Yuk Kwan Chen

Primary oral squamous cell carcinoma: an analysis of 703 cases in southern Taiwan.

We retrospectively analyzed the records of 703 cases of oral squamous cell carcinoma (SCC) collected from 1 January 1985 to 31 December 1996 at a teaching hospital in southern Taiwan, to identify the characteristics of patients and factors associated with survival. There was an overwhelming male predominance (male:female = 15:1). The mean age of the patients was 52. The peak age of oral SCC patients declined from 50 to 59 years in the first six years (1985-1990) and 40-49 years in the last six years (1991-1996). The most common site of oral SCC was the buccal mucosa with 263 patients (37.4%). Most patients (346/703 patients; 49.2%) had stage III cancer. The most common site of occurrence of SCC was the buccal mucosa (263/703 patients; 37.4%), both overall and in patients who chewed betel quid alone or in combination with cigarette smoking and/or alcohol consumption; the tongue was the most common site among patients without any oral habits (18/48 patients; 37.5%). Furthermore, the age of occurrence was on average 6-12 years younger among patients who chewed betel quid than in those who did not. Of the 703 patients, 496 received treatment with surgery, chemotherapy, and/or radiation therapy. Of these, 209 (42.1%) died. The cancer stage significantly influenced mortality: the 5-year survival rate in patients treated from 1985 to 1991 was 72% in those with stage I, 38.9% in those with stage II, 26.7% in those with stage III, and 11.8% in those with stage IV cancer. Six variables were found to significantly affect survival: tumor size, lymph node involvement, surgery, betel quid chewing, staging, and histological differentiation (all p < 0.05, Kaplan-Meier analysis with log rank test). Of these, surgery and cancer stage independently affected survival in a proportional hazards model (both p < 0.0001). Therefore, the early surgical intervention, and the withdrawal from oral habits, especially betel quid chewing, will be advantageous to patients survival.

Malignant transformation in 5071 southern Taiwanese patients with potentially malignant oral mucosal disorders.

BackgroundOral cancers can be preceded by clinically evident oral potentially malignant disorders (OPMDs). The current study evaluated the rate and the time of malignant transformation in the various OPMDs in a cohort of patients from southern Taiwan. Parameters possibly indicative for malignant transformation of OPMDs, such as epidemiological and etiological factors, and clinical and histopathological features were also described.MethodsWe followed-up 5071 patients with OPMDs—epithelial dysplasia with oral submucous fibrosis, epithelial dysplasia with hyperkeratosis/epithelial hyperplasia, hyperkeratosis/epithelial hyperplasia, oral submucous fibrosis, lichen planus, and verrucous hyperplasia—between 2001 and 2010 for malignant transformation.ResultsTwo hundred nineteen of these 5071 OPMD patients (202 men, 17 women; mean age: 51.25xa0years; range: 30–81 years) developed oral cancers (179 squamous cell carcinomas; 40 verrucous carcinomas) in the same sites as the initial lesions at least 6xa0months after their initial biopsies. The overall transformation rate was 4.32% (mean duration of transformation: 33.56xa0months; range: 6–67 months). Additionally, the mean time of malignant transformation was significantly shorter for lesions with than without epithelial dysplasia. The risk of malignant transformation was 1.89 times higher for epithelially dysplastic than non-dysplastic lesions. The anatomical site of OPMD and the presence of epithelial dysplasia were significantly associated with malignant transformation. The hazard rate ratio was 1.87 times larger for tongue lesions than for buccal lesions.ConclusionPatients with OPMDs require long-term follow up.

Anti-invasion and anti-tumor growth effect of doxycycline treatment for human oral squamous-cell carcinoma--in vitro and in vivo studies.

Regional lymph node and distant organ metastasis of oral squamous-cell carcinoma (OSCC) has been associated with increased production of matrix metalloproteases (MMPs), and scientific data showed that doxycycline (Dox) could down-regulate the expression of MMPs. The objective of this study was to evaluate the effect of Dox on the expression of MMPs in vitro using the SCC-15 cell line and in vivo SCC-15 xenografted nude mice. SCC-15 cells maintained under distinct culture conditions expressed high levels of pro-MMP-2 and pro-MMP-9; however, as determined by zymography and Western blot analysis, Dox significantly reduced the production of pro-MMP-2 and pro-MMP-9 after 24h of treatment in a dose-dependent manner (2.5-40 microg/ml). Dox (10 microg/ml) decreased the expression of MMP-9 mRNA but did not alter the level of MMP-2 mRNA after 24h of treatment. In addition, this drug significantly inhibited the invasive and migration activities of SCC-15 cells in vitro (>75% inhibition at 10 microg/ml). On the other hand, daily administration of Dox (3mg/mice) restrained tumor growth in SCC-15 xenografted nude mice, with an inhibition rate of 85.6%. Compared with the control group (treated with normal saline), MMP-9 mRNA levels in the fresh tumor tissue decreased upon Dox treatment (P<0.01) while MMP-2 mRNA levels were unchanged. In conclusion, reduced expression of MMP-9 at the transcriptional level and MMP-2 at the post-transcriptional level caused by Dox was found to be associated with decreased invasion of oral SCC in vitro. Moreover, Dox exerted a significant suppressive effect on tumor growth in an in vivo nude mice model. Taken together, these results, to our knowledge, may first imply that Doxycycline has an adjuvant therapeutic effect on OSCC that is associated with inhibition of MMPs expression.

