OBJECTIVETo investigate the differential expressions of nucleolin in invasive cervical squamous cell carcinoma, cervical intraepithelial neoplasms (CIN) and normal cervical epithelial tissues and explore the role of nucleolin in the carcinogenesis and progression of cervical squamous cell carcinoma.

METHODSFifty specimens of invasive cervical squamous cell carcinoma, 65 specimens of CIN, and 60 adjacent normal cervical epithelial tissue specimens were examined immunohistochemically for nucleolin expression. The correlation of nucleolin expression levels with histological grades of invasive cervical squamous cell carcinoma and CIN were analyzed.

RESULTSThe specimens of invasive cervical squamous cell carcinoma showed a significantly higher positivity rate for nucleolin expression than CIN and normal cervical epithelial tissues, and the rate in CIN tissues was significantly higher than that in normal cervical epithelial tissues (P<0.01). The expression level of nucleolin was significantly higher in invasive cervical squamous cell carcinoma than in CIN and normal cervical epithelia tissues, and higher in CIN than in normal cervical epithelia tissues, whose immunostaining scores were 7.6±0.3, 6.1±0.2, and 3.0±0.2, respectively (P<0.01). The mean nucleolin immunostaining score was significantly higher in poorly and moderately differentiated than in highly differentiated cervical squamous cell carcinoma (7.9 vs 7.1, P<0.01), and higher in high grade CIN than in low grade CIN tissues (6.0 vs 4.0, P<0.01).

CONCLUSIONSOverexpression of nucleolin plays an important role during carcinogenesis of cervical squamous cell carcinoma and is positively correlated with tumor progression of CIN and cervical squamous cell carcinoma.

Carcinogenesis ; Carcinoma in Situ ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; Disease Progression ; Female ; Humans ; Phosphoproteins ; metabolism ; RNA-Binding Proteins ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; pathology

BACKGROUNDIt was reported that telomerase expression is closely associated with cellular immortality and cancer. This study was designed to investigate the relationship between telomerase expression and the carcinogenesis of cervical cancer, the possible use of telomerase as a marker of cervical intraepithelial neoplasia (CIN) progression or regression, and the natural history of CIN.

METHODSTelomeric repeat amplification protocol (TRAP) assay was used to measure telomerase activity in cervical scrapings and biopsy samples obtained from 105 cases affected with various cervical conditions, including chronic cervicitis (n = 20), CIN (n = 64, 16 cases of CIN I, 20 cases of CIN II, and 28 cases of CIN III), and invasive squamous cell carcinoma (n = 21).

RESULTSIn exfoliated cell samples, telomerase activity was detected in 5 of 20 (25.0%) cases of cervicitis, 10 of 16 (62.5%) cases of CIN I, 11 of 20 (55.0%) cases of CIN II, 23 of 28 (82.1%) cases of CIN III, and 13 of 21 (61.9%) cases of carcinoma. In cervical biopsy samples, telomerase activity was detected in 6 of 20 (30.0%) cases of cervicitis, 8 of 16 (50.0%) cases of CIN I, 9 of 20 (45.0%) cases of CIN II, 27 of 28 (96.4%) cases of CIN III, and 20 of 21 (95.2%) cases of carcinoma. Telomerase activation was significantly higher in CIN samples than in cervicitis samples. Telomerase activity was detected at similar frequency in samples from cervical scrapings and cervical biopsies.

CONCLUSIONThese results seem to suggest that telomerase expression may be associated with carcinogenesis of the cervix. TRAP assay of cervical scraping samples could be used to monitor and predict the development of CIN in clinical practice.

