Global Expression-Based Classification of Lymph Node Metastasis and Extracapsular Spread of Oral Tongue Squamous Cell Carcinoma

Abstract

Regional lymph node metastasis is a critical event in oral tongue squamous cell carcinoma (OTSCC) progression. The identification of biomarkers associated with the metastatic process would provide critical prognostic information to facilitate clinical decision making for improved management of OTSCC patients. Global expressional profiles were obtained for 25 primary OTSCCs, where 11 cases showed lymph node metastasis (pN+) histologically and 14 cases were nonmetastatic (pN-). Seven of pN+ cases also exhibited extracapsular spread (ECS) of metastatic nodes. Multiple expression indices were used to generate signature gene sets for pN+/- and ECS+/- cases. Selected genes from signature gene sets were validated using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The classification powers of these genes were then evaluated using a logistic model, receiver operating characteristic curve analysis, and leave-one-out cross-validation. qRT-PCR validation data showed that differences at RNA levels are either statistically significant (P < .05) or suggestive (P < .1) for six of eight genes tested (BMP2, CTTN, EEF1A1, GTSE1, MMP9, and EGFR) for pN+/- cases, and for five of eight genes tested (BMP2, CTTN, EEF1A1, MMP9, and EGFR) for ECS+/- cases. Logistic models with specific combinations of genes (CTTN+MMP9+EGFR for pN and CTTN+EEF1A1+MMP9 for ECS) achieved perfect specificity and sensitivity. Leave-one-out cross-validation showed overall accuracy rates of 85% for both pN and ECS prediction models. Our results demonstrated that the pN and the ECS of OTSCCs can be predicted by gene expression analyses of primary tumors.

SERPINE1 and SMA expression at the invasive front predict extracapsular spread and survival in oral squamous cell carcinoma

“…This supports our hypothesis that molecular determinants of ECS are present in the primary tumour. In a recent meta-analysis of over 60 studies of differential gene-expression profiling in HNSCC, one study of a limited number of cases predicted ECS by geneexpression analysis from primary site tissue (Zhou et al, 2006;Yu et al, 2008). It is also evident from many studies that there is a high risk for false discovery, and careful validation is mandated before establishing clinical biomarkers (Mroz and Rocco, 2012;van Hooff et al, 2012a).…”

Genomic assessments of the frequent loss of heterozygosity region on 8p21.3∼p22 in head and neck squamous cell carcinoma

“…To evaluate the expressional changes of MTUS1 genes at mRNA level, the quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed as described previously [15] on 10 HNSCC cell lines and normal oral keratinocyte (NHOK), as well as 10 paired normal and oral tongue cancer tissues samples (see Supplement Table 2 for the descriptions of the oral tongue cancer tissues). The RNA from cell lines was isolated using Qiagen RNeasy kit.…”

“…Mitochondrial tumor suppressor gene 1 (MTUS1) is one of the candidate genes that reside in this genomic region, which was initially identified as a potential tumor suppressor gene in pancreatic cancer [14]. Our recent study suggested that the reduction of MTUS1 expression may be associated with advanced oral tongue SCC [15]. The reduced expression of MTUS1 has also been observed in colon cancer, ovarian cancer and pancreatic cancer [14,16,17].…”

Downregulation of HOPX Controls Metastatic Behavior in Sarcoma Cells and Identifies Genes Associated with Metastasis

“…We subsequently validated the microarray results by qRT-PCR of several genes, ADCY7, ARGHDIB, BAMBI, FOXG1, FSCN1, FUCA1, GFPT2, ITPR3, ITGA4, LSP1, NCAM1, NUDT14, RGS20, RHOF, and RIAM, that have already been implicated in cell migration, adhesion, or angiogenesis (28,(33)(34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44)(45)(46). The results from qRT-PCR analysis (Fig.…”