Overexpression of Smad proteins, especially Smad7, in oral epithelial dysplasias

ObjectiveTransforming growth factor β, via membrane-bound receptors and downstream Smad2–4, 7, can modulate tumorigenesis. Smad2 and Smad3 heterodimerize with Smad4, and the complex migrates to the nucleus to regulate the expression of target genes. Smad7 is a key negative regulator of this signaling pathway. This study aimed to examine Smad2–4, 7 expression and phosphorylated Smad2–3 (p-Smad2–3) in oral epithelial dysplasia and compared it with normal oral mucosa, hyperkeratosis/epithelial hyperplasia and squamous cell carcinoma (SCC).Materials and methodsImmunohistochemical staining of Smad2–4, 7 and p-Smad2–3, was performed for 75 samples of human oral mucosa, including hyperkeratosis/epithelial hyperplasia (nu2009=u200920), mild epithelial dysplasia (nu2009=u200911), moderate to severe epithelial dysplasia (nu2009=u200911), and SCC (nu2009=u200943). Normal buccal mucosa samples (nu2009=u20099) were also included.ResultsA significant increase in Smad7 expression was observed in the ascending order of samples of normal oral mucosa, hyperkeratosis/epithelial hyperplasia/mild oral epithelial dysplasia, moderate to severe oral epithelial dysplasia, and well-differentiated oral SCC/moderately to poorly differentiated oral SCC. Additionally, significant increases in Smad7 expression were noted as compared with expression of Smad2–4 and p-Smad2–3 in lesions of hyperkeratosis/epithelial hyperplasia, mild oral epithelial dysplasia, moderate to severe oral epithelial dysplasia, well-differentiated oral SCC, and moderately to poorly differentiated oral SCC.ConclusionsOur results indicate that Smad proteins, particularly Smad7, in oral epithelial dysplasia and SCC could contribute to the attenuation of Smads anti-proliferative signaling in cancer development.Clinical relevanceSmad7 could be a marker for risk of malignant transformation of oral epithelial dysplasia.

Expression of osteonectin/secreted protein acidic and rich in cysteine and matrix metalloproteinases in ameloblastoma.

BACKGROUNDnAmeloblastoma is the most common clinically-significant epithelial odontogenic tumor, and is considered a benign but locally-aggressive tumor of the craniofacial region. Osteonectin/secreted protein acidic and rich in cysteine (SPARC) is induced in response to a number of biological processes such as tumor growth and metastasis, whereas matrix metalloproteinases (MMPs) degrade the extracellular matrix and participate in various biological processes including tumor invasion and metastasis. We hypothesize that SPARC acts with MMPs for the local invasiveness of ameloblastoma. The aim of this study was to examine the association of SPARC with MMP-1, MMP-2, and MMP-9 in ameloblastoma.nnnMETHODnImmunohistochemical expression of SPARC, MMP-1, MMP-2, and MMP-9 as well as co-expression of SPARC and MMP-9 were examined in a cohort of 23 cases of ameloblastoma.nnnRESULTSnSPARC, MMP-1, -2, and -9 were detected in the cytoplasm of the ameloblastic-like columnar cells and stellate-reticulum-like cells as well as in the stromal tissues of fibroblasts and endothelial cells of our cohort of ameloblastoma patients. Furthermore, co-expression of SPARC and MMP-9 were found in 23 cases of ameloblastoma. This may be the first study to demonstrate that the expression level of SPARC was statistically correlated with MMP-9 but not with MMP-1 or -2 in ameloblastoma.nnnCONCLUSIONnOur results suggest a putative association between SPARC and MMPs (especially MMP-9) in ameloblastoma to regulate tumor invasion.

