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Prognostic significance of thymidylate synthase in postoperative non-small cell lung cancer patients.

Zhao HY, Ma GW, Zou BY, Li M, Lin SX, Zhao LP, Guo Y, Huang Y, Tian Y, Xie D, Zhang L - Onco Targets Ther (2014)

Bottom Line: Compared with the TS-positive patients, the overall survival (OS) was significantly lower in the TS-negative patients (hazard ratio [HR] =1.766, 95% confidence interval [CI] =1.212-2.573, P=0.003).Significant differences between TS-positive and TS-negative patients was also observed in the following stratified analyses: 1) adenocarcinoma subgroup (HR =2.079, 95% CI =1.235-3.500, P=0.006); 2) less than 60-year-old subgroup (HR =1.890, 95% CI =1.061-3.366, P=0.031); 3) stage II/III subgroup (HR =1.594, 95% CI =1.036-2.453, P=0.034); and 4) surgery plus adjuvant therapy subgroup (HR =1.976, 95% CI =1.226-3.185, P=0.005).However, the OS was not significantly correlated with OPRT or TP protein expression.

View Article: PubMed Central - PubMed

Affiliation: Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, People's Republic of China.

ABSTRACT
The aim of the present study was to investigate the clinicopathologic/prognostic significance of thymidylate synthase (TS), orotate phosphoribosyltransferase (OPRT), and thymidine phosphorylase (TP) proteins in postoperative non-small cell lung cancer (NSCLC) patients. Microarray slides from a set of 178 NSCLC patients were used for the detection of TS, OPRT, and TP expression by immunohistochemistry. The correlation between clinicopathologic factors and protein expression of three proteins was analyzed. Ninety seven carcinomas (57.4%) were TS-positive, 90 carcinomas (53.9%) were OPRT-positive, and 102 carcinomas (69.4%) were TP-positive. Compared with the TS-positive patients, the overall survival (OS) was significantly lower in the TS-negative patients (hazard ratio [HR] =1.766, 95% confidence interval [CI] =1.212-2.573, P=0.003). Significant differences between TS-positive and TS-negative patients was also observed in the following stratified analyses: 1) adenocarcinoma subgroup (HR =2.079, 95% CI =1.235-3.500, P=0.006); 2) less than 60-year-old subgroup (HR =1.890, 95% CI =1.061-3.366, P=0.031); 3) stage II/III subgroup (HR =1.594, 95% CI =1.036-2.453, P=0.034); and 4) surgery plus adjuvant therapy subgroup (HR =1.976, 95% CI =1.226-3.185, P=0.005). However, the OS was not significantly correlated with OPRT or TP protein expression. This study demonstrates that the TS level in tumor tissues may be a useful marker to predict the postoperative OS in NSCLC patients.

No MeSH data available.


Related in: MedlinePlus

Representative images of immunohistochemical staining in human NSCLC tissues (original magnification, ×200).Notes: (A) TS-positive expression; (B) TS-negative control; (C) OPRT-positive expression; (D) OPRT-negative control; (E) TP-positive expression; and (F) TP-negative control.Abbreviations: NSCLC, non-small cell lung cancer; OPRT, orotate phosphoribosyltransferase; TP, thymidine phosphorylase; TS, thymidylate synthase.
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f1-ott-7-1301: Representative images of immunohistochemical staining in human NSCLC tissues (original magnification, ×200).Notes: (A) TS-positive expression; (B) TS-negative control; (C) OPRT-positive expression; (D) OPRT-negative control; (E) TP-positive expression; and (F) TP-negative control.Abbreviations: NSCLC, non-small cell lung cancer; OPRT, orotate phosphoribosyltransferase; TP, thymidine phosphorylase; TS, thymidylate synthase.

Mentions: The primary antibodies used in this study were mouse anti-human TS monoclonal antibody (MAB4130, clone TS106, Merck Millipore, Billerica, MA, USA) (1:50 dilution in phosphate-buffered saline [PBS]); Rabbit anti-human OPRT polyclonal antibody (14830-1-AP, Proteintech Group, Chicago, IL, USA) (1:100 dilution in PBS); and mouse anti-human TP monoclonal antibody (ab3135, Abcam plc, Cambridge, UK) (1:50 dilution in PBS). Multiple 5 μm TMA sections were deparaffinized and rehydrated through graded alcohol. Prior to immunostaining, the TMA sections (for TS and OPRT staining) were treated by microwaving for 23 minutes in 10 mM citrate buffer (pH 6.0). For antigen retrieval, tissue slides were boiled in 10 mM citrate buffer (pH 6.0) in a pressure cooker for 10 minutes (for TS and OPRT) or in a microwave for 10 minutes (for TP). The sections were treated with 0.3% hydrogen peroxide for 15 minutes to quench the endogenous peroxidase activity, rinsed in 150 mM PBS (pH 7.6), then incubated with the primary antibodies at 4°C overnight. All incubation was performed in a moist chamber. Subsequently, the slides were incubated with appropriate biotinylated secondary antibodies (PV-6001 Two-Step IHC Detection Reagent, Merck Millipore, Billerica, MA, USA) at a concentration of 1:100 for 30 minutes at 37°C and then reacted with streptavidin-peroxidase conjugate for 30 minutes at 37°C and 3′-3′ diaminobenzidine (DAB; K5007 substrate buffer DAB+ Chromogen, Dako Denmark A/S, Glostrup, Denmark) as a chromogen substrate. The nucleus was counterstained using Meyer’s hematoxylin. A negative control was obtained by replacing the primary antibody with normal murine IgG. The labeling indices for TS, TP, and OPRT proteins were determined by calculating the percentage of immunoreactive cells in more than 500 cancer cells (Figure 1).


