The pathogenesis of endocervical glandular lesions are not clearly understood. The aims of this study are to evaluate the etiologic role of human papillomavirus (HPV) 16/18 and the relationship of HPV 16/18, p53 and MIB-1 expressions in endocervical glandular dysplasia (EGD), adenocarcinoma in situ (AIS) and adenocarcinoma. The materials included 14 endocervical adenocarcinoma and 5 AIS and 18 high grade EGD and 39 low grade EGD. Immunohistochemistry for p53 and MIB-1, and in situ PCR for HPV 16/18 were done. HPV 16/18 positivity was 84.2%, 16.7% and 17.9% in malignant glandular lesion (adenocarcinoma and AIS), high grade EGD and low grade EGD, respectively. P53 protein expression rates of malignant glandular lesions, high grade EGD and low grade EGD were 31.6%, 11.1%, and 0%, respectively. High MIB-1 labelling index was found in 73.7% of malignant glandular lesions, but in only 5.7% and 3.6% of high and low grade EGD, respectively. There were statistically significant differences in HPV 16/18, p53 and MIB-1 expressions between malignant endocervical glandular lesions and EGD, but no significant difference in p53 and MIB-1 expressions in relation to HPV 16/18 expression. In malignant endocervical glandular lesions, HPV 16/18 infection may be a major causative factor, but not be related to p53 and MIB-1 expressions.
Adenocarcinoma/pathology/physiopathology/*virology ; Carcinoma, Squamous Cell/pathology/physiopathology/virology ; Cervix Neoplasms/pathology/physiopathology/*virology ; Female ; Human ; Nuclear Proteins/analysis/*genetics ; Oncogene Proteins, Viral/genetics ; *Papillomavirus, Human ; Papovaviridae Infections/*pathology/physiopathology ; Protein p53/analysis/*genetics ; Tumor Virus Infections/*pathology/physiopathology

OBJECTIVETo detect the high-risk human papillomavirus (HPV) infectious condition in women with abnormal cytology and evaluate its values in the screening of high grade squamous intraepithelial lesion.

METHODSWe used hybrid capture 2 (hc2) method to examine 949 patients with abnormal cervical cytology results [ > or =atypical squamous cells of undetermined significance (ASC-US) according to the 2001 The Bethesda System diagnosis criteria]. All subjects also received colposcopy for tissue studies.

RESULTSAmong 949 patients with abnormal cytology, the diagnoses of atypical squamous cells (ASC), low grade squamous intraepithelial lesion (LSIL), and high grade squamous intraepithelial lesion (HSIL) were made in 432, 310, and 207 patients, respectively. The high-risk HPV positive rate in ASC, LSIL, and HSIL were 40.3%, 44.8%, and 89.4%, respectively. The numbers of patients with pathologically confirmed results of negative intraepithelial lesion or malignancy (NILM), cervical intraepithelial neoplasia 1, 2, 3 (CIN 1, 2, 3), and squamous cell carcinoma (SCC) were 335, 388, 118, 101, and 7, and the high-risk HPV positive rate was 17.3%, 66.2%, 92.4%, 97.0%, and 100%, respectively. Among patients with atypical squamous cells of undetermined significance (ASC-US), rate of HSIL in high-risk HPV positive group and negative group were 10.2% and 0.8%, respectively (P < 0.01). In screening HSIL, the sensitivities of cytology [ > or = ASC cannot exclude HSIL (ASC-H)] and cytology ( > or = ASC-H) plus high-risk HPV testing were 0.925 and 0.991, and the specificities were 0.510 and 0.748, respectively (P < 0.01). Sensitivitives of cytology ( > or = LSIL) and cytology (> or = LSIL) plus high risk HPV in detecting HSIL were 0.898 and 0.982, respectively, while the specificitives were 0. 567 and 0.779, respectively (P < 0.01).

CONCLUSIONSThe positive rate of high-risk HPV increases with the gravity of cervical lesions. In patients with abnormal cervical cytology, high-risk HPV testing can improve the sensitivity and specificity in the screening of HSIL.