The mRNA expression of placental glutathione S-transferase isoenzyme in hamster buccal-pouch carcinomas using reverse transcription-polymerase chain reaction

Placental glutathione S-transferase (GST-P) may facilitate cell proliferation and inhibit apoptosis, hence allowing for the expansion of a population of initiated tumor cells. The enhanced expression of GST-P at the protein level has been reported previously in chemically induced oral carcinomas in hamster buccal-pouch mucosa but the expression of GST-P at the mRNA level has not yet been demonstrated. The purpose of the present study was to assess the GST-P mRNA expression in 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal-pouch carcinomas using a reverse transcription-polymerase chain reaction (RT-PCR). Thirty-five outbred, young (6 weeks old), male, Syrian golden hamsters (Mesocricatus auratus) were randomly divided into one experimental group (15 animals), and two control groups (10 animals each). Bilateral pouches of a group of 15 animals of the experimental group were painted with a 0.5% DMBA solution three times a week for 12 weeks while each animal of one of the control groups was similarly treated with mineral oil. Another control group of 10 animals was untreated throughout the experiment. Areas of dysplasia and squamous-cell carcinomas with a 100% tumor incidence developed in all of the DMBA-treated buccal pouches. The mineral oil-treated and untreated pouches revealed no obvious changes. Placental glutathione S-transferase mRNA was demonstrated to be present amongst all the 12-week DMBA-treated hamster buccal-pouch mucosa animals, but not for the untreated animals or the animals for which the buccal pouch was treated with mineral oil. Multiple potential regulatory pathways including gene amplification, enhanced mRNA stability, chromosomal translocation/gene rearrangement, and hypomethylation of the promoter region can contribute to the overexpression of GST-P mRNA in DMBA-induced hamster buccal-pouch carcinomas. Further study is necessary to completely understand which candidate mechanism(s) will contribute principally to the increased GST-P mRNA expression in oral experimental carcinogenesis.

Prevalence of ponticuli posticus among patients referred for dental examinations by cone-beam CT.

BACKGROUND CONTEXTnPonticulus posticus (PP) is the bony bridge that can completely or partially embrace the vertebral artery and the suboccipital nerve root at the atlas posterior arch. The PP can be a possible cause of vertigo, vertebrobasilar insufficiency, neck pain, shoulder pain, and cervicogenic headache. Moreover, the vertebral artery injury may happen during atlas lateral mass screw insertion in the presence of PP.nnnPURPOSEnThe purpose of this study was to determine the prevalence of PP in a population of patients undergoing dental cone-beam computed tomography (CBCT) and the association between PP and atlas superior articular facet (SAF).nnnSTUDY DESIGNnThis is a retrospective study.nnnPATIENT SAMPLEnFive hundred consecutive patients who had undergone dental CBCT scans were included.nnnOUTCOME MEASURESnOutcome measures were age, sex, and radiologic measures.nnnMETHODSnThe maximum anteroposterior and transverse dimensions of atlas SAF were measured on the axial image, and then the area was calculated by using the formula for an elliptical area. The left-right differential ratios of the SAF in patients with unilateral PP were compared with those in age- and gender-matched patients without PP. The relationships among imaging findings, age, and sex were assessed with the two-tailed paired t test, χ(2) test, and logistic regression model, as appropriate.nnnRESULTSnThe overall prevalence of PP was 7% (35 of 500 patients). There were no significant differences in the prevalence of PP with gender and age. The anteroposterior dimension, transverse dimension, and area of atlas SAF on the PP side were significantly larger than those on the non-PP side in the 18 unilateral complete PP patients (p<.001, p<.001, and p<.001, respectively) and in the 11 unilateral partial PP patients (p=.001, p=.007, and p<.001, respectively). The SAF area differential ratios in patients with unilateral PP were greater than those in the patients without PP (29.8% vs. 2.9%, p=.002 for 18 complete lesions, and 23.5% vs. 1.8%, p<.001 for 11 partial lesions).nnnCONCLUSIONSnThe prevalence of PP and the measurement of SAF can be assessed by CBCT. The imaging findings show the larger SAF on the PP side and greater left-right difference of SAF area in the patients with unilateral PP.

Aberrant expression in multiple components of the transforming growth factor-β1-induced Smad signaling pathway during 7,12-dimethylbenz[a]anthracene-induced hamster buccal-pouch squamous-cell carcinogenesis.

