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[Plasmid-mediated expression of kallistatin and its biological activity in lung cancer related cells]
Ning-Qing Wang1, Jin Zou, Yong Diao
1School of Biomedical Science, Institutes of Molecular Medicine, Huaqiao University, Quanzhou 362021, China.
Abstract:
This study is to investigate whether naked plasmid DNA can effectively transfect lung cancer related cells and express human kallistatin, an endogenous protein that inhibits angiogenesis and tumor growth, and to explore the biological activity of the low-level expressed kallistatin to lung cancer in vitro and in vivo. The plasmids were delivered with Lipofectamine 2000 to transfect various lung cancer related cells. Kal expression was determined by ELISA. The biological effects of Kal expression on proliferation, migration and apoptosis rate of the cells were examined. In subcutaneous NCI-H446 xenograft model, pKal was injected directly into tumors, the changes of CD34, Ki-67 and E-cadherin expression were detected with immunohistochemical assay, the tumor apoptosis was analyzed with TUNEL assay. Both the endothelial cell and lung cancer cells could express kallistatin after plasmid transfection. The proliferation and migration of human umbilical vein endothelial cells were inhibited, but the apoptosis rate was not affected. The proliferation rates of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited, and their apoptosis rates were enhanced, but different cells behaved differently. In subcutaneous NCI-H446 xenograft model, intratumor injection of pKal inhibited the growth of lung cancer by reducing angiogenesis and proliferation of tumor cells. In conclusion, this study demonstrated the efficacy of plasmid-mediated expression of kallistatin to lung cancer related cells, thus providing a basis for their clinical application in the treatment of lung cancer.
Insights
Naked plasmid DNA effectively delivered human kallistatin (Kal) to lung cancer cells, inhibiting tumor growth and angiogenesis. This gene therapy approach shows promise for lung cancer treatment.
Area of Science:
- Molecular Biology
- Cancer Research
- Gene Therapy
Context:
- Lung cancer remains a leading cause of cancer-related mortality worldwide.
- Novel therapeutic strategies are crucial for improving patient outcomes.
- Gene therapy offers a potential avenue for targeted cancer treatment.
Purpose:
- To investigate the efficacy of naked plasmid DNA for transfecting lung cancer cells and expressing human kallistatin (Kal).
- To explore the biological activity of Kal in inhibiting lung cancer proliferation, migration, and angiogenesis in vitro and in vivo.
- To assess the potential of Kal gene therapy for clinical application in lung cancer treatment.
Summary:
- Plasmid DNA with Lipofectamine 2000 successfully transfected lung cancer cells, leading to kallistatin expression.
- Kallistatin inhibited proliferation and migration of endothelial cells and lung cancer cells (NCI-H460, A549, NCI-H446), while enhancing apoptosis in cancer cells.
- In vivo studies demonstrated that intratumoral kallistatin injection reduced angiogenesis and tumor growth in a xenograft model.
Impact:
- This study validates plasmid-mediated kallistatin expression as a viable strategy against lung cancer.
- The findings provide a strong foundation for developing kallistatin-based gene therapies for lung cancer.
- Successful inhibition of tumor growth and angiogenesis highlights the therapeutic potential of kallistatin.
