Limits...
Changes in tumor-antigen expression profile as human small-cell lung cancers progress.

Ge LS, Hoa NT, Lambrecht N, Dacosta-Iyer M, Ouyang Y, Abolhoda A, Jadus MR - Cancer Biol Med (2015)

Bottom Line: Our group has previously observed that in patients with small-cell lung cancers (SCLCs), the expression of a tumor antigen, glioma big potassium (gBK) ion channel, is higher at the time of death than when the cancer is first treated by surgical resection.When HTB119 cells were incubated with doxorubicin, gBK was strongly induced, as confirmed by intracellular flow cytometry with a gBK-specific antibody.Our findings suggested that more immunological targets became available as the tumor responded to chemotherapy and proceeded toward its terminal stages.

View Article: PubMed Central - PubMed

Affiliation: 1 Research Service, 2 Pathology and Laboratory Medicine Service, VA Long Beach Healthcare System, Long Beach, CA 90822, USA ; 3 Pathology and Laboratory Medicine, University of California, Irvine, CA 92697, USA ; 4 Surgical Health Care Group, Veterans Affairs Medical Center, Long Beach, CA 90822, USA ; 5 Chao Family Comprehensive Cancer Center, UC, Irvine School of Medicine, University of California, Irvine, Orange, CA 92868, USA.

ABSTRACT

Objective: Our group has previously observed that in patients with small-cell lung cancers (SCLCs), the expression of a tumor antigen, glioma big potassium (gBK) ion channel, is higher at the time of death than when the cancer is first treated by surgical resection. This study aimed to determine whether this dichotomy was common in other potential lung tumor antigens by examining the same patient samples using our more extensive profile analysis of tumor-antigen precursor protein (TAPP). We then tested the hypothesis that therapeutic intervention may inadvertently cause this increased gBK production.

Methods: SCLC samples (eight surgical resections and three autopsy samples) and three control lungs were examined by quantitative real-time polymerase chain reaction for 42 potential TAPPs that represent potential T-cell-mediated immunological targets.

Results: Twenty-two TAPP mRNAs displayed the same profile as gBK, i.e., more mRNAs were expressed at autopsy than in their surgical counterparts. B-cyclin and mouse double minute 2, human homolog of P53-binding protein were elevated in both autopsy and surgical specimens above the normal-lung controls. When HTB119 cells were incubated with doxorubicin, gBK was strongly induced, as confirmed by intracellular flow cytometry with a gBK-specific antibody.

Conclusion: Our findings suggested that more immunological targets became available as the tumor responded to chemotherapy and proceeded toward its terminal stages.

No MeSH data available.


Related in: MedlinePlus

gBK protein is induced with HTB119 and H1436 by doxorubicin. The exact same chemotherapeutic conditions from (Figure 5) were repeated using both HTB119 and H1436 SCLC cells. After 2 weeks of incubation, the cells were fixed, permeabilized and stained for gBK using the goat anti-human gBK antibody. After washing, they were stained with a mouse anti-goat IgG-FITC antibody. Subsequently these washed cells were analyzed by flow cytometry. Ten thousand cells were examined. The shaded area represents the fluorescence of the DMSO-treated controls stained for gBK, while the open area is the profile of the drug treated cells stained for gBK. CIS is Cis-platinum treated cells, CYC is cyclophosphamide treated cells, DOX is doxorubicin treated cells and ETO is Etoposide treated cell.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4493377&req=5

f6: gBK protein is induced with HTB119 and H1436 by doxorubicin. The exact same chemotherapeutic conditions from (Figure 5) were repeated using both HTB119 and H1436 SCLC cells. After 2 weeks of incubation, the cells were fixed, permeabilized and stained for gBK using the goat anti-human gBK antibody. After washing, they were stained with a mouse anti-goat IgG-FITC antibody. Subsequently these washed cells were analyzed by flow cytometry. Ten thousand cells were examined. The shaded area represents the fluorescence of the DMSO-treated controls stained for gBK, while the open area is the profile of the drug treated cells stained for gBK. CIS is Cis-platinum treated cells, CYC is cyclophosphamide treated cells, DOX is doxorubicin treated cells and ETO is Etoposide treated cell.

Mentions: We showed that elevated gBK mRNA expression was translated into protein by repeating the previous experiment and performing intracellular flow cytometry with the polyclonal antibody to gBK. Figure 6 shows that doxorubicin induced gBK expression in both HTB119 (top row) and another SCLC cell line, H1436 (bottom row). Etoposide significantly induced gBK protein expression in H1436 cells.