Adult ; Biomarkers, Tumor ; analysis ; Cervical Intraepithelial Neoplasia ; enzymology ; Disease Progression ; Female ; Humans ; Middle Aged ; Telomerase ; metabolism ; Uterine Cervical Neoplasms ; enzymology ; Uterine Cervicitis ; enzymology

OBJECTIVE: To explore whether casticin (CAS) suppresses stemness in cancer stem-like cells (CSLCs) obtained from human cervical cancer (CCSLCs) and the underlying mechanism. METHODS: Spheres from HeLa and CaSki cells were used as CCSLCs. DNA methyltransferase 1 (DNMT1) activity and mRNA levels, self-renewal capability (Nanog and Sox2), and cancer stem cell markers (CD133 and CD44), were detected by a colorimetric DNMT activity/inhibition assay kit, quantitative real-time reverse transcription-polymerase chain reaction, sphere and colony formation assays, and immunoblot, respectively. Knockdown and overexpression of DNMT1 by transfection with shRNA and cDNA, respectively, were performed to explore the mechanism for action of CAS (0, 10, 30, and 100 nmol/L). RESULTS: DNMT1 activity was increased in CCSLCs compared with HeLa and CaSki cells (P<0.05). In addition, HeLa-derived CCSLCs transfected with DNMT1 shRNA showed reduced sphere and colony formation abilities, and lower CD133, CD44, Nanog and Sox2 protein expressions (P<0.05). Conversely, overexpression of DNMT1 in HeLa cells exhibited the oppositive effects. Furthermore, CAS significantly reduced DNMT1 activity and transcription levels as well as stemness in HeLa-derived CCSLCs (P<0.05). Interestingly, DNMT1 knockdown enhanced the inhibitory effect of CAS on stemness. As expected, DNMT1 overexpression reversed the inhibitory effect of CAS on stemness in HeLa cells. CONCLUSION CAS effectively inhibits stemness in CCSLCs through suppression of DNMT1 activation, suggesting that CAS acts as a promising preventive and therapeutic candidate in cervical cancer.
Female ; Humans ; Cell Line, Tumor ; HeLa Cells ; Neoplastic Stem Cells/metabolism* ; RNA, Small Interfering/metabolism* ; Uterine Cervical Neoplasms/metabolism*

OBJECTIVE[corrected] To investigate cathepsin B (CB) expression in squamous cervical carcinoma and its relationship to the clinical and pathological condition.

METHODSCB expression was detected by immunohistochemistry in 56 cases of human invasive squamous cervical carcinoma (ISCC) tissues, 85 cases of cervical intraepithelial neoplasia (CIN) and 38 cases of normal cervical squamous epithelial tissue. The results were analyzed in relation to the grade of differentiation, depth of invasion and pelvic lymph node metastasis.

RESULTSThe positive rates of CB were 87.5% (49/56), 48.3% (41/85) and 48.3% (41/85) in ISCC, CIN and normal tissue, respectively. CB expression in ISCC had significant differences from that ub the CIN and normal tissues (P<0.01). CB positive rates in the tissues with invasion of less than two thirds of the cervix and over two thirds of the cervix were 83.4% (28/34) and 95.5% (21/22) respectively, showing obvious differences between them (P<0.05). CB-positive rates also showed an obvious difference between the tissues with lymphatic metastasis and those without lymphatic metastasis [97.4% (37/38) vs 66.7% (12/18), P<0.05]. CB expression in ISCC was not related to the grade of differentiation.

CONCLUSIONHigh expression of CB is closely associated with tumor infiltration and lymphatic metastasis of cervical cancer.

Adult ; Aged ; Carcinoma, Squamous Cell ; metabolism ; Cathepsin B ; genetics ; metabolism ; Cervical Intraepithelial Neoplasia ; metabolism ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Uterine Cervical Neoplasms ; metabolism

Adult ; Aged ; Capsid Proteins ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Humans ; Middle Aged ; Oncogene Proteins, Viral ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; pathology ; Uterine Cervicitis ; metabolism ; pathology ; Young Adult

OBJECTIVETo investigate the expression of SOX2 in cervical intraepithelial neoplasia (CIN) and cervical cancer and explore its association with the clinical features.

METHODSSOX2 expressions were examined using immunohistochemical method in 10 normal cervical tissue specimens, 36 cervical intraepithelial neoplasia specimens (including 10 cases of grade I, 12 of grade II, and 14 grade III) and 40 cervical cancer specimens (including 21 cases of stage I and 19 of stage II). The correlation between the immunohistochemical results and the clinical features of the patients was analyzed.

