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[Effect of nm23-H1 on reversing malignant phenotype on human lung cancer cell line L9981]
Guowei Che1, Qinghua Zhou, Yanping Wang
1The Key Laboratory of Lung Cancer Molecular Biology, West China Hospital, Sichuan University, Chengdu 610041, China.
Abstract:
nm23-H1 is a proven tumor metastasis suppressive gene, tumor metastasis phenotype could be reversed by transfected nm23-H1 cDNA. This study was conducted to transfect nm23-H1 cDNA into L9981 cells and to explore the function of nm23-H1 in reversing the malignant phenotype of L9981 cells. The plasmid of pLXSN-nm23-H1-EGFP was constructed by gene clone technique, and the transfected nm23-H1 cDNA cell lines of L9981-nm23-H1 was established. The protein expression of nm23-H1 was detected by Western blot. The biologic features of L9981-nm23-H1 cells were studied in vitro and in vivo. The results showed that the fusion protein of nm23-H1-EGFP was stable, continuous and expressed with high efficiency in L9981-nm23-H1 cells. The cell proliferation, colon formation and invasive ability are significantly lowered in L9981 cells transfected nm23-H1 cDNA (P < 0.01); the tumorgenesis and the lung metastasis incidence was lower in tranfected nm23-H1 cells than in L9981 and L9981-Plxsn in nude mice (P < 0.01); the rate for inhibiting tumorgenesis of nm23 -H1 was 82.56%. These data suggest that the malignant phenotype could be reversed by wild nm23-H1 gene in L9981 cells.
Insights
nm23-H1 gene transfection significantly suppressed tumor metastasis and reversed malignant phenotypes in L9981 cells. This study demonstrates nm23-H1
Area of Science:
- Molecular Biology
- Oncology
- Gene Therapy
Background:
- nm23-H1 is a known tumor metastasis suppressive gene.
- Reversing tumor metastasis phenotype is achievable through nm23-H1 cDNA transfection.
- L9981 cells exhibit a malignant phenotype that can be targeted.
Purpose of the Study:
- To investigate the function of nm23-H1 in reversing the malignant phenotype of L9981 cells.
- To establish L9981 cell lines stably expressing nm23-H1.
- To evaluate the impact of nm23-H1 on L9981 cell behavior in vitro and in vivo.
Main Methods:
- Construction of pLXSN-nm23-H1-EGFP plasmid using gene cloning.
- Establishment of L9981-nm23-H1 cell lines via transfection.
- Detection of nm23-H1 protein expression using Western blot.
- Assessment of biological features in vitro and in vivo.
Main Results:
- nm23-H1-EGFP fusion protein showed stable, high-efficiency expression in transfected cells.
- Transfection significantly reduced L9981 cell proliferation, colony formation, and invasion (P < 0.01).
- Tumorigenesis and lung metastasis incidence were significantly lower in nm23-H1 transfected cells in nude mice (P < 0.01).
- nm23-H1 inhibited tumorigenesis by 82.56%.
Conclusions:
- Wild-type nm23-H1 gene can effectively reverse the malignant phenotype of L9981 cells.
- nm23-H1 demonstrates potent anti-metastatic and anti-tumorigenic properties.
- Gene therapy targeting nm23-H1 holds potential for cancer treatment.
