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Non-small-cell lung cancer cell lines A549 and NCI-H460 express hypoxanthine guanine phosphoribosyltransferase on the plasma membrane

View Article: PubMed Central - PubMed

ABSTRACT

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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Levels of HPRT expression compared between A549 and H460 cells. Notes: While both A549 and H460 cells show a statistically significant increase in the surface expression of HPRT, H460 cells had a significantly higher expression (P<0.0001). H460 cells are a faster growing cell line, with a growth rate almost double that of A549 cells. As a result, HPRT expression on the surface of non-small-cell lung cancer cells may directly correspond to cell proliferation. ****P<0.0001.Abbreviation: HPRT, hypoxanthine guanine phosphoribosyltransferase.
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f3-ott-10-1921: Levels of HPRT expression compared between A549 and H460 cells. Notes: While both A549 and H460 cells show a statistically significant increase in the surface expression of HPRT, H460 cells had a significantly higher expression (P<0.0001). H460 cells are a faster growing cell line, with a growth rate almost double that of A549 cells. As a result, HPRT expression on the surface of non-small-cell lung cancer cells may directly correspond to cell proliferation. ****P<0.0001.Abbreviation: HPRT, hypoxanthine guanine phosphoribosyltransferase.

Mentions: When treated with anti-HPRT fluorescent antibodies, both A549 and H460 cancer cells had an increase in the fluorescent population (Figures 1C and 2). A 28% shift in the population is observed in H460 cells (Figure 1C), while a 12% shift is observed in A549 cells (Figure 3). Statistical analysis comparing anti-HPRT-treated cells with isotype IgG controls showed a statistically significant difference in H460 and A549 cells (Figures 1C and 2C). Thus, these data show a significant association between HPRT and the surface of non-small-cell lung cancer cells. This analysis also revealed a significantly higher HPRT surface expression in H460 cells when compared to A549 (Figure 3).


Non-small-cell lung cancer cell lines A549 and NCI-H460 express hypoxanthine guanine phosphoribosyltransferase on the plasma membrane
Levels of HPRT expression compared between A549 and H460 cells. Notes: While both A549 and H460 cells show a statistically significant increase in the surface expression of HPRT, H460 cells had a significantly higher expression (P<0.0001). H460 cells are a faster growing cell line, with a growth rate almost double that of A549 cells. As a result, HPRT expression on the surface of non-small-cell lung cancer cells may directly correspond to cell proliferation. ****P<0.0001.Abbreviation: HPRT, hypoxanthine guanine phosphoribosyltransferase.
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Related In: Results  -  Collection

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f3-ott-10-1921: Levels of HPRT expression compared between A549 and H460 cells. Notes: While both A549 and H460 cells show a statistically significant increase in the surface expression of HPRT, H460 cells had a significantly higher expression (P<0.0001). H460 cells are a faster growing cell line, with a growth rate almost double that of A549 cells. As a result, HPRT expression on the surface of non-small-cell lung cancer cells may directly correspond to cell proliferation. ****P<0.0001.Abbreviation: HPRT, hypoxanthine guanine phosphoribosyltransferase.
Mentions: When treated with anti-HPRT fluorescent antibodies, both A549 and H460 cancer cells had an increase in the fluorescent population (Figures 1C and 2). A 28% shift in the population is observed in H460 cells (Figure 1C), while a 12% shift is observed in A549 cells (Figure 3). Statistical analysis comparing anti-HPRT-treated cells with isotype IgG controls showed a statistically significant difference in H460 and A549 cells (Figures 1C and 2C). Thus, these data show a significant association between HPRT and the surface of non-small-cell lung cancer cells. This analysis also revealed a significantly higher HPRT surface expression in H460 cells when compared to A549 (Figure 3).

View Article: PubMed Central - PubMed

ABSTRACT

In both males and females, lung cancer is one of the most lethal cancers worldwide and accounts for &gt;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.

No MeSH data available.


Related in: MedlinePlus