[Study on combined gene therapy for malignant gliomas transfected with antisense hTERT/PTEN in vitro and in vivo]
Yong-ping You1, Zhen Fu, Peng Zhao
1Department of Neurosurgery, First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210029, P.R. China.
Objective:
To study inhibitory efficacy of combined gene therapy for malignant gliomas transfected with antisense human telomerase reverse transcriptase (hTERT)/PTEN in vitro and in vivo.
Methods:
To construct two adenovirus recons which contained antisense hTERT and wild-type PTEN respectively with high performance homologous recombination system in bacteria. The two adenovirus recons were transfected into U251 human malignant glioma cells combinedly or respectively in vitro and in vivo. U251 cell proliferation in vitro was determined by MTT assay and flow cytometry, tumor growth in vivo was measured by the volume of glioma in nude mice. Telomerase activity was detected by telomeric repeat amplification protocol (TRAP) assay. Expression of hTERT and PTEN protein was detected by Western blotting methods.
Results:
After transfection in vitro, the growth of U251 cells was inhibited significantly. The inhibitory effect was time-dependent. The strongest inhibition was observed in combined transfection group, at the 6th day, the survival rate was 37.6%, telomerase activity (only 28.8TPG) was inhibited significantly, hTERT protein expression was inhibited significantly too, which was 0.2106, but PTEN protein expression was increased significantly, which was 0.9630. In vivo, the growth of tumors was also effectively inhibited.
Conclusion:
Growth of malignant glioma cells is effectively inhibited after transfection with combined antisense hTERT and PTEN in vitro and in vivo.
Insights
Combined gene therapy using antisense human telomerase reverse transcriptase (hTERT) and PTEN effectively inhibits malignant glioma cell growth in vitro and in vivo. This approach offers a promising strategy for treating aggressive brain tumors.
Area of Science:
- Oncology
- Molecular Biology
- Gene Therapy
Context:
- Malignant gliomas are aggressive brain tumors with limited treatment options.
- Human telomerase reverse transcriptase (hTERT) is crucial for tumor cell proliferation.
- PTEN is a tumor suppressor gene often inactivated in cancers.
Purpose:
- To evaluate the inhibitory efficacy of combined gene therapy using antisense hTERT and PTEN.
- To assess the in vitro and in vivo effects on malignant glioma cells.
Summary:
- Adenoviruses encoding antisense hTERT and wild-type PTEN were constructed and transfected into U251 glioma cells.
- Combined transfection significantly inhibited cell proliferation, reduced telomerase activity, and decreased hTERT expression while increasing PTEN expression.
- Tumor growth was also effectively inhibited in vivo.
Impact:
- Demonstrates the potential of combined gene therapy targeting hTERT and PTEN as a treatment strategy for malignant gliomas.
- Provides a foundation for further preclinical and clinical investigations.
- Highlights the synergistic effect of inhibiting hTERT and restoring PTEN function.
