1.Expression of Yes-associated protein 1 gene and protein in oral squamous cell carcinoma.
Song-ying LI ; Ji-an HU ; Hui-ming WANG
Chinese Medical Journal 2013;126(4):655-658
BACKGROUNDOral squamous cell carcinoma (OSCC) is one of the most common malignancies in the oral and maxillofacial region. Yes-associated protein 1 (YAP1) has been implicated as a bona fide oncogene in solid tumors. We seek to elucidate the role of YAP1 in OSCC tissue.
METHODSWe identified YAP1 gene and protein overexpression in 30 OSCC patients and 10 normal oral mucosa tissues by immunohistochemistry, Western blotting and reverse transcription polymerase chain reaction (RT-PCR).
RESULTSIn the normal oral mucosa by immunohistochemical staining, YAP1 mainly located in both the cytoplasm and nucleus mainly the nuclei of the basal cells. In OSCC, the expression of YAP1 translocated from the nucleus to cytoplasm; YAP1 being mainly located in both the cytoplasm and nucleus of the adjacent mucosa. The expression of YAP1 gradual increased in normal oral mucosa, tumor adjacent mucosa and low grade, middle grade, high grade OSCC tissue by Western blotting. Significant difference was found between the expressions of the normal oral mucosa and OSCC tissue (P < 0.05). The coincidence was detected between the normal oral mucosa and OSCC tissue by RT-PCR (P < 0.05).
CONCLUSIONSYAP1 is involved in the carcinogenesis and development of OSCC. There is a transformation between nucleus and cytoplasm.
Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Blotting, Western ; Carcinoma, Squamous Cell ; genetics ; metabolism ; Humans ; In Vitro Techniques ; Mouth Neoplasms ; genetics ; metabolism ; Phosphoproteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction
2.Expression of FHIT and MDM2 in oral submucous fibrosis and canceration tissues.
Xiaomin YIN ; Chunyan WEN ; Yuling HAN ; Yijun GAO ; Zhangui TANG
Journal of Central South University(Medical Sciences) 2010;35(6):572-575
OBJECTIVE:
To determine the role of fragile histidine triad (FHIT) and MDM2 in carcinogenesis of oral submucous fibrosis (OSF).
METHODS:
The expression of FHIT and MDM2 was examined by immunohistochemical S-P method in 44 OSF cases, 15 canceration tissues of OSF, and 10 normal oral mucosa tissues.
RESULTS:
The expression of FHIT was positive in the normal oral mucosa epithelium. The positive expression of FHIT decreased in the OSF and canceration tissues of the OSF.The rate of FHIT positive expression was significantly lower in canceration tissues of OSF than that of the OSF (P < 0.05). The expression of MDM2 was negative in normal oral mucosa epithelium. The positive expression of MDM2 increased in the OSF and canceration tissues of the OSF, and the rate of MDM2 positive expression was significantly higher in the canceration tissues of OSF than that of the OSF (P < 0.05).
CONCLUSION
The loss of FHIT and over-expression of MDM2 may play an important role in the carcinogenesis of OSF.
Acid Anhydride Hydrolases
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genetics
;
metabolism
;
Female
;
Humans
;
Immunohistochemistry
;
Male
;
Mouth Mucosa
;
metabolism
;
Mouth Neoplasms
;
metabolism
;
Neoplasm Proteins
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genetics
;
metabolism
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Oral Submucous Fibrosis
;
metabolism
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Proto-Oncogene Proteins c-mdm2
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genetics
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metabolism
3.The p53-p21(waf1) pathway and centrosome amplification in oral squamous cell carcinomas.
Yang CAI ; Yong-fa LIU ; Hong YANG ; Hong LU
Chinese Journal of Stomatology 2009;44(6):332-335
OBJECTIVETo elucidate the possible role of p53-p21(waf1) pathway for centrosome amplification in oral squamous cell carcinoma (OSCC).
METHODSFormalin-fixed, paraffin-embedded tissues of 8 cases of normal oral epithelium tissues and 27 cases of OSCC tissues were examined for the expression of p21(waf1) and mutated p53 proteins by flow cytometry and immunohistochemistry, and centrosome status was investigated by indirect immunofluorescence double staining with antibodies to centrosome protein gamma-tubulin and cytokeratin. The correlation between p21(waf1), p53 and centrosome amplification in OSCC was statistically analyzed by SPSS 12.0.
