Related Experiment Video
Updated: Apr 22, 2026
Multidimensional Coculture System to Model Lung Squamous Carcinoma Progression
Published on: March 17, 2020
Experimental study on inhibition effects of the XAF1 gene against lung cancer cell proliferation
Wen-Tao Yang1, Dong-Lai Chen, Fu-Quan Zhang
1Department of Cardiothoracic Surgery, Second Affiliated Hospital, Soochow University, Suzhou, China
Objective:
To investigate the effect of high expression of XAF1 in vivo or in vitro on lung cancer cell growth and apoptosis.
Methods:
1. The A549 human lung cancer cell line was transfected with Ad5/F35 - XAF1, or Ad5/ F35 - Null at the same multiplicity of infection (MOI); (hereinafter referred to as transient transfected cell strain); XAF1 gene mRNA and protein expression was detected by reverse transcription polymerase chain reaction (RT- PCR) and Western blotting respectively. 2. Methyl thiazolyl tetrazolium (MTT) and annexin V-FITC/PI double staining were used to detect cell proliferation and apoptosis before and after infection of Ad5/F35 - XAF1 with Western blotting for apoptosis related proteins, caspase 3, caspase - 8 and PARP. 3. After the XAF1 gene was transfected into lung cancer A549 cells by lentiviral vectors, and selected by screening with Blasticidin, reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were applied to detect mRNA and protein expression, to establish a line with a stable high expression of XAF1 (hereinafter referred to as stable expression cell strain). Twenty nude mice were randomly divided into groups A and B, 10 in each group: A549/ XAF1 stable expression cell strain was subcutaneously injected in group A, and A549/Ctrl stable cell line stable expression cell strain in group B (control group), to observe transplanted tumor growth in nude mice.
Results:
The mRNA and protein expression of XAF1 in A549 cells transfected by Ad5/F35 - XAF1 was significantly higher than in the control group. XAF1 mediated by adenovirus vector demonstrated a dose dependent inhibition of lung cancer cell proliferation and induction of apoptosis. This was accompanied by cleavage of caspase -3, -8, -9 and PARP, suggesting activation of intrinsic or extrinsic apoptotic pathways. A cell strain of lung cancer highly expressing XAF1 was established, and this demonstrated delayed tumor growth after transplantation in vivo.
Conclusion:
Adenovirus mediated XAF1 gene expression could inhibit proliferation and induce apoptosis in lung cancer cells in vitro; highly stable expression of XAF1 could also significantly inhibit the growth of transplanted tumors in nude mouse, with no obvious adverse reactions observed. Therefore, the XAF1 gene could become a new target for lung cancer treatment.
Insights
High expression of the X-linked inhibitor of apoptosis-associated factor 1 (XAF1) gene inhibits lung cancer cell growth and induces apoptosis. This finding suggests XAF1 as a potential new therapeutic target for lung cancer treatment.
Area of Science:
- Oncology
- Molecular Biology
- Gene Therapy
Background:
- Lung cancer remains a leading cause of cancer-related mortality worldwide.
- Identifying novel therapeutic targets is crucial for improving patient outcomes.
- The role of X-linked inhibitor of apoptosis-associated factor 1 (XAF1) in lung cancer pathogenesis requires further investigation.
Purpose of the Study:
- To investigate the in vitro and in vivo effects of high XAF1 expression on lung cancer cell growth and apoptosis.
- To evaluate the potential of XAF1 as a therapeutic target for lung cancer.
Main Methods:
- Adenovirus-mediated transfection of A549 lung cancer cells with XAF1.
- Assessment of XAF1 mRNA and protein expression using RT-PCR and Western blotting.
- Evaluation of cell proliferation and apoptosis via MTT assays and Annexin V-FITC/PI staining.
- Establishment of a stable XAF1-expressing cell line using lentiviral vectors.
- In vivo studies involving subcutaneous injection of XAF1-expressing cells into nude mice to monitor tumor growth.
Main Results:
- Adenovirus-mediated XAF1 expression significantly inhibited lung cancer cell proliferation and induced apoptosis in vitro.
- XAF1 overexpression led to the activation of caspase-3, -8, -9, and PARP, indicating the involvement of intrinsic and extrinsic apoptotic pathways.
- Stable high expression of XAF1 in lung cancer cells resulted in delayed tumor growth in vivo.
- No significant adverse reactions were observed in the in vivo model.
Conclusions:
- Adenovirus-mediated XAF1 gene delivery effectively inhibits lung cancer cell proliferation and promotes apoptosis.
- Stable high expression of XAF1 significantly suppresses transplanted tumor growth in vivo.
- XAF1 represents a promising novel therapeutic target for lung cancer treatment.
