World J Mens Health.  2013 Dec;31(3):247-253.

Expression of Heat Shock Protein 27 in Prostate Cancer Cell Lines According to the Extent of Malignancy and Doxazosin Treatment

• Lee SW
• Lee JW
• Chung JH
• Jo JK

Abstract

PURPOSE
Heat shock protein 27 (HSP27) is known as the material that plays a role in apoptosis control in tumor and cell protection including the immune response, drug tolerance, and so on. In this study, HSP27 expression according to the prostate cancer malignancy level was evaluated, and HSP27 expression was also analyzed after inducing apoptosis by doxazosin treatment of the prostate cancer cell lines.
MATERIALS AND METHODS
Reverse transcription polymerase chain reaction (RT-PCR) and immunofluorescence staining of the HSP27 was implemented by the culture of RWPE-1, LNCaP, androgen-independent human prostate cancer cells (PC-3), and TSU-Pr1 cell lines. Analysis was separately conducted in the control group, control vector group treated by dimethyl sulfoxide, and groups treated with 10 microM or 25 microM doxazosin. The expression of HSP27 in RT-PCR and immunofluorescence staining was observed and evaluated after conversion to numerical values.
RESULTS
In the RT-PCR results, depending on the cell type, LNCaP, TSU-Pr1 showed the highest HSP27 expression followed by PC-3, LNCaP and RWPE-1 in sequence. After doxazosin treatment, the expression detected by RT-PCR was stronger at a 25-microM doxazosin concentration compared to that at a 10-microM concentration, and the result was similar by immunofluorescence staining.
CONCLUSIONS
HSP27 expression increased depending on the prostate cancer cell line. This meant that HSP27 expression was related to the prostate cancer malignancy level. Additionally, the higher the treatment concentration in PC-3 was, the higher the HSP27 expression was. This result showed that doxazosin induced apoptosis of prostate cancer.

Keyword

Fluorescent antibody technique; Heat-shock proteins; Polymerase chain reaction; Reverse transcription

MeSH Terms

Apoptosis
Cell Line*
Control Groups
Cytoprotection
Dimethyl Sulfoxide
Doxazosin*
Drug Tolerance
Fluorescent Antibody Technique
Heat-Shock Proteins*
Hot Temperature*
HSP27 Heat-Shock Proteins*
Humans
Polymerase Chain Reaction
Prostatic Neoplasms
Reverse Transcription
Dimethyl Sulfoxide
Doxazosin
HSP27 Heat-Shock Proteins
Heat-Shock Proteins

Figure

• 

  Fig. 1 HSP27 reverse transcription polymerase chain reaction results according to the cell type (RWPE-1, LNCaP, PC-3, and TSU-Pr1). The density of HSP27 mRNA expression was shown to be higher than the low grade malignant cell line. Values are mean±standard error of mean. HSP27: heat shock protein 27, GAPDH: glyceraldehyde-3-phosphate dehydrogenase. ap<0.05 vs. control.

• 

  Fig. 2 Heat shock protein 27 immunofluorescence staining according to the cell type (RWPE-1, LNCaP, PC-3, and TSU-Pr1). The staining for TSU-Pr1 was more intense than that of the other cell lines. The intensity of staining was RWPE-1

• 

  Fig. 3 HSP27 gene expression patterns according to the doxazosin treatment concentration. The HSP27 gene expression with 25 µM of doxazosin treatment was higher than that with 10 µM (Control, CV, 10 µM, 25 µM). Values are mean±standard error of mean. HSP27: heat shock protein 27, GAPDH: glyceraldehyde-3-phosphate dehydrogenase. ap<0.05 vs. control.

• 

  Fig. 4 HSP27 immunofluorescence staining of PC-3 with doxazosin treatment (Control, CV, 10 µM, 25 µM). ×200; Fluoview confocal laser scanning microscope (Olympus Optical Co., Ltd., Tokyo, Japan). CV: control vehicle, HSP27: heat shock protein 27, TUNEL: terminal transferase-mediated biotinylated 16-desoxy-uridine triphosphate nick-end labeling.

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