RESULTSAll normal oral epithelium tissues showed normal centrosomes (1-2 centrosomes per cell)in epithelium cells, while 21 out of 27 cases (78%) of OSCC showed the evidence of centrosome amplification characterized by supernumerary centrosomes ( >2 centrosomes per cell) in a fraction of tumor cells. The quantity of p21(waf1) protein was lower in OSCC with centrosome amplification [(0.878 +/- 0.081)] than that in OSCC without centrosome amplification [(0.952 +/- 0.018), t = 3.838, P < 0.01], and negative correlations were found between the quantity of p21(waf1) protein and the degree of centrosome amplification (r = -0.472, P < 0.05), as well as the positive staining of p53 (r = -0.491, P < 0.01).
CONCLUSIONSp53-p21(waf1) pathway might involve in centrosome duplication cycle in OSCC. Down-regulated p21(waf1) protein, via p53 transactivation-dependent mechanism, was likely a contributing factor towards centrosome amplification in OSCC.
Carcinoma, Squamous Cell ; metabolism ; pathology ; Centrosome ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; metabolism ; Humans ; Mouth Neoplasms ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; genetics ; metabolism
4.Expression and effect of heterogeneous nuclear ribonucleoprotein A2/B1 in tongue squamous cell carcinoma.
Journal of Central South University(Medical Sciences) 2023;48(5):633-640
OBJECTIVES:
Tongue squamous cell carcinoma (TSCC) is a common cancer in the oral and maxillofacial region, which seriously endangers people's life and health.Heterogeneous nuclear ribonucleoprotein A2/B1(hnRNP A2/B1) is an RNA-binding protein that regulates the expression of a variety of genes and participates in the occurrence and development of a variety of cancers. This study aims to investigate the role of hnRNP A2/B1 in TSCC progression.
METHODS:
The differential expression of hnRNP A2/B1 in oral squamous cell carcinoma (OSCC) and normal oral mucosa cells and tissues was analyzed based on the gene expression profiles of GSE146483 and GSE85195 in the Gene Expression Omnibus (GEO) database. The correlation between hnRNP A2/B1 expression and disease-free survival of TSCC patients was analyzed based on TSCC related chip of GSE4676. TSCC cancer and paracancerous tissue samples of 30 patients were collected in Hunan Cancer Hospital from July to December 2021. Real-time RT-PCR and Western blotting were used to verify the mRNA and protein expression of hnRNP A2/B1 in TSCC patients'samples, respectively. Human TSCC Tca-8113 cells were transfected with hnRNP A2/B1 empty vector (a sh-NC group), knockdown plasmid (a sh-hnRNP A2/B1 group), empty vector overexpression plasmid (an OE-NC group) and overexpression plasmid (an OE-hnRNP A2/B1 group), respectively. The knockdown or overexpression efficiency of hnRNP A2/B1 was detected by Western blotting. The proliferation activity of Tca-8113 cells was detected by cell counting kit-8 (CCK-8), and the apoptosis rate of Tca-8113 cells was detected by flow cytometry.
RESULTS:
Based on the analysis of OSCC-related chips of GSE146483 and GSE85195 in the GEO database, it was found that hnRNP A2/B1 was differentially expressed in the OSCC and normal oral mucosa cells and tissues (all P<0.01). Meanwhile, the analysis of TSCC related chip GSE4676 confirmed that the expression of hnRNP A2/B1 was negatively correlated with the disease-free survival of TSCC patients (P=0.006). The results of real-time RT-PCR and Western blotting showed that the relative expression levels of hnRNP A2/B1 mRNA and protein in TSCC tissues were significantly up-regulated compared with those in adjacent tissues (all P<0.01). The results of Western blotting showed that the expression level of hnRNP A2/B1 in Tca-8113 cells was significantly inhibited or promoted after knockdown or overexpression of hnRNP A2/B1 (all P<0.01). The results of CCK-8 and flow cytometry showed that inhibition of hnRNP A2/B1 expression in Tca-8113 cells reduced cell proliferation activity (P<0.05) and increased cell apoptic rate (P<0.01). Overexpression of hnRNP A2/B1 in Tca-8113 cells significantly increased cell proliferation (P<0.05) and decreased cell apoptosis (P<0.01).