Changes in tumor-antigen expression profile as human small-cell lung cancers progress.

Ge LS, Hoa NT, Lambrecht N, Dacosta-Iyer M, Ouyang Y, Abolhoda A, Jadus MR - Cancer Biol Med (2015)

gBK protein is induced with HTB119 and H1436 by doxorubicin. The exact same chemotherapeutic conditions from (Figure 5) were repeated using both HTB119 and H1436 SCLC cells. After 2 weeks of incubation, the cells were fixed, permeabilized and stained for gBK using the goat anti-human gBK antibody. After washing, they were stained with a mouse anti-goat IgG-FITC antibody. Subsequently these washed cells were analyzed by flow cytometry. Ten thousand cells were examined. The shaded area represents the fluorescence of the DMSO-treated controls stained for gBK, while the open area is the profile of the drug treated cells stained for gBK. CIS is Cis-platinum treated cells, CYC is cyclophosphamide treated cells, DOX is doxorubicin treated cells and ETO is Etoposide treated cell.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493377&req=5

f6: gBK protein is induced with HTB119 and H1436 by doxorubicin. The exact same chemotherapeutic conditions from (Figure 5) were repeated using both HTB119 and H1436 SCLC cells. After 2 weeks of incubation, the cells were fixed, permeabilized and stained for gBK using the goat anti-human gBK antibody. After washing, they were stained with a mouse anti-goat IgG-FITC antibody. Subsequently these washed cells were analyzed by flow cytometry. Ten thousand cells were examined. The shaded area represents the fluorescence of the DMSO-treated controls stained for gBK, while the open area is the profile of the drug treated cells stained for gBK. CIS is Cis-platinum treated cells, CYC is cyclophosphamide treated cells, DOX is doxorubicin treated cells and ETO is Etoposide treated cell.
Mentions: We showed that elevated gBK mRNA expression was translated into protein by repeating the previous experiment and performing intracellular flow cytometry with the polyclonal antibody to gBK. Figure 6 shows that doxorubicin induced gBK expression in both HTB119 (top row) and another SCLC cell line, H1436 (bottom row). Etoposide significantly induced gBK protein expression in H1436 cells.

Bottom Line: Our group has previously observed that in patients with small-cell lung cancers (SCLCs), the expression of a tumor antigen, glioma big potassium (gBK) ion channel, is higher at the time of death than when the cancer is first treated by surgical resection.When HTB119 cells were incubated with doxorubicin, gBK was strongly induced, as confirmed by intracellular flow cytometry with a gBK-specific antibody.Our findings suggested that more immunological targets became available as the tumor responded to chemotherapy and proceeded toward its terminal stages.

View Article: PubMed Central - PubMed

Affiliation: 1 Research Service, 2 Pathology and Laboratory Medicine Service, VA Long Beach Healthcare System, Long Beach, CA 90822, USA ; 3 Pathology and Laboratory Medicine, University of California, Irvine, CA 92697, USA ; 4 Surgical Health Care Group, Veterans Affairs Medical Center, Long Beach, CA 90822, USA ; 5 Chao Family Comprehensive Cancer Center, UC, Irvine School of Medicine, University of California, Irvine, Orange, CA 92868, USA.

ABSTRACT

Objective: Our group has previously observed that in patients with small-cell lung cancers (SCLCs), the expression of a tumor antigen, glioma big potassium (gBK) ion channel, is higher at the time of death than when the cancer is first treated by surgical resection. This study aimed to determine whether this dichotomy was common in other potential lung tumor antigens by examining the same patient samples using our more extensive profile analysis of tumor-antigen precursor protein (TAPP). We then tested the hypothesis that therapeutic intervention may inadvertently cause this increased gBK production.

Methods: SCLC samples (eight surgical resections and three autopsy samples) and three control lungs were examined by quantitative real-time polymerase chain reaction for 42 potential TAPPs that represent potential T-cell-mediated immunological targets.

Results: Twenty-two TAPP mRNAs displayed the same profile as gBK, i.e., more mRNAs were expressed at autopsy than in their surgical counterparts. B-cyclin and mouse double minute 2, human homolog of P53-binding protein were elevated in both autopsy and surgical specimens above the normal-lung controls. When HTB119 cells were incubated with doxorubicin, gBK was strongly induced, as confirmed by intracellular flow cytometry with a gBK-specific antibody.

Conclusion: Our findings suggested that more immunological targets became available as the tumor responded to chemotherapy and proceeded toward its terminal stages.

No MeSH data available.


Related in: MedlinePlus