Kuroiwa Tsukasa, Yamamoto Nobuharu, Onda Takeshi, Shibahara Takahiko
Department of Oral and Maxillo-Facial Surgery, Tokyo Dental College, Chiba 261-8502, Japan.
Oncol Rep. 2009 Nov;22(5):1005-11. doi: 10.3892/or_00000528.
The purpose of this study was to perform a whole-genome analysis of loss of heterozygosity (LOH) in tongue squamous cell carcinoma (SCC) using the Affymetrix 10K SNP Mapping Array. In the gene which had been identified by whole-genome analysis of LOH, we analyzed allelic imbalance to identify the role of the gene. We applied whole-genome analysis of LOH in the specimens from the 5 cases of tongue SCC using this array. In the chromosomal region which had been identified by whole-genome analysis of LOH, we reconfirmed the existence of LOH in 30 cases using microsatellite markers. The expression levels of the mRNA in the region were examined in 15 cases and in 5 tongue SCC-derived cell lines by real-time quantitative RT-PCR analysis. LOH was observed in all of the 5 cases in the 1q31.1 region. Only 3 microsatellite markers (D1S1189, D1S2151, and D1S2595) existed in the 1q31.1 region. A high frequency of LOH was found at the D1S1189 locus in 18/30 (60%), D1S2151 locus in 16/30 (53%) and D1S2595 locus in 21/30 (70%). Only the Family with sequence similarity 5, member C (FAM5C) gene was located in the 1q31.1 region. There was statistically significant difference in the FAM5C mRNA expression levels between tongue SCC and normal tissues. All tongue SCC-derived cell lines decreased FAM5C mRNA expression compared with normal oral keratinocytes (NOKs). We conclude that FAM5C may be a novel tumor suppressor gene (TSG) in tongue SCC.
本研究的目的是使用Affymetrix 10K SNP映射阵列对舌鳞状细胞癌(SCC)中的杂合性缺失(LOH)进行全基因组分析。在通过LOH全基因组分析鉴定出的基因中,我们分析了等位基因失衡以确定该基因的作用。我们使用该阵列对5例舌SCC标本进行了LOH全基因组分析。在通过LOH全基因组分析鉴定出的染色体区域中,我们使用微卫星标记在30例中重新确认了LOH的存在。通过实时定量RT-PCR分析检测了15例和5种舌SCC衍生细胞系中该区域mRNA的表达水平。在1q31.1区域的所有5例中均观察到LOH。1q31.1区域仅存在3个微卫星标记(D1S1189、D1S2151和D1S2595)。在D1S1189位点,18/30(60%)出现高频率的LOH;在D1S2151位点,16/30(53%)出现高频率的LOH;在D1S2595位点,21/30(70%)出现高频率的LOH。只有序列相似性家族5成员C(FAM5C)基因位于1q31.1区域。舌SCC与正常组织之间FAM5C mRNA表达水平存在统计学显著差异。与正常口腔角质形成细胞(NOKs)相比,所有舌SCC衍生细胞系的FAM5C mRNA表达均降低。我们得出结论,FAM5C可能是舌SCC中的一种新型肿瘤抑制基因(TSG)。