CONCLUSIONS
HnRNP A2/B1 is a key factor regulating the proliferation and apoptosis of TSCC cells. Inhibition of hnRNP A2/B1 expression can reduce the proliferation activity of TSCC cells and promote the apoptosis of TSCC cells.
Humans
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Carcinoma, Squamous Cell/genetics*
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Sincalide/metabolism*
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Tongue Neoplasms/genetics*
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Mouth Neoplasms
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Heterogeneous-Nuclear Ribonucleoprotein Group A-B/metabolism*
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RNA, Messenger
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Tongue/metabolism*
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Cell Line, Tumor
5.Relative quantification of cytokeratin 19 transcription in oral squamous cell carcinoma tissues by fluorescent quantitive real-time RT-PCR.
Lai-ping ZHONG ; Ji-an HU ; Shi-fang ZHAO ; Ze-feng XU ; Fei-yun PING ; Guan-fu CHEN
Chinese Journal of Stomatology 2006;41(9):553-555
OBJECTIVETo investigate the relative quantification of cytokeratin 19 transcription in oral squamous cell carcinoma tissues by fluorescent quantitive real-time reverse transcription-polymerase chain reaction ((RT-PCR).
METHODSCK19mRNA level was detected by fluorescent quantitive real-time RT-PCR in cancerous and para-cancerous tissues from 31 oral squamous cell carcinoma patients. According to the 2(-DeltaDeltaCt) equation, the relative quantification fold of CK19mRNA level was calculated in cancerous tissues compared with para-cancerous tissues.
RESULTSCK19mRNA levels in cancerous tissues were 2.21 folds higher than those in para-cancerous tissues, and the amplicon was specific. CK19mRNA level in cancerous tissue correlated significantly with pathological differentiation degree, the poorer the differentiation was, the higher the CK19mRNA level became.
CONCLUSIONSFluorescent quantitive real-time RT-PCR is accurate and reliable in the detection of relative quantification of CK19 transcription in oral squamous cell carcinoma tissues.
Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; Female ; Humans ; Keratin-19 ; genetics ; metabolism ; Male ; Middle Aged ; Mouth Neoplasms ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; methods
6.Correlation between the expression of cytokeratin-18-mRNA and invasion and metastasis of oral squamous cell carcinoma.
Ping SHU ; De-An ZHAO ; Da-Peng LU
Chinese Journal of Stomatology 2009;44(5):286-288
OBJECTIVETo investigate the relationship between the expression of cytokeratin (CK)-18 and biological behavior of oral squamous cell carcinoma (OSCC).
METHODSTwenty-three patients with OSCC were investigated for the expression of CK-18-mRNA by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Correlations between clinical stages, pathological differentiation, lymphatic metastasis and the expression of CK-18-mRNA were evaluated. CK-18-mRNA expression of peripheral blood from the 23 patients and 23 healthy people were also examined. During follow-up after operation, the peripheral blood was collected again for the expression of CK-19-mRNA.
RESULTSExpression of CK-18-mRNA was found in 16 patients. The expression of CK-18-mRNA was significantly associated with clinical stages, tumor differentiation and lymphatic metastasis. CK-18-mRNA was positive in 4 of 23 blood specimens before operation, but during follow-up only 1 of 23 patients was still positive in peripheral blood.
CONCLUSIONSCK-18 may provide additional information in forecasting the metastasis of OSCC and serve as a reference in monitoring recurrence.
Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Case-Control Studies ; Female ; Humans ; Keratin-18 ; genetics ; metabolism ; Male ; Middle Aged ; Mouth Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; RNA, Messenger ; genetics
7.Telomerase SiRNA inhibits KB cell growth in human oral squamous cell carcinoma.
Zhi-qiang YANG ; Guang-sheng CHEN ; Jia-kun CHEN ; Zhong-liang WU ; Yi-xiong LEI
Chinese Journal of Stomatology 2004;39(5):376-378
OBJECTIVETo test the telomerase SiRNA on telomerase mRNA and on KB cell growth of oral squamous cell carcinoma.
