotein scFv-directed lentivirus mediated gene delivery.
Main Methods:
- Construction and characterization of recombinant lentivirus carrying Wtp53-pPRIME-miR30-shRNA.
- In vitro assessment of gene transduction, cell proliferation (Cell-Counting Kit-8), and apoptosis (TUNEL assay).
- In vivo evaluation in BALB-C nude mice models of human hepatocellular carcinoma.
Main Results:
- Successful construction and functional titration of the recombinant lentivirus.
- Demonstrated effective transduction and transcription of the Wtp53-pPRIME-miR30-shRNA-IGF1R gene in AFP-positive liver cancer cells.
- Significant inhibition of Hep3B cell proliferation in vitro and tumor growth in vivo.
Conclusions:
- The Wtp53-pPRIME-miR30-shRNA system, delivered via anti-AFP scFv lentivirus, exhibits targeted transduction into liver cancer cells.
- This therapeutic system demonstrates significant targeting ability and potent inhibitory effects on liver cancer cells.
- The findings support the potential of this targeted gene therapy for hepatocellular carcinoma treatment.