Prognostic significance of thymidylate synthase in postoperative non-small cell lung cancer patients.

Zhao HY, Ma GW, Zou BY, Li M, Lin SX, Zhao LP, Guo Y, Huang Y, Tian Y, Xie D, Zhang L - Onco Targets Ther (2014)

Representative images of immunohistochemical staining in human NSCLC tissues (original magnification, ×200).Notes: (A) TS-positive expression; (B) TS-negative control; (C) OPRT-positive expression; (D) OPRT-negative control; (E) TP-positive expression; and (F) TP-negative control.Abbreviations: NSCLC, non-small cell lung cancer; OPRT, orotate phosphoribosyltransferase; TP, thymidine phosphorylase; TS, thymidylate synthase.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4109640&req=5

f1-ott-7-1301: Representative images of immunohistochemical staining in human NSCLC tissues (original magnification, ×200).Notes: (A) TS-positive expression; (B) TS-negative control; (C) OPRT-positive expression; (D) OPRT-negative control; (E) TP-positive expression; and (F) TP-negative control.Abbreviations: NSCLC, non-small cell lung cancer; OPRT, orotate phosphoribosyltransferase; TP, thymidine phosphorylase; TS, thymidylate synthase.
Mentions: The primary antibodies used in this study were mouse anti-human TS monoclonal antibody (MAB4130, clone TS106, Merck Millipore, Billerica, MA, USA) (1:50 dilution in phosphate-buffered saline [PBS]); Rabbit anti-human OPRT polyclonal antibody (14830-1-AP, Proteintech Group, Chicago, IL, USA) (1:100 dilution in PBS); and mouse anti-human TP monoclonal antibody (ab3135, Abcam plc, Cambridge, UK) (1:50 dilution in PBS). Multiple 5 μm TMA sections were deparaffinized and rehydrated through graded alcohol. Prior to immunostaining, the TMA sections (for TS and OPRT staining) were treated by microwaving for 23 minutes in 10 mM citrate buffer (pH 6.0). For antigen retrieval, tissue slides were boiled in 10 mM citrate buffer (pH 6.0) in a pressure cooker for 10 minutes (for TS and OPRT) or in a microwave for 10 minutes (for TP). The sections were treated with 0.3% hydrogen peroxide for 15 minutes to quench the endogenous peroxidase activity, rinsed in 150 mM PBS (pH 7.6), then incubated with the primary antibodies at 4°C overnight. All incubation was performed in a moist chamber. Subsequently, the slides were incubated with appropriate biotinylated secondary antibodies (PV-6001 Two-Step IHC Detection Reagent, Merck Millipore, Billerica, MA, USA) at a concentration of 1:100 for 30 minutes at 37°C and then reacted with streptavidin-peroxidase conjugate for 30 minutes at 37°C and 3′-3′ diaminobenzidine (DAB; K5007 substrate buffer DAB+ Chromogen, Dako Denmark A/S, Glostrup, Denmark) as a chromogen substrate. The nucleus was counterstained using Meyer’s hematoxylin. A negative control was obtained by replacing the primary antibody with normal murine IgG. The labeling indices for TS, TP, and OPRT proteins were determined by calculating the percentage of immunoreactive cells in more than 500 cancer cells (Figure 1).

Bottom Line: Compared with the TS-positive patients, the overall survival (OS) was significantly lower in the TS-negative patients (hazard ratio [HR] =1.766, 95% confidence interval [CI] =1.212-2.573, P=0.003).Significant differences between TS-positive and TS-negative patients was also observed in the following stratified analyses: 1) adenocarcinoma subgroup (HR =2.079, 95% CI =1.235-3.500, P=0.006); 2) less than 60-year-old subgroup (HR =1.890, 95% CI =1.061-3.366, P=0.031); 3) stage II/III subgroup (HR =1.594, 95% CI =1.036-2.453, P=0.034); and 4) surgery plus adjuvant therapy subgroup (HR =1.976, 95% CI =1.226-3.185, P=0.005).However, the OS was not significantly correlated with OPRT or TP protein expression.

View Article: PubMed Central - PubMed

Affiliation: Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, People's Republic of China.

ABSTRACT
The aim of the present study was to investigate the clinicopathologic/prognostic significance of thymidylate synthase (TS), orotate phosphoribosyltransferase (OPRT), and thymidine phosphorylase (TP) proteins in postoperative non-small cell lung cancer (NSCLC) patients. Microarray slides from a set of 178 NSCLC patients were used for the detection of TS, OPRT, and TP expression by immunohistochemistry. The correlation between clinicopathologic factors and protein expression of three proteins was analyzed. Ninety seven carcinomas (57.4%) were TS-positive, 90 carcinomas (53.9%) were OPRT-positive, and 102 carcinomas (69.4%) were TP-positive. Compared with the TS-positive patients, the overall survival (OS) was significantly lower in the TS-negative patients (hazard ratio [HR] =1.766, 95% confidence interval [CI] =1.212-2.573, P=0.003). Significant differences between TS-positive and TS-negative patients was also observed in the following stratified analyses: 1) adenocarcinoma subgroup (HR =2.079, 95% CI =1.235-3.500, P=0.006); 2) less than 60-year-old subgroup (HR =1.890, 95% CI =1.061-3.366, P=0.031); 3) stage II/III subgroup (HR =1.594, 95% CI =1.036-2.453, P=0.034); and 4) surgery plus adjuvant therapy subgroup (HR =1.976, 95% CI =1.226-3.185, P=0.005). However, the OS was not significantly correlated with OPRT or TP protein expression. This study demonstrates that the TS level in tumor tissues may be a useful marker to predict the postoperative OS in NSCLC patients.

No MeSH data available.


Related in: MedlinePlus