METHODSWe synthesized 21-nucleotide SiRNA duplexes with symmetric 2-nucleotide 3' overhangs corresponding to the target sequence (2 657 approximately 2 675 nucleotide downstream of the start codon) of telomerase mRNA. Telomerase activity, cell proliferation, cell cycle and apoptosis were measured after transfection.
RESULTSTwenty one-nucleotide small interfering RNA (SiRNA) duplexes specifically suppressed expression of endogenous telomerase mRNA in human oral squamous carcinoma KB cells. This inhibitory effect lasted only for about 48 h after transfection. Telomerase activity reduction corresponded to the mRNA suppression. Cell proliferation decreased by 30% at 48 h after transfection and lasted for 120 h after treatment. This inhibitory effect resulted from the block of G(1) to S transition. Apoptosis was not involved in this process.
CONCLUSIONSSiRNA is a powerful tool for studying gene function and can be used as gene-specific therapeutics.
Apoptosis ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Proliferation ; Humans ; KB Cells ; Mouth Neoplasms ; metabolism ; pathology ; RNA, Messenger ; biosynthesis ; RNA, Small Interfering ; genetics ; Telomerase ; genetics ; metabolism ; Tumor Cells, Cultured
8.The relationship of Fas and Fas L protein expression in oral carcinogenesis.
Xin ZENG ; Qianming CHEN ; Minhai NIE
West China Journal of Stomatology 2003;21(4):259-262
OBJECTIVEThe purpose of the study is to investigate the mechanism of immune escape and the expression of Fas and Fas L in oral premalignant lesions (OPLs) and oral squamous cell carcinomas (OSCCs).
METHODS64 samples, including normal oral mucosa(7), hyperkeratosis(9), premalignant lesions(24) and squamous cell carcinomas(24), were studied. The cells in the test specimens, which demonstrate granular staining, were considered as positive. The expression of Fas and Fas L was evaluated semi-quantitatively as follows: -, no expression; + (mild), < 5% positive cells; ++ (moderate), 6%-25% positive cells; +++ (intense), 26%-50% positive cells; or ++++ (very intense), > 50% positive cells.
RESULTSIn the process of oral carcinogenesis, each stage had Fas expression. The positive staining appeared essentially on cell membrane. Various degrees of Fas expression were seen in the diseased tissues. The number of positively stained cells in the moderately and severely dysplastic tissues appeared higher than that in the normal control (P < 0.05). In the OSCC group, the level of expression of Fas antigen decreased significantly by comparison with the normal controls (P < 0.05). Fas L expression was discovered in each stage of the process of oral carcinogenesis. The positive staining appeared in cytoplasm. In hyperkeratotic tissues and OPLs, the number of Fas L expression cells was significantly higher than that in the normal controls. The number of Fas L expression cells of OSCCs increased by comparison with both normal controls and OPLs.
CONCLUSIONThe results indicate that the expression of Fas and Fas L is involved in oral carcinogenesis and this may be directly related to the mechanism by which the cancer cells evade the host immune assault. Perhaps, Fas/Fas L system may be used as a prognostic biomarker in predicting the behavior of oral premalignant lesions in the near future.
Biomarkers, Tumor ; Carcinoma, Squamous Cell ; pathology ; Cell Transformation, Neoplastic ; metabolism ; Double-Blind Method ; Fas Ligand Protein ; Humans ; Membrane Glycoproteins ; biosynthesis ; genetics ; Mouth Mucosa ; pathology ; Mouth Neoplasms ; metabolism ; pathology ; Precancerous Conditions ; metabolism ; pathology ; Random Allocation ; fas Receptor ; biosynthesis ; genetics
9.Expression of osteopontin mRNA in oral squamous cell carcinoma and normal oral mucosa.
Xiao-zhi LU ; Xue-min YIN ; Yuan-yong FENG ; Li-qiong DUAN ; Wan-tao CHEN ; Chen-ping ZHANG
Journal of Southern Medical University 2008;28(7):1165-1167
OBJECTIVETo investigate osteopontin (OPN) mRNA expression in oral squamous cell carcinoma (OSCC) and normal oral mucosa tissues.
METHODSDifferential OPN gene expression were detected in 30 cancerous tissues and their paired normal tissues using real-time reverse transcription-PCR (real-time RT-PCR), and the data were analyzed statistically.
