Related Experiment Video
Updated: Aug 30, 2026
Orthotopic Transplantation of Syngeneic Lung Adenocarcinoma Cells to Study PD-L1 Expression
Published on: January 19, 2019
[Effect of HLCDG1 gene transfection on growth of lung carcinoma cells]
Fei-Yan Zou1, Hai-Long Xie, Zhu-Chu Chen
1Laboratory of Tumor Cell Biology, Cancer Research Institute, Xiangya School of Medicine, Central South University, Changsha, Hunan, PR China.
Background & Objective:
HLCDG1, which locates in chromosome 5q33 (between D5S436 and D5S470),is a novel gene that our laboratory has cloned recently. The expression of HLCDG1 gene was significantly down-regulated or deleted in the primary lung carcinoma. This study was designed to observe if HLCDG1 has the potential to suppress growth of lung carcinoma cells.
Methods:
The recombinant plasmid, pcDNA3.1(+)/HLCDG1, was constructed and subsequently transfected into A549 cells through liposome transfection. The A549 cells stably expressing HLCDG1 gene were established by G418 selection. RT-PCR was used to demonstrate the expression of HLCDG1 gene. Furthermore, the cell proliferation assay, the soft agar assay, and the tumorigenesis assay were used to analyze the malignant phenotype of the HLCDG1-transfected cells.
Results:
The HLCDG1-transfected cells exhibited the expression of HLCDG1 mRNA by RT-PCR. The population double time (PDT) of HLCDG1-transfected group, vector-transfected group, and nontransfected group were 70.0 hours, 43.3 hours, and 39.5 hours, respectively; the difference between HLCDG1-transfected group and the other two groups was significant (P< 0.05). The colony formation rates of HLCDG1-transfected group, the vector-transfected group, and nontransfected group were 8.5%, 29.0%, and 35.0%, respectively. The rate of HLCDG1-transfected cells was markedly lower than those of the other two groups (P< 0.05). Moreover, these clones were injected into athymic nude mice. After 43 days, they were killed, and their tumors were isolated. These tumors weighed 0.120g, 0.612g, and 0.924g, respectively.
Conclusion:
The expression of HLCDG1 in A549 cells may have the potential to suppress tumor cell growth and the tumorigenesis of A549 cells transplanted in nude mice. These results suggested that HLCDG1 gene might be a good candidate of tumor suppressor gene correlated with lung carcinoma.
Insights
The novel HLCDG1 gene, downregulated in lung cancer, significantly suppressed lung cancer cell growth and tumorigenesis in A549 cells. This suggests HLCDG1 may function as a tumor suppressor gene in lung carcinoma.
Area of Science:
- Genetics
- Molecular Biology
- Oncology
Context:
- The HLCDG1 gene, located on chromosome 5q33, is newly identified.
- Its expression is notably reduced or absent in primary lung carcinomas.
Purpose:
- To investigate the potential of HLCDG1 to inhibit lung carcinoma cell proliferation.
- To evaluate the role of HLCDG1 as a tumor suppressor in lung cancer models.
Summary:
- HLCDG1 was successfully expressed in A549 lung cancer cells via pcDNA3.1(+)/HLCDG1 plasmid transfection.
- RT-PCR confirmed HLCDG1 mRNA expression.
- Functional assays demonstrated that HLCDG1 significantly reduced cell proliferation (population doubling time), colony formation, and tumor growth in vivo compared to control groups.
Impact:
- HLCDG1 expression inhibits lung cancer cell growth and tumorigenesis.
- HLCDG1 shows promise as a potential tumor suppressor gene for lung carcinoma therapy.
