Publications (3)0 Total impact
Zhongguo fei ai za zhi = Chinese journal of lung cancer 04/2012; 15(4):228-33.
Article: [Lentivirus-mediated stable silencing of nm23-H1 gene in lung cancer cells and the influence on biological behavior].[show abstract] [hide abstract]
ABSTRACT: The nm23-H1 gene is an important tumor metastatic suppressor gene. Our previous study showed that the downregulation of nm23-H1 gene expression using small interfering RNA (siRNA) in NL9980 lung cancer cells greatly enhanced their invasiveness. To further explore the molecular mechanisms after nm23-H1 gene knockdown, we established transgene NL9980 and A549 lung cancer cell lines with stable nm23-H1 gene silencing through the lentivirus-mediated short hairpin RNA (shRNA) method. The human large cell lung cancer NL9980 and human lung adenocarcinoma A549 cells were transfected with shRNA lentiviral particles specific for the nm23-H1 gene, and were then selected through puromycin. Puromycin-resistant clones were generated and screened using reverse transcription polymerase chain reaction (RT-PCR), quantitative real-time polymerase chain reaction (qPCR), and Western blot analysis. shRNA rescue experiments were performed to restore the nm23-H1 gene expression in the shRNA-expressing cells. Invasiveness was determined through a Boyden chamber assay. The puromycin-resistant clones (NL9980-99 and A549-99) showed very low levels of nm23-H1 mRNA and protein expression under RT-PCR, qPCR, and Western blot analysis. Meanwhile, the shRNA rescue experiment restored the nm23-H1 expression in the NL9980-99 and A549-99 cells detected by Western blot. Downregulation of nm23-H1 gene expression enhanced the invasiveness of the NL9980-99 and A549-99 cells compared with the controls. The lung cancer cell lines NL9980-99 and A549-99 with stable nm23-H1 gene silencing were successfully established and their invasiveness was greatly increased after nm23-H1 gene knockdown.Zhongguo fei ai za zhi = Chinese journal of lung cancer 03/2012; 15(3):139-45.
Article: [Screening of metastasis-related microRNAs in the large-cell lung cancer cell lines with different metastastic potentials].[show abstract] [hide abstract]
ABSTRACT: MicroRNAs (miRNAs) regulate a wide range of cancer-associated processes, including cell division, proliferation, cell cycle, apoptosis, angiogenesis, invasion, and metastasis. A microarray was performed to analyze metastasis-related miRNAs with different metastastic potentials and to further elucidate their mechanism in the large-cell lung cancer cell lines. L9981 and NL9980 cells were harvested, and total RNA was extracted for CY3. RNA hybridization was then performed on the chip with marked miRNAs. MiRNAs with significantly different expression were selected through statistical analysis. A real-time polymerase chain reaction (PCR) was employed to validate the results of the microarray, and target genes were predicted using bioinformatics. Expressions of 22 miRNAs were significantly different in the L9981/NL9980 cell lines. Compared with the NL9980, 13 miRNAs were upregulated in the L9981 cell lines, whereas 9 miRNAs were downregulated. The result of miR-125a-3p expression based on real-time PCR was consistent with the microarray. Insulin-like growth factor 2 might be a target gene of miR-125a-3p. The metastatic miRNA profile in large-cell lung cancer was successfully screened out.Zhongguo fei ai za zhi = Chinese journal of lung cancer 11/2011; 14(11):835-40.