RESULTSReal-time RT-PCR results demonstrated that the relative expression level of OPN mRNA in the cancerous tissues were significantly higher than that in paired normal samples (4.17-/+0.51 vs 0.97-/+0.12, P<0.001), showing a 4.3-fold up-regulation. In the 30 OSCC specimens, OPN mRNA expression in the OSCC of histological grades I showed a 3.1-fold down-regulation, significantly lower than the expression in grade II/III tumors (2.16-/+0.17 vs 6.80-/+0.72, P<0.05); its expression was significantly lower in early stage than in advanced stage OSCCs (2.34-/+0.17 vs 4.73-/+0.35, P<0.05). In cases of cervical lymph node metastasis, the expression was significantly higher than that in cases without lymphatic metastasis (6.38-/+0.56 vs 2.89-/+0.32, P<0.05).
CONCLUSIONOPN mRNA overexpression may play an important role in OSCC carcinogenesis and can be a potential target for OSCC therapy.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; genetics ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Mouth Mucosa ; metabolism ; pathology ; Mouth Neoplasms ; genetics ; pathology ; Osteopontin ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction
10.Circadian variation of clock gene Per2 and cancer-related clock-controlled genes in buccal mucosa carcinoma of golden hamster at different cancer stages.
Hua YE ; Kai YANG ; Xuemei TAN ; Dan ZHAO ; Xiaoqiang LÜ ; Qingqing WANG
West China Journal of Stomatology 2015;33(5):513-518
OBJECTIVEThis study investigates the circadian variation rules of the clock gene Per2 and clock-controlled genes of vascular endothelial growth factor (VEGF), Ki67, c-Myc, and P53 in different stages of carcinogenesis in buccal mucosa carcinoma and their roles in the development of buccal mucosa carcinoma.
METHODSNinety Syrian golden hamsters were housed under. 12 h light/12 h dark cycles. Dimethylbenzanthracene (DMBA) was used to establish the carcinoma model by smearing the golden hamster buccal mucosa. Before DMBA painting and after 6 and 14 weeks, the hamsters were sacrificed at six time points within a period of 24 h (i.e., 4, 8, 12, 16, 20, and 24 h after light onset), and the normal buccal mucosa, precancerous lesions, and cancer tissues were simultaneously obtained. Hematoxylin and eosin stained sections were prepared to observe the canceration of each tissue. Real time polymerase chain reaction was used to detect the mRNA expression of Per2, VEGF, Ki67, c-Myc, and P53. Cosine analysis was employed to determine the circadian-rhythm variations of Per2, VEGF, Ki67, c-Myc, and P53 mRNA expression in terms of median, amplitude, and acrophase.
RESULTSThe expression of Per2, VEGF, P53, and c-Myc mRNA in three different stages appeared with circadian rhythms (P<0.05), whereas the Ki67 mRNA was expressed with circadian rhythm only in normal and precancerous lesion stages (P<0.05). The midline-estimating statistic of rhythms (MESORs) of Per2 and P53 mRNA were significantly down-regulated with the development of cancer (P<0.05), whereas the MESORs of VEGF, c-Myc, and Ki67 mRNA were up-regulated (P<0.05). The amplitude of P53 mRNA significantly decreased with the development of cancer (P<0.05). Moreover, compared with the normal group, the amplitudes of Per2, VEGF, Ki67, and c-Myc mRNA significantly increased in precancerous lesions and cancer tissue (P<0.05). In precancerous stage, the acrophases of Per2, VEGF, and c-Myc mRNA were earlier than that in the normal group, whereas that of Ki67 and P53 mRNA were delayed.
CONCLUSIONThe circadian-rhythm characteristics of the clock gene Per2 and clock-controlled gene expression of VEGF, Ki67, c-Myc, and P53 mRNA have changed with the occurrence and development of carcinoma.
9,10-Dimethyl-1,2-benzanthracene ; Animals ; Carcinogenesis ; Carcinoma, Squamous Cell ; metabolism ; Circadian Rhythm ; Cricetinae ; Mesocricetus ; Mouth Mucosa ; metabolism ; Mouth Neoplasms ; metabolism ; Neoplasm Staging ; Period Circadian Proteins ; genetics ; metabolism ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A