Yonsei Med J.  2009 Jun;50(3):399-406. 10.3349/ymj.2009.50.3.399.

Expression of Heat Shock Protein 60 kDa Is Upregulated in Cervical Cancer

• Hwang YJ
• Lee SP
• Kim SY
• Choi YH
• Kim MJ
• Lee CH
• Lee JY
• Kim DY

• KMID: 1758569
• DOI: http://doi.org/10.3349/ymj.2009.50.3.399

Abstract

PURPOSE: Cervical cancer caused by the human papilloma virus (HPV) continues to be the cause of yearly death among women. However, it is a curable disease when diagnosed at an early stage. Recently, several researches have reported that heat shock protein (HSP) 60, a chaperone protein of molecular weight of 60 kDa, is involved in carcinogenesis and apoptosis. In order to evaluate the prognostic significance of HSP60 in cervical cancer, we examined differences in the HSP60 expression between cervical cancer and normal tissues in women.
MATERIALS AND METHODS
Tissue samples were collected from 20 cervical cancer patients and 20 normal controls. HSP60 expression of cervical cancer and normal tissues were verified by the 2D gel proteomics, semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot analyses.
RESULTS
In 2D proteomic analysis, an increase of HSP60 expression was detected in cervical cancer tissues and confirmed by Western blot analysis (p < 0.05). However, messenger RNA (mRNA) levels of HSP60 did not display any significant differences between cervical cancer and normal tissues.
CONCLUSION
These results suggest that HSP60 may be involved in the development of cervical cancer and have profound biological and prognostic significance.

Keyword

Heak shock protein; cervical cancer; prognostic marker

MeSH Terms

Adult
Aged
Blotting, Western
Chaperonin 60/*metabolism
Electrophoresis, Gel, Two-Dimensional
Female
Humans
Male
Middle Aged
Reverse Transcriptase Polymerase Chain Reaction
Uterine Cervical Neoplasms/*metabolism

Figure

• 

  Fig. 1 Proteome pattern of normal cervical tissue. Eleven protein spots on the gel were marked with arrows. Numbered spots were excised from the normal tissue gel, in-gel digested with trypsin, and identified by MALDI-TOF assay. The results are listed in Table 1. MALDI-TOF, matrix assisted laser desorption ionization-Time of flight mass spectrometry.

• 

  Fig. 2 Proteome pattern of cervical cancer tissue. Nineteen protein spots on the gel were marked with arrows. Numbered spots were excised from the cancer tissue gel, in-gel digested with trypsin, and identified by MALDI-TOF assay. The results are listed in Table 2. MALDI-TOF, matrix assisted laser desorption ionization-Time of flight mass spectrometry.

• 

  Fig. 3 (A) RT-PCR analysis of HSP60 mRNA in normal (lane 1-8) and cervical cancer (lane 9-16) tissues. (B) RT-PCR was performed using 1 µg of total RNA and separated on 2.5% agarose gel. The size of PCR products was 320 base pairs. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal control to confirm equal loading of the samples. HSP60 mRNA levels were quantified as a percentage of relative optical density. Results are mean ± S.E.M. of 20 samples per group. RT-PCR, reverse transcriptase polymerase chain reaction; mRNA, messenger RNA; HSP60, heat shock protein.

• 

  Fig. 4 (A) Western blot analysis of HSP60 protein in normal controls (lane N1-N20) and cervical cancers (lane C1-C20) tissues. (B) Anti-actin protein was used as a control. HSP60 protein levels were quantified as a percentage of relative optical density. Results are mean ± S.E.M. of 20 samples per group. Expression level of HSP60 protein in cervical cancer tissues was significantly higher than in normal tissues (p < 0.05). HSP60, heat shock protein.

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