RESULTSSOX2 expression was negative in normal cervical tissues, and was positive in 41.6% of CIN specimens (10.0% in CIN I, 41.7% in CIN II, and 64.3% in CIN III) in 82.5% of cervical cancer specimens (78.2% in stage I and 88.2% in stage II). The patients with cervical cancer had a significantly higher positivity rate of SOX2 than normal control group (P<0.05). The positivity rate of SOX2 increased with the evolution of cervical disease. SOX2 protein expression was significantly correlated with the histological grade and lymph node metastasis (P<0.05), but not with the age or clinical stage of the patients (P<0.05).

CONCLUSIONSOX2 expression may serve as a useful indicator for evaluating metastasis and malignancy of cervical cancer.

Adult ; Aged ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; Female ; Humans ; Middle Aged ; Neoplasm Grading ; Neoplasm Staging ; SOXB1 Transcription Factors ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; pathology

In order to investigate the expression levels of Pin1 mRNA and protein in cervical cancer and its association with Ki67 and their clinical significance, amplification of Pin1 gene was examined by RT-PCR, and the expression of both Pin1 and Ki67 protein was detected by immunohistochemistry in cervical cancer tissues. It was shown that the expression levels of Pin1 were higher in cervical cancer than in normal cervical tissues (P < 0.05). The expression of Pin1 protein was increased progressively along with the disease process from normal cervix to CIN and to cervical cancer (P < 0.05). No significant difference in the Pin1 expression was found between disease stages (FIGO), pathological grades or pelvic lymph node metastasis status (P > 0.05). The expression of Pin1 was significantly higher in adenocarcinoma than in squamous carcinoma of the uterine cervix (P < 0.05). In cervical cancer, the overexpression of Pin1 was positively correlated with that of Ki67 (P < 0.05). These results suggested that the overexpression of Pin1 was closely related with cancer cell proliferation or progression of cervical cancer and contributed to oncogenesis. Pin1 may serve as a potential marker for cervical cancer diagnosis.
Cervical Intraepithelial Neoplasia/metabolism ; Ki-67 Antigen/*biosynthesis ; Ki-67 Antigen/genetics ; Peptidylprolyl Isomerase/*biosynthesis ; Peptidylprolyl Isomerase/genetics ; Tumor Markers, Biological ; Uterine Cervical Neoplasms/*metabolism

OBJECTIVETo examine the expressions of HLA class I antigen and CD8 in various cervical diseases and investigate their association with cervical cancer.

METHODSThe expressions of HLA class I antigen and CD8 in cervical tissues sampled from patients with cervical cancer, cervical intraepithelial neoplasia (CIN), and chronic cervicitis were detected using SP immunohistochemistry. The association of the expressions of HLA class I antigen and CD8 with the clinicopathologic indices of the patients was analyzed.

RESULTSThe positive expression rates of HLA class I antigen in cervical cancer, CIN, and chronic cervicitis were 22.6%, 100.0%, and 100.0%, and the positive expression rates of CD8 were 22.6%, 95.5%, and 100.0%, respectively. The positive rates of HLA class I antigen and CD8 were significantly lower in patients with cervical cancer (P<0.01). Patients with stage I cervical cancer had significantly higher positive rates of HLA class I antigen and CD8 than those with stage II cervical cancer (46.7% vs 0.0%, 46.7% vs 0.0%, both P<0.01). The expressions of HLA class I antigen and CD8 decreased with the progression of the clinicopathological stages, and may even become undetectable. The expressions of HLA class I antigen and CD8 were not related to the differentiation degree of the tumor or lymph node metastasis (P>0.05). A positive correlation was found between HLA class I antigen expression and CD8 expression.

CONCLUSIONThe expressions of HLA class I antigen and CD8 are down-regulated or deleted in CIN and cervical cancer, and they may play important roles in the development and progression of CIN and cervical cancer.

Adult ; Aged ; CD8 Antigens ; immunology ; metabolism ; Cervical Intraepithelial Neoplasia ; immunology ; pathology ; Down-Regulation ; Female ; Histocompatibility Antigens Class I ; immunology ; metabolism ; Humans ; Middle Aged ; Uterine Cervical Neoplasms ; immunology ; pathology

OBJECTIVETo investigate the expression of La protein in cervical cancer tissues and explore its role in the occurrence and progression of cervical cancer.

