Ubiquitous mitochondrial creatine kinase downregulated in oral squamous cell carcinoma

Onda, Takeshi; Uzawa, Katsuhiro; Endo, Yutaka; Bukawa, Hiroki; Yokoe, Hidetaka; Shibahara, Takahiko; Tanzawa, Hideki

doi:10.1038/sj.bjc.6602986

Cited by 61 publications

(56 citation statements)

References 49 publications

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“…The slides were then lightly counterstained with hematoxylin, dehydrated with ethanol, cleaned with xylene and mounted. To quantify the state of FHL1 protein expression in those components, we used IHC score systems described previously (18,(22)(23)(24)(25)(26)(27)(28)(29)(30). Briefly, the stained cells were determined in at least five random fields at magnification x400 in each section.…”

Section: Methodsmentioning

confidence: 99%

High prevalence of epigenetic inactivation of the human four and a half LIM domains 1 gene in human oral cancer

Koike

Kasamatsu

Iyoda

et al. 2012

International Journal of Oncology

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Abstract.The four and a half LIM domains 1 (FHL1) gene has been related to carcinogenesis. However, the expression status of FHL1 in human oral squamous cell carcinoma (OSCC) remains unclear and the detailed mechanism of gene silencing is poorly understood. The aim of this study was to examine the FHL1 expression level and its regulatory mechanism in OSCCs. Quantitative reverse-transcriptase-polymerase chain reaction (PCR) and western blotting showed significant downregulation of FHL1 in all OSCC-derived cell lines (Sa3, HSC-2, HSC-3, HSC-4, HO-1-u-1, HO-1-N-1, KON and Ca9-22) compared to human normal oral keratinocytes. We also found that FHL1 mRNA expression was frequently downregulated (P<0.01) in 51 (86.4%) of 59 primary OSCCs compared with the corresponding normal oral tissues, while there was no significant difference between the status of the FHL1 protein expression in OSCCs and the clinicopathological features. Using methylationspecific PCR, we detected methylated FHL1 in all cell lines and treatment with the DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine restored the FHL1 expression. However, no significant restoration of FHL1 expression was observed using sodium butyrate, an inhibitor of histone deacetylase and chromatin immunoprecipitation showed that histone H3 lysine 9 in the FHL1 promoter region was significantly acetylated. In addition, no mutation in the entire coding region of the FHL1 gene was found. Therefore, our data suggested that inactivation of the FHL1 gene is a frequent event during oral carcinogenesis and that the mechanism of FHL1 downregulation in OSCCs is through DNA methylation of the promoter region rather than histone deacetylation or mutation. IntroductionHead and neck squamous cell carcinoma (HNSCC) is one of the most frequently occurring malignancies and a major cause of morbidity and mortality (1,2). Oral cancer is the most common among the HNSCCs and the most frequently occurring oral cancer is squamous cell carcinoma (OSCC), which accounts for more than 90% of all oral malignancies (3). The number of OSCC cases that occur worldwide annually exceeds 300,000 (4). The mechanisms behind tumor progression of OSCC are known to a limited extent, indicating a clear need for comprehensive knowledge that can lead to more specific and effective molecular target.Microarray technology facilitates simultaneous evaluation of thousands of genes in a specimen. The results of microarray analysis provide researchers with high throughput screening to study the roles played by specific genes in cancer development and progression. We previously reported gene expression profiling of OSCC to identify cancer-related genes (5,6).The four and a half LIM domains (FHL) family of genes is characterized by LIM domains, a term derived from the first letters of three transcription factors: Lin-11, Isl-1 and Mec-3. Because the LIM domains provide protein-protein binding interfaces, the FHL genes play an important role in cellular events such as focal adhesion and differentiation by repressing or ac...

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Section: Methodsmentioning

confidence: 99%

High prevalence of epigenetic inactivation of the human four and a half LIM domains 1 gene in human oral cancer

Koike

Kasamatsu

Iyoda

et al. 2012

International Journal of Oncology

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“…As a negative control, triplicate sections were immunostained without exposure to primary antibodies, which confirmed the staining specificity. To quantify the state of HSF1 protein expression in subcellular fractionations (nucleus, nucleus plus cytoplasm and cytoplasm), we used IHC score systems described previously (22,(24)(25)(26)(27)(28)(29)(30)(31)(32)(33). Briefly, the stained cells were determined in at least five random fields at x400 magnification in each section.…”

Section: Introductionmentioning

confidence: 99%

State of heat shock factor 1 expression as a putative diagnostic marker for oral squamous cell carcinoma

et al. 2011

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Abstract. Heat shock factor 1 (HSF1) is responsible for expression of a large class of heat shock proteins that have been implicated in the malignant phenotype of human cancers. Little is known about the effect of a high level of HSF1 on the behavior of oral squamous cell carcinoma (OSCC). In this study, we assessed the value of HSF1 for predicting clinical outcomes in OSCC. Quantitative reverse transcriptase-polymerase chain reaction and Western blotting showed that the expressions of HSF1 mRNA and protein in OSCC-derived cell lines (HSC-2, HSC-3, HSC-4, Sa3, Ca9-22, KON and Ho-1-u-1) were elevated compared with those in human normal oral keratinocytes (P<0.05). Similar to in vitro data, HSF1 mRNA expression in primary OSCCs (n=50) was significantly greater than in normal counterparts (P<0.05). Since HSF1 was observed in the nucleus and cytoplasm by immunohistochemistry, we investigated the correlation between the HSF1 expression status at each subcellular location and the clinical behavior of OSCCs. Among the clinical classifications, higher nuclear HSF1 expression was closely related to tumor size and histopathologic types (P<0.05). These results showed for the first time that nuclear HSF1 expression may contribute to cancer progression and that HSF1 might be a potential diagnostic biomarker and a therapeutic target for

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“…All cell lines were maintained at 37˚C (humidified atmosphere 5% CO 2 /95% air) on 150x20 mm tissue culture dishes (Nunc, Roskilde, Denmark) and cultured in Dulbecco's modified Eagle's medium F-12 HAM (Sigma, St. Louis, MO, USA) with 10% fetal bovine serum (Sigma) plus 50 U/ml penicillin and streptomycin (16).…”

Section: Cellsmentioning

confidence: 99%

“…The reaction mixture was loaded into glass capillary tubes and submitted to an initial denaturation at 95˚C for 10 min, followed by 45 rounds of amplification at 95˚C (10 sec) for denaturation, 56˚C (10 sec) for annealing, and 72˚C for extension, with a temperature slope of 20˚C, performed in the LightCycler. The amount of transcript from the FAM5C gene was estimated from the respective standard curves and normalized to the amount of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) transcript determined in corresponding samples (16). …”

Section: '-Aggacaccacaagttccagg-3' ----------------------------------mentioning

confidence: 99%

Expression of the FAM5C in tongue squamous cell carcinoma

Kuroiwa

Yamamoto²,

Onda³

et al. 2009

Oncol Rep

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Abstract. The purpose of this study was to perform a wholegenome 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 wholegenome 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.

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Ubiquitous mitochondrial creatine kinase downregulated in oral squamous cell carcinoma

Cited by 61 publications

References 49 publications

High prevalence of epigenetic inactivation of the human four and a half LIM domains 1 gene in human oral cancer

High prevalence of epigenetic inactivation of the human four and a half LIM domains 1 gene in human oral cancer

State of heat shock factor 1 expression as a putative diagnostic marker for oral squamous cell carcinoma

Expression of the FAM5C in tongue squamous cell carcinoma

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