UNLABELLEDnTransforming growth factor (TGF)-β1 signaling controls a plethora of cellular processes including tumorigenesis. The TGF-β1 ligand initiates signaling by binding to TGF-βreceptor II (TβRII) and allowing heterodimerization with TGF-βreceptor I (TβRI); thus, TβRI is phosphorylated by TβRII. After phosphorylation, Smad2 and Smad3 heterodimerize with Smad4, and this complex migrates to the nucleus to regulate the expression of specific target genes. However, Smad7 interrupts above signal transduction by preventing phosphorylation of Smad2 or Smad3. The objective of this study was to examine the TGF-β1-induced Smad signaling pathway during 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal-pouch squamous-cell carcinogenesis. Fifty 6-week-old male Syrian golden hamsters were divided into three experimental and two control groups (10 animals in each). Both pouches of each animal in the experimental groups were painted with 0.5% DMBA solution, and both pouches of each animal of one of the control groups were similarly treated with mineral oil; the other control group remained untreated throughout the experiment. Animals from three experimental groups were sacrificed at the end of 3rd, 9th, and 14th-weeks after DMBA treatment, respectively, and animals from two control groups were all sacrificed at 14th-weeks after the treatment. Immunohistochemical staining for TGF-β1, TβRI, TβRII, Smad2-4 and Smad7 were performed.nnnRESULTSnA significant increase in the expression of Smad7 and significant decreases in the expression of TβRII, Smad 2, Smad3 and Smad4 were noted during hamster buccal-pouch carcinogenesis induced by DMBA. Our findings indicate that a disruption in TGF-β1-induced Smad signaling occurs as a result of aberrant expression of multiple components in the TGF-β1 signaling pathway during DMBA-induced hamster buccal-pouch carcinogenesis, leading to loss of TGF-β1 growth-suppressive effects on transformed pouch keratinocytes.

Human dental pulp stem cells derived from cryopreserved dental pulp tissues of vital extracted teeth with disease demonstrate hepatic-like differentiation.

Reviewing the literature, hepatic differentiation of human dental pulp stem cells (hDPSCs) from cryopreserved dental pulp tissues of vital extracted teeth with disease has not been studied. This study is aimed to evaluate the hypothesis that hDPSCs from cryopreserved dental pulp tissues of vital extracted teeth with disease could possess potential hepatic differentiation. Forty vital extracted teeth with disease recruited for hDPSCs isolation, stem cell characterization and hepatic differentiation were randomly and equally divided into group A (liquid nitrogen‐stored dental pulp tissues) and group B (freshly derived dental pulp tissues). Samples of hDPSCs isolated from groups A and B but without hepatic growth factors formed negative controls. A well‐differentiated hepatocellular carcinoma cell line was employed as a positive control. All the isolated hDPSCs from groups A and B showed hepatic‐like differentiation with morphological change from a spindle‐shaped to a polygonal shape and normal karyotype. Differentiated hDPSCs and the positive control expressed hepatic metabolic function genes and liver‐specific genes. Glycogen storage of differentiated hDPSCs was noted from day 7 of differentiation‐medium culture. Positive immunofluorescence staining of low‐density lipoprotein and albumin was observed from day 14 of differentiation‐medium culture; urea production in the medium was noted from week 6. No hepatic differentiation was observed for any of the samples of the negative controls. We not only demonstrated the feasibility of hepatic‐like differentiation of hDPSCs from cryopreserved dental pulp tissues of vital extracted teeth with disease but also indicated that the differentiated cells possessed normal karyotype and were functionally close to normal hepatic‐like cells. Copyright

Placental glutathione S-transferase isoenzyme expression during promotion of two-stage hamster cheek-pouch carcinogenesis

Glutathione S-transferases (GSTs) are the products of a multigene family. A well-established function of GSTs is to metabolize carcinogens by catalysing the conjugation of electrophilic substrates to glutathione. Whether placental GST (GST-P) is expressed during the promotion of two-stage hamster buccal-pouch mucosa (HBPM) carcinogenesis was investigated here, using 7,12-dimethylbenz[a]anthracene (DMBA) as the initiator and 12-O-tetradecanoylphorbol-13-acetate (TPA) as the promoter. Cytoplasmic and nuclear staining for GST-P was seen in pouches treated with DMBA for 4 or 16 weeks, as well as in those treated with DMBA for 4 weeks and then TPA for 12 weeks. No GST-P positivity was seen in any pouches treated with only TPA or with mineral oil for either 4 or 16 weeks. The average number of GST-P-stained foci in the groups treated with DMBA for 16 weeks (246 +/- 96; mean +/- SD) or DMBA for 4 weeks followed by TPA for 12 weeks (186 +/- 67) was significantly higher than in pouches treated with only DMBA for 4 weeks (97 +/- 24). These results demonstrate that TPA alone is not sufficient for GST-P expression in hamster buccal pouch mucosa. However, after being initiated with DMBA, then promoted with TPA, GST-P activity is induced in hamster buccal pouch mucosa during squamous-cell carcinogenesis. This underpins the suggestion that GST-P may play an important part during the promotion stage of oral carcinogenesis.