METHODSThe expression of La protein in cervical cancer and normal cervical tissues was detected by immunohistochemical staining. RNA interference technology was used to silence La protein expression in HeLa cells and the changes in cell proliferation, tumor sphere formation and cell cycles were investigated.

RESULTSThe expression of La protein was significantly higher in cervical cancer tissues than in normal cervical tissues (61% vs 9%, P<0.05). Silencing La protein expression in HeLa cells caused significantly reduced the cell proliferation and lowered the tumor sphere formation rate from the control level of (17.1=1.92)% to (6.3=0.45)% (P<0.05), resulting also in G0/G1 cell cycle arrest and reduced cyclin D1 protein expression.

CONCLUSIONThe RNA binding protein La can promote the development of cervical cancer and may play a critical role in the carcinogenesis and progression of cervical cancer.

Autoantigens ; metabolism ; Cell Cycle Checkpoints ; Cyclin D1 ; metabolism ; Female ; HeLa Cells ; Humans ; RNA Interference ; RNA-Binding Proteins ; metabolism ; Ribonucleoproteins ; metabolism ; Uterine Cervical Neoplasms ; metabolism ; pathology

Objective: To explore the relationship between abnormal expression of fragile histidine triad (FHIT) gene and methyl-CpG-binding protein 2 (MeCP2) as well as their interaction on cervical cancerization. Methods: A total of 73 patients with cervical squamous cell carcinoma (SCC), 113 patients with cervical intraepithelial neoplasia (CIN Ⅰ, n=45; CINⅡ/Ⅲ, n=68) and 60 women with normal cervix (NC) were included in the study. Real time PCR and Western blot were performed to detect the expression levels of mRNA and protein about FHIT and MeCP2, respectively. The methylation status of FHIT gene CpG island was tested by methylation-specifc PCR (MSP). Kruskal-Wallis H test, χ(2) test, trend χ(2) test and Spearman correlation analysis were conducted with software SPSS 20.0. The interaction was evaluated by generalized multifactor dimensionality reduction (GMDR) model. Results: With the deterioration of cervical lesion, the methylation rates of FHIT gene CpG island (χ(2)=18.64, P<0.001; trend χ(2)=18.08, P<0.001) increased gradually, while the expression levels of FHIT mRNA (H=27.32, P<0.001; trend χ(2)=12.65, P<0.001) and protein (H=47.10, P<0.001; trend χ(2)=29.79, P<0.001) decreased gradually. There was a negative correlation between the methylation rates of FHIT gene CpG island and the expression level of FHIT protein (r=-0.226, P<0.001). The levels of MeCP2 mRNA (H=26.19, P<0.001; trend χ(2)=11.81, P=0.001) and protein (H=69.02, P<0.001; trend χ(2)=47.44, P<0.001) increased gradually with the aggravation of cervical lesions. There was a positive correlation between the expression level of MeCP2 protein and the FHIT mRNA Ct ratio (r=0.254, P<0.001). Expression of proteins were negatively correlated between MeCP2 and FHIT (r=-0.213, P=0.001). The results analyzed by GMDR model showed that there were interactions among high MeCP2 protein expression, the CpG island methylation of FHIT and mRNA and protein expression in CINⅡ/Ⅲ group, and among high MeCP2 mRNA and protein expression, the CpG island methylation of FHIT and low mRNA and protein expression in SCC group. Conclusion: High expression of MeCP2 mRNA and protein, the CpG island methylation and low mRNA and protein expression of FHIT could increase the risk of cervical carcinogenesis, and there might be a synergistic effect on cervical carcinogenesis.
Acid Anhydride Hydrolases/metabolism* ; Carcinoma, Squamous Cell/pathology* ; DNA Methylation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Methyl-CpG-Binding Protein 2/metabolism* ; Neoplasm Proteins/metabolism* ; Polymerase Chain Reaction/methods* ; RNA, Messenger ; Uterine Cervical Neoplasms/pathology* ; Uterine Cervical Dysplasia/pathology*