Yonsei Med J.  2011 May;52(3):463-468. 10.3349/ymj.2011.52.3.463.

Gene Therapy for Head and Neck Squamous Cell Carcinoma Using KITENIN (KAI1 COOH-Terminal Interacting Tetraspanin)-Antisense Therapy

• Lee JK
• Lee DH
• Sun EG
• Bae JA
• Lim SC
• Min JJ
• Sung MW
• Kim KK

• KMID: 1777018
• DOI: http://doi.org/10.3349/ymj.2011.52.3.463

Abstract

PURPOSE
KAI1 COOH-terminal interacting tetraspanin (KITENIN) has been found to act as a promoter of metastasis in murine models of colon cancer and squamous cell carcinoma (SCC). The suppression of tumor progression and metastasis of established colon cancer in mice was observed after intravenous delivery of small interfering RNA (siRNA) targeting KITENIN. The purpose of this study was to investigate the efficacy of gene therapy targeting KITENIN in human head and neck SCC.
MATERIALS AND METHODS
SNU-1041, a well-established human hypopharyngeal SCC cell line, was used. KITENIN expression in SNU-1041 was measured by Western blot analysis. The cells were prepared, maintained in culture dishes with media, and divided into two groups: the si-KITENIN group and the scrambled group (control). The siRNA targeting KITENIN (si-KITENIN) and scrambled DNA were transfected into the SNU-1041 cells in each group. The effect of gene therapy was compared by in vitro experiments to evaluate invasion, migration, and proliferation.
RESULTS
KITENIN was strongly expressed in the SNU-1041 cells, and the number of invaded cells was reduced more in the si-KITENIN group than in the scrambled group (p<0.001). The speed for the narrowing gap, made through adherent cells, was lower in the si-KITENIN group (p<0.001), and the number of viable proliferating cells was reduced in the si-KITENIN group compared to the scrambled group (p<0.001, the third day). KITENIN protein expression was no longer identified in the si-KITENIN group.
CONCLUSION
Gene therapy using an anti-KITENIN strategy might be effective for head and neck squamous carcinoma.

Keyword

Gene therapy; head and neck cancer; squamous cell carcinoma; KITENIN protein; small interfering RNA (siRNA)

MeSH Terms

Carcinoma, Squamous Cell/genetics/pathology/*therapy
Carrier Proteins/*antagonists & inhibitors/genetics
Cell Line, Tumor
Cell Movement
Cell Proliferation
*Gene Therapy
Head and Neck Neoplasms/genetics/pathology/*therapy
Humans
Membrane Proteins/*antagonists & inhibitors/genetics
*RNA, Small Interfering

Figure

• 

  Fig. 1 KITENIN expression in SNU-1041 cells: KITENIN was strongly expressed in SNU-1041 cells and scrambled DNA-transfected SNU-1041 cells (SNU-1041/Scrambled). The expression of KITENIN protein was not observed in si-KITENIN-transfected SNU-1041 cells (SNU-1041/si-KITENIN).

• 

  Fig. 2 Cell invasion assay: the number of invading si-KITENIN-transfected SNU-1041 cells (A) was 64.2±19.2, whereas the number was 98.6±10.2 for the scrambled DNA-transfected SNU-1041 cells (B), as measured by 10 random squares of 0.5×0.5 mm2 on the microscopic field of view under conditions with 20 µg/mL of fibronectin; the difference between the two groups was statistically significant (p<0.001)(C).

• 

  Fig. 3 Cell migration assay: the artificial wound gap became significantly narrower in the control group as time passed by 20, 28, 44 hours, compared to the si-KITENIN group (p<0.001)(B). In the scrambled group, the gap was nearly filled within 44 hours. However, it remained wide open in the si-KITENIN group (A).

• 

  Fig. 4 Cell proliferation assay: the proliferating cells, as determined by absorbance, were significantly decreased in the si-KITENIN group compared to the scrambled group. On the third day, absorbance was 0.4933±0.0484 in the si-KITENIN group and 0.6023±0.0319 in the scrambled group (p<0.001).

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