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Transcription profiling by array of stem cells isolated from non-small cell lung cancer cell lines (NCI-H2170 and A549) and normal lung epithelial cell line (PHBEC)

ABSTRACT: This study was designed to understand the transcriptomic composition and the biological functions of cancer stem cells isolated from non-small cell lung cancer line (NSCLC) Putative lung cancer stem cells were isolated from cancer cell lines based on expression of known stem cell surface markers: CD166, CD44 and EpCAM using the Fluorescence Activated Cell Sorter (FACS). Affymetrix microarray were performed on cancer stem cells isolated from normal lung epithelial cells and lung cancer cell lines (A549 and NCI-H2170) using GeneChip Human Gene 1.0 ST array. The normal putative stem cells isolated from normal primary human bronchial/trachial epithelial cell line (PHBEC) was serve as control. Putative cancer stem cells isolated from A549 and NCI-H2170 cell lines are the treatment group. Each sample was performed in triplicate and total number of samples are five (n=5)

ORGANISM(S): Homo sapiens

SUBMITTER: Yahaya Badrul

PROVIDER: S-ECPF-GEOD-50627 | biostudies-other |

REPOSITORIES: biostudies-other

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Project description:In both males and females, lung cancer is one of the most lethal cancers worldwide and accounts for >30% of cancer-related deaths. Despite advances in biomarker analysis and tumor characterization, there remains a need to find suitable biomarker antigen targets for treatment in late-stage lung cancer. Previous research on the salvage pathway enzyme TK1 shows a unique relationship with cancer patients as serum levels are raised according to cancer grade. To expand this analysis, the other salvage pathway enzymes were evaluated for possible upregulation within lung cancer. Adenine phosphoribosyltransferase, deoxycytidine kinase, and hypoxanthine guanine phosphoribosyltransferase (HPRT) were assessed for their presentation on two non-small-cell lung cancer cell lines NCI-H460 and A549. In the present study, we show that deoxycytidine kinase and adenine phosphoribosyltransferase have no significant relationship with the membrane of NCI-H460 cells. However, we found significant localization of HPRT to the membrane of NCI-H460 and A549 cells. When treated with anti-HPRT antibodies, the average fluorescence of the cell population increased by 24.3% and 12.9% in NCI-H460 and A549 cells, respectively, in comparison with controls. To ensure that expression was not attributed to cytoplasmic HPRT, confocal microscopy was performed to visualize HPRT binding on the plasma membrane. After staining NCI-H460 cells treated with both fluorescent antibodies and a membrane-specific dye, we observed direct overlap between HPRT and the membrane of the cancer cells. Additionally, gold-conjugated antibodies were used to label and quantify the amount of HPRT on the cell surface using scanning electron microscopy and energy-dispersive analysis X-ray. Further confirming HPRT presence, the gold weight percentage of the sample increased significantly when NCI-H460 cells were exposed to HPRT antibody (P=0.012) in comparison with isotype controls. Our results show that HPRT is localized on the surface of these non-small-cell lung cancer cell lines.

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Transcription profiling by array of stem cells isolated from non-small cell lung cancer cell lines (NCI-H2170 and A549) and normal lung epithelial cell line (PHBEC)

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