Carcinoma, Squamous Cell ; diagnosis ; pathology ; virology ; Cervical Intraepithelial Neoplasia ; diagnosis ; pathology ; virology ; Female ; Humans ; Papillomaviridae ; genetics ; isolation & purification ; Papillomavirus Infections ; diagnosis ; pathology ; virology ; Risk Assessment ; Uterine Cervical Neoplasms ; diagnosis ; pathology ; virology

OBJECTIVETo analyze the infection of human papillomavirus in laryngeal squamous cell carcinoma patients.

METHODSThe pathological samples of 64 clinical diagnosed laryngeal squamous cell carcinoma patients were collected. Lunimex and PCR techniques were used to detect the HPV gene infection and immunohistochemistry method was used to analyze the HPV protein expression in the samples.

RESULTSIn the 64 cases, 7 were positive for HPV infection by Luminex and PCR tests. 18 were positive for HPV16/18 E6 protein expression. The total positive rate was about 39. 1%.

CONCLUSIONThe high HPV infection rate in laryngeal squamous cell carcinoma patients in the study indicated indirectly that the importance of the HPV infection in pathogenesis of the laryngeal squamous cell carcinoma.

Carcinoma, Squamous Cell ; pathology ; virology ; DNA, Viral ; genetics ; Female ; Human papillomavirus 16 ; genetics ; Human papillomavirus 18 ; genetics ; Humans ; Laryngeal Neoplasms ; pathology ; virology ; Male ; Middle Aged ; Papillomavirus Infections ; pathology ; virology

OBJECTIVETo transform HPV E6/E7 immortalized human oral epithelial cell (HIOEC) line cells by benzo(a)pyrene [B(a)P] in vitro, and to establish a carcinogenesis model of oral squamous cell carcinoma.

METHODSHIOEC cells were treated with 0.1 mg/L-1.2 mg/L B(a)P for 6 months. The cells were cloned at 18th passage, and then the culture medium was changed into DMEM containing 10% FBS at 21th passage. Cells were cultured in vitro for half and one year and the cell line was named HIOEC-B(a)P. The morphological changes of the cells were observed with differential interference contrast microscope and HE staining. The soft agar colony forming ability and tumorigenicity of the cells in nude mice were identified to confirm the malignant characteristics of HIOEC-B(a)P cells.

RESULTS(1) After HIOEC cells were treated with B(a)P for 6 months, HIOEC-B(a)P cells could grow well in DMEM medium containing 10% FBS and physical concentration of calcium. (2) When HIOEC cells were treated with chemical carcinogens, the morphology of the cells was changed. Cells showed the character of polygon epithelial cells with much atypical mitosis. (3) The 93th passage of HIOEC-B(a)P cells had soft agar colony formation ability. (4) The 55th passage of HIOEC-B(a)P cells could develop parakeratosis mass. The 69th passage of HIOEC-B(a)P cells could develop typical well-differentiated squamous cell carcinoma. The 74th and the 96th HIOEC-B(a)P cells developed I-II grade squamous cell carcinoma-like clinical lesions in nude mice.

CONCLUSIONSB(a)P may induce HIOEC cells to be oral squamous cell carcinoma (OSCC) carcinogenetic cells. It will provide a multiple factors, multistage carcinogenesis model of OSCC for the further research.

Animals ; Benzo(a)pyrene ; toxicity ; Carcinoma, Squamous Cell ; chemically induced ; pathology ; virology ; Cell Line, Transformed ; Cell Line, Tumor ; Cell Transformation, Viral ; Epithelial Cells ; pathology ; Human papillomavirus 16 ; genetics ; Humans ; Mice ; Mice, Nude ; Mouth Neoplasms ; chemically induced ; pathology ; virology ; Neoplasms, Experimental

OBJECTIVETo study the difference in gene expression between human papillomavirus (HPV)16-positive and HPV-negative esophageal squamous cell carcinoma(ESCC) .

METHODSEight HPV 16-positive and seven HPV-negative ESCC specimens were evaluated by PCR. The samples were then determined for gene expression profiling using Solexa Sequencing Chip followed by bioinformatics analysis.

RESULTSA total of 796 differentially expressed genes between HPV 16-positive and HPV-negative ESCC were observed. Among them, 366 were up-regulated while 430 were down-regulated. Functional classification and pathway analysis showed that the functions of these genes were mostly related to tumor morphology, immune, and inflammatory response, cellular growth and proliferation and cellular movement. Of these, factors related to immune and inflammation were the most representative.

