J Gynecol Oncol.  2010 Mar;21(1):29-37. 10.3802/jgo.2010.21.1.29.

Expression pattern of the class I homeobox genes in ovarian carcinoma

• Hong JH
• Lee JK
• Park JJ
• Lee NW
• Lee KW
• Na JY

• KMID: 2173508
• DOI: http://doi.org/10.3802/jgo.2010.21.1.29

Abstract

OBJECTIVE
Although some sporadic reports reveal the link between the homeobox (HOX) genes and ovarian carcinoma, there is no comprehensive analysis of the expression pattern of the class I homeobox genes in ovarian carcinoma that determines the candidate genes involved in ovarian carcinogenesis. METHODS: The different patterns of expression of 36 HOX genes were analyzed, including 4 ovarian cancer cell lines and 4 normal ovarian tissues. Using a reverse transcription-polymerase chain reaction (RT-PCR) and quantification analysis, the specific gene that showed a significantly higher expression in ovarian cancer cell lines than in normal ovaries was selected, and western blot analysis was performed adding 7 ovarian cancer tissue specimens. Finally, immunohistochemical and immunocytochemical analyses were performed to compare the pattern of expression of the specific HOX gene between ovarian cancer tissue and normal ovaries. RESULTS: Among 36 genes, 11 genes had a different level of mRNA expression between the cancer cell lines and the normal ovarian tissues. Of the 11 genes, only HOXB4 had a significantly higher level of expression in ovarian cancer cell lines than in normal ovaries (p=0.029). Based on western blot, immunohistochemical, and immunocytochemical analyses, HOXB4 was expressed exclusively in the ovarian cancer cell lines or cancer tissue specimens, but not in the normal ovaries. CONCLUSION: We suggest HOXB4 may be a novel candidate gene involved in ovarian carcinogenesis.

Keyword

Homeobox gene; Ovarian neoplasms; Carcinogenesis

MeSH Terms

Aluminum Hydroxide
Blotting, Western
Carbonates
Cell Line
Female
Genes, Homeobox
Ovarian Neoplasms
Ovary
RNA, Messenger
Aluminum Hydroxide
Carbonates
RNA, Messenger

Figure

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  Fig. 1 RT-PCR results of 36 homeobox (HOX) genes in 4 ovarian cancer cell lines and 4 normal ovarian tissues. (A) Reverse transcription-polymerase chain reaction (RT-PCR) results of 10 HOXA and 8 HOXB genes. (B) RT-PCR results of 9 HOXC and 9 HOXD genes. C: ovarian cancer cell line, C1: SK-OV3, C2: TOV-21G, C3: SW626, C4: OV-90, N 1-4: normal ovarian tissues.

• 

  Fig. 2 A Expression pattern of homeobox B4 (HOXB4) by reverse transcription-polymerase chain reaction (RT-PCR) in normal ovaries and ovarian cancer cell lines. (A) HOX B4. (B) GAPDH B quantification analysis of HOX B4. HOX B4 expression was significantly higher in ovarian cancer cell lines than in normal ovaries (*p=0.029). N 1-4: normal ovarian tissues, C: ovarian cancer cell line, C1: OV-90, C2: SW-626, C3: TOV-21G, C4: SK-OV3.

• 

  Fig. 3 Detection of homeobox B4 (HOXB4) by western blot in the 4 cancer cell lines, 7 cancer tissue specimens and 4 normal ovaries. 1: SK-OV3, 2: TOV-21G, 3: SW 626, 4: OV-90, 5-11: ovarian cancer tissues, 12-15: normal ovarian tissues.

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  Fig. 4 Immunohistochemical staining of homeobox B4 (HOXB4) in ovarian cancer tissue (A) and normal ovarian tissue (B). (A) Mainly cytoplasmic with some perinuclear staining was seen exclusively in the epithelial cell, not in the stroma. All of the ovarian cancer tissue was serous in histology (×400). (B) No anti-HOXB4 antibody activity in the normal ovarian epithelial cell (×100).

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  Fig. 5 Immunocytochemical results of homeobox B4 (HOXB4) in ovarian cancer cell lines and normal ovarian epithelium. Strong cytoplasmic staining was seen in all 4 ovarian cancer cell lines. In contrast, no staining was observed in the normal ovarian epithelial cell. (A) SK-OV3, (B) TOV-21G, (C) SW 626, (D) OV-90, (E) Normal ovarian epithelium. (A~D) ×400, (E) ×100.

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