Tuberc Respir Dis.  2013 Aug;75(2):59-66.

Elevated Prx1 Provides Resistance to Docetaxel, But Is Not Associated with Predictive Significance in Lung Cancer

• Hwang KE
• Park C
• Seol CH
• Hwang YR
• Hwang JS
• Jung JW
• Choi KH
• Jeong ET
• Kim HR

Abstract

BACKGROUND
This study was conducted in order to elucidate the effects of docetaxel on the growth of peroxiredoxin 1 (Prx1) knockdown A549 xenograft tumors and further tested the role of Prx1 as a predictor for how a patient would respond to docetaxel treatment.
METHODS
Effects of docetaxel on the growth of scrambled- and shPrx1-infected A549 xenograft tumors in nude mice were measured. Moreover, immunohistochemical expression of Prx1 was evaluated in paraffin-embedded tissues from 24 non-small cell lung cancer patients who had received docetaxel-cisplatin regimens as a first-line treatment.
RESULTS
Docetaxel treatment in Prx1 knockdown xenograft tumor resulted in reduced tumors growth compared with other groups. Prx1 knockdown increased the production of cleaved caspases-8 and -9 in the control itself compared to scramble tumors. Moreover, docetaxel treatment in Prx1 knockdown tissue led to an increased protein band. Phosphorylated Akt was found in Prx1 scramble tissues. Phosphorylated FOXO1 was detected in the docetaxel treatment group. On the other hand, Prx1 knockdown completely suppressed the Akt-FOXO1 axis. The median progression-free survival (PFS) of patients with low Prx1 expression was 7 months (95% confidence interval [CI], 6.0-7.7), whereas the median progression-free survival of patients with high Prx1 expression was 4 months (95% CI, 4.0-5.0). However, high Prx1 expression was not associated with decreased PFS (p=0.114).
CONCLUSION
Our findings suggest that elevated Prx1 provides resistance to docetaxel treatment through suppression of FOXO1-induced apoptosis in A549 xenograft tumors, but may not be related with the predictive significance for response to docetaxel treatment.

Keyword

PRDX1 Protein, Human; Docetaxel; Lung Neoplasms

MeSH Terms

Animals
Apoptosis
Axis, Cervical Vertebra
Carcinoma, Non-Small-Cell Lung
Disease-Free Survival
Hand
Humans
Lung
Lung Neoplasms
Mice
Mice, Nude
Peroxiredoxins
Taxoids
Transplantation, Heterologous
Peroxiredoxins
Taxoids

Figure

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  Figure 1 Peroxiredoxin 1 (Prx1) knockdown inhibited the growth of A549 tumors in docetaxel-treated mice. Athymic nude mice were injected subcutaneously with 2×106 scrambled- or shPrx1-infected A549 cells (0.2 mL cell suspension) in both hind legs. The mice were randomly divided into 4 groups of 7 animals each: group 1, mice bearing scrambled-infected tumors and receiving human IgG treatment, serving as control for group 2; group 2, mice bearing scrambled-infected tumors and receiving docetaxel treatment at 20 mg/kg, intraperitoneally, once a week; group 3, mice bearing shPrx1-infected tumors and receiving human IgG treatment, serving as control for group 4; and group 4, mice bearing shPrx1-infected tumors and receiving docetaxel treatment at 20 mg/kg, i.p., once a week. Tumors were measured 3 times per week. Tumor volume was estimated using the formula: volume=L×W2/2. Points, mean of 7 animals; bars, standard deviation.

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  Figure 2 Peroxiredoxin 1 (Prx1) knockdown suppressed docetaxel-induced apoptosis in A549 xenograft tumors. Tumors were analyzed for apoptotic proteins by using immunoblot assay. Cell lysates were subjected to 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis to measure the expression of caspase-8, -9, and Prx1. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the protein loading control.

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  Figure 3 Peroxiredoxin 1 (Prx1) knockdown suppressed docetaxel-induced Akt-FOXO1 pathways in A549 xenograft tumors. Tumors were analyzed for Akt-FOXO1 proteins by using the immunoblot assay. Cell lysates were subjected to 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis to measure the expression of phosphorylated Akt and FOXO1. The same membrane used for anti-phospho antibody staining was stripped and used again with antibody for total Akt. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the protein loading control.

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  Figure 4 Peroxiredoxin 1 (Prx1) knockdown activated FOXO1 induced TRAIL, but not FasL. Tumors were analyzed for death receptors by using reverse transcription polymerase chain reaction (RT-PCR), which was performed to amplify the TRAIL and FasL genes as described in Materials and Methods. Equal mRNA and protein loading were confirmed using β-actin. RT-PCR data are representative of at least 2 independent experiments.

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  Figure 5 High peroxiredoxin 1 (Prx1) expression did not correlate with progression-free survival (PFS). (A) Immunohistochemical expression of Prx1 in non-small cell lung carcinoma. Typical immunohistological features with high levels of Prx1 expression in squamous cell carcinoma and adenocarcinoma (×200). The Prx1 staining was primarily present in the cytoplasm of tumor cells. Focal nuclear staining was observed in some malignant cells. (B) PFS determined using Kaplan-Meier survival analysis. The p-value was determined from a log-rank test of the difference.

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