CONCLUSIONDifferences in immunologic factors may be associated with HPV infection in esophageal cancer.

Adult ; Aged ; Carcinoma, Squamous Cell ; genetics ; pathology ; virology ; Esophageal Neoplasms ; genetics ; pathology ; virology ; Female ; Gene Expression Profiling ; Human papillomavirus 16 ; genetics ; Humans ; Male ; Microarray Analysis ; Middle Aged ; Papillomaviridae ; genetics ; Papillomavirus Infections ; genetics

OBJECTIVETo investigate 30 bp deletion of latent membrane protein (LMP)-1 gene in lymphoepithelial carcinoma (LEC) of the salivary glands and to determine the frequency of this deletion.

METHODSForty-six cases of salivary gland LEC were investigated by PCR to explore the site specific, 30 bp deletion in the 3' terminal region of LMP-1 gene. To guarantee amplifiable DNA extracted from paraffin-embedded tissue sections, PCR amplification of a house-keeping gene (beta-actin) was performed simultaneously. In addition, DNA sequencing of the PCR product was performed in representative cases.

RESULTSAlthough amplifiable DNA was obtained in 42 of the 46 specimens, as indicated by beta-actin gene amplification, successful amplification of LMP-1 gene was achieved in 35/42 (83.3%) cases. Two types of PCR products of LMP-1 gene were observed and confirmed by DNA sequencing. A wild-type PCR product (316 bp) was present in 31 cases (88.6%) and only 4 cases (11.4%) showed an aberrant 286 bp PCR product, corresponding to the 3' terminal 30 bp deletion of the gene.

CONCLUSIONSite-specific 30 bp deletion of LMP-1 gene is not a common feature of salivary gland LEC.

Base Sequence ; Carcinoma ; genetics ; pathology ; virology ; Carcinoma, Squamous Cell ; genetics ; pathology ; virology ; Epstein-Barr Virus Infections ; epidemiology ; virology ; Gene Deletion ; Herpesvirus 4, Human ; genetics ; Humans ; Molecular Sequence Data ; Oncogene Proteins, Viral ; genetics ; Salivary Gland Neoplasms ; genetics ; pathology ; virology ; Sequence Deletion ; Viral Matrix Proteins ; genetics

OBJECTIVETo define a correlation between human papillomavirus (HPV) type 16 and telomerase activity during the carcinogenesis of oral mucosa.

METHODSHPV16 DNA and human telomerase reverse transcriptase (hTRT) mRNA were detected in 82 cases of paraffin embedded tissues including 7 cases of normal oral mucosa, 7 cases of hyperplasia lesions, 30 cases of oral dysplasia lesions and 38 cases of oral squamous cell carcinomas (OSCCs) by PCR and in situ hybridization (ISH) respectively.

RESULTSHPV16DNA was positive in 14.3% (1/7) of normal oral mucosa, 42.9% (3/7) of hyperplasia lesions, 66.6% (20/30) of dysplasia lesions and 92.1% (35/38) of OSCCs. hTRTmRNA was detectable in 30.0% (9/30) of oral dysplasia lesions and 81.6% (31/35) of OSCCs while normal oral mucosa and hyperplasia lesions were negative. Expression of HPV16DNA and hTRTmRNA were co-ordinate in 67.0% (55/82) cases.

CONCLUSIONSHPV16 infection may play an important role in carcinogenesis of oral mucosa by activation of telomerase.

Carcinoma, Squamous Cell ; enzymology ; virology ; DNA, Viral ; analysis ; DNA-Binding Proteins ; Humans ; Hyperplasia ; Mouth Mucosa ; enzymology ; pathology ; virology ; Mouth Neoplasms ; enzymology ; virology ; Papillomaviridae ; genetics ; isolation & purification ; RNA, Messenger ; analysis ; Telomerase ; genetics

Carcinoma, Squamous Cell ; genetics ; pathology ; virology ; Cervical Intraepithelial Neoplasia ; genetics ; pathology ; virology ; DNA, Viral ; metabolism ; Female ; Gene Amplification ; Humans ; In Situ Hybridization, Fluorescence ; Lymphatic Metastasis ; Neoplasm Grading ; Neoplasm Staging ; Papillomaviridae ; genetics ; Papillomavirus Infections ; metabolism ; RNA ; genetics ; Telomerase ; genetics ; Uterine Cervical Neoplasms ; genetics ; pathology ; virology

Methylation of p16 is an important mechanism in cervical carcinogenesis. However, the relationship between cervical squamous cell carcinoma (SCC) and Epstein-Barr virus (EBV) remains controversial. Here, we explored whether EBV infection and/or p16 gene inactivation would play any role in cervical carcinogenesis. Eighty-two specimens included 41 invasive SCCs, 30 cervical intraepithelial neoplasm (CIN; CIN 1, 11 cases, CIN II, 3 cases, CIN III 16 cases) and 11 nonneoplastic cervices. EBV was detected by polymerase chain reaction (PCR) for EBNA-1 and in situ hybridization for EBER-1. The p16 methylation-status and the expression of p16 protein were studied by methylation-specific PCR and immunohistochemistry, respectively. The materials were divided into four groups: 1) nonneoplastic cervices, 2) CIN I, 3) CIN II-III and 4) invasive SCCs. p16 methylation and p16 immunoexpressions increased in CIN and invasive SCCs than nonneoplastic tissue. p16-methylation and p16-immunoreactivities were higher in the EBV-positive group (p=0.009, p<0.001) than in the EBV-negative group. EBV was detected more frequently in CIN and SCCs than nonneoplastic cervices. In conclusion, a correlation between p16 methylation, p16 immunoreactivity and the detection of EBV strongly suggested that the cooperation of EBV and p16 gene may play a synergic effect on cell cycle deregulation.
Carcinoma, Squamous Cell/genetics/*pathology/virology ; Comparative Study ; Cyclin-Dependent Kinase Inhibitor p16/analysis/*genetics ; *DNA Methylation ; DNA, Viral/genetics/isolation & purification ; Epstein-Barr Virus Infections/genetics/*pathology/virology ; Epstein-Barr Virus Nuclear Antigens/genetics ; Female ; Herpesvirus 4, Human/genetics ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Polymerase Chain Reaction ; Precancerous Conditions/genetics/*pathology/virology ; RNA, Viral/genetics ; Research Support, Non-U.S. Gov't ; Uterine Cervical Neoplasms/genetics/*pathology/virology

OBJECTIVETo study the feasibility of diagnosing of human papillomavirus (HPV) infection in paraffin-embedded cervical tissues by high-throughput gene chip technology and its clinical significance.

METHODSForty cases of HPV-related cervical lesions, including 18 cases of invasive squamous cell carcinoma, 12 cases of cervical intraepithelial neoplasia (CIN) III, 6 cases of CIN II and 4 cases of CIN I, were enrolled. DNA was extracted from paraffin-embedded tissues and amplified by polymerase chain reaction (PCR) using HPV DNA primers. The PCR products were then reversely hybridized with gene chip technology. The results were compared with that of in-situ hybridization (ISH).

RESULTSAll of the 18 cases of cervical squamous cell carcinoma were positive for high-risk HPV genotypes (with 1 case showing a mixture with low-risk genotypes). In contrast, 11 cases (91.7%) of CIN III, 5 cases (83%) of CIN II and none of the CIN I cases were positive for high-risk HPV genotypes. On the other hand, low-risk HPV genotypes were detected only in 1 case (17%) of CIN II and 2 cases (50%) of CIN I. The difference between the two groups (CIN III/squamous cell carcinoma versus CIN I/CIN II) was statistically significant (U = 80.0, P < 0.01). Among the 10 squamous carcinoma cases positive for HPV types 16 and 18 by gene chip technology, high-risk HPV DNA was also detected in 6 of them when using in-situ hybridization.

CONCLUSIONSGene chip technology is able to detect multiple HPV genotypes in paraffin-embedded tissues with high sensitivity and specificity. The distinction between low and high-risk HPV genotypes is seemed useful in prevention and management of cervical cancer.

Alphapapillomavirus ; genetics ; Carcinoma, Squamous Cell ; virology ; Cervical Intraepithelial Neoplasia ; diagnosis ; virology ; Cervix Uteri ; pathology ; virology ; DNA, Viral ; analysis ; Female ; Genotype ; Human papillomavirus 16 ; genetics ; Human papillomavirus 18 ; genetics ; Humans ; Oligonucleotide Array Sequence Analysis ; methods ; Papillomavirus Infections ; