Tetraspanin gene expression levels as potential biomarkers for malignancy of gingival squamous cell carcinoma

Abstract

Accurate assessment of malignancy in oral squamous cell carcinoma is essential to optimize treatment planning. To detect a biomarker related to malignant propensity in gingival squamous cell carcinoma (GSCC), quantitative gene expression analysis of tetraspanin family genes was conducted. In 73 cases of GSCC, total RNA was extracted from carcinoma tissues, and gene expression was analyzed by quantitative real time-PCR. Six tetraspanin family genes (CD9, CD63, CD81, CD82, CD151, NAG-2) were investigated. Housekeeping genes (ACTB and GAPDH), anchor protein genes (JUP and PXN) and an integrin gene (ITGA3) were used as reference genes. Forty-five gene expression ratios were calculated from these 11 gene expression levels and were analyzed with clinical parameters using multivariate statistical methods. According to the results of the logistic regression analysis subjecting cervical lymph node metastasis as a target variable, CD9/ACTB (p 5 0.013) or CD9/CD82 (p 5 0.013) in addition to tumor size (p 5 0.028) were detected as significant factors. In Cox proportional hazards regression analysis, delayed cervical lymph node metastasis (p 5 0.039) and tumor cell positive surgical margin (p 5 0.032) in addition to CD151/GAPDH (p 5 0.024) were detected as significant factors for death outcome. A Kaplan-Meier survival curve presented a significantly lower survival rate of the group with a CD151/GAPDH value of 10 or more (log rank and generalized Wilcoxon tests: p 5 0.0003). Results of this study present the usefulness of CD9 and CD151 expression levels as biomarkers for assessment of malignancy in GSCC. They also indicate that detection of residual tumor cells at the surgical margin and the biological malignancy of a tumor interdependently affects prognosis. ' 2009 UICC Key words: biomarker; tetraspanin; CD9; CD82; CD151; gingival squamous cell carcinoma; lymph node metastasis; death outcomeThe incidence of squamous cell carcinoma (SCC) arising from gingiva is relatively high among oral malignant neoplasms, and approximately half of the cases also presented with cervical lymph node (LN) metastasis. Because gingival SCC (GSCC) easily invades the jawbone and its surrounding tissues such as nerve, muscles, the nasal cavity and skin, determination of the extent of resection and reconstruction are significant issues in treatment of advanced cases.1-3 Therefore, determination of optimal treatment plans based on accurate assessment of biological malignancy is extremely important not only in terms of controlling cancers but also for minimizing dysfunction after treatment. Candidates for biomarkers of head and neck cancers including oral SCC (OSCC) were found in some studies; however, they have not been of practical use. 4,5 Although diagnostic imaging, status of lymph node metastasis and histology still will be clinically essential criteria, they are not able to provide sufficient information to realize the individualized treatment of OSCC in the future. A precise malignancy diagnosis of OSCC, such as prediction of potential metast...

ITGA3 and ITGB4 expression biomarkers estimate the risks of locoregional and hematogenous dissemination of oral squamous cell carcinoma

“…In our previous study, we analyzed the ratio of expression of the 11 ITG family genes to that of the 14 functionally related genes; in total, 154 gene expression ratios for 66 tongue SCC cases [15]. We also investigated the potential of 45 tetraspanin family gene expression ratios that were calculated based on 6 tetraspanin family genes with housekeeping functionality or functionally related genes for 73 gingival SCC cases [16]. The results of these prior studies revealed two ITG gene expression ratios–those of ITGA3/CD9 and ITGB4/JUP–as candidate biomarkers for OSCC.…”

“…The α3β1 integrin complex is a major receptor for laminin 5 [17] and is involved in the maintenance of epithelial integrity, cell proliferation and motility, and survival of migrating keratinocytes through adhesion to extracellular matrix components [21-23]. CD9 is a tetraspanin family molecule, which forms tetraspanin webs (tetraspanin-enriched microdomains) by associating with various partner molecules such as integrins, growth factor receptors, and other tetraspanin molecules, to affect cell adhesion, signal transduction, proliferation, motility, differentiation, and cancer metastasis [16,24-28]. It has been reported that CD9 negatively influences cancer cell motility by regulating the re-organization of the actin cytoskeleton [29].…”

“…In our previous studies, we used microarray analysis and reverse transcription quantitative real time polymerase chain reaction (RT-QPCR) to report the potential use of integrin and tetraspanin family molecules as biomarkers for OSCC malignancy [14-16]. The integrin (ITG) molecule functions as a cell surface receptor that mediates extracellular mechanical and chemical signals into the cell interior, which modulates different signal transduction cascades.…”

Tetraspanins CD9 and CD151, epidermal growth factor receptor and cyclooxygenase-2 expression predict malignant progression in oral epithelial dysplasia

“…Three of the tetraspanins (CD9, CD151, and CD82) have been shown to interact with EGFR, either directly or in conjunction with integrin adhesion receptors (Wang et al , 2007; Park et al , 2009; Deng et al , 2012; Romanska et al , 2012; Odintsova et al , 2013). Furthermore, there is evidence that reduced expression of CD9 (Mhawech et al , 2004; Buim et al , 2010) and CD82 (Imai et al , 2002; Farhadieh et al , 2004), and increased expression of CD151 (Hirano et al , 2009) confers a poor prognosis in head and neck cancer. It is possible, therefore, that these tetraspanins may also have a role earlier in the carcinogenic process.…”

CD151 (CD151 molecule (Raph blood group))

“…Note Real-time PCR analysis of CD151 gene expression compared to GAPDH was analysed (Hirano et al, 2009). Assessment of protein expression by immunohistochemistry correlated with gene expression however no statistical analyses were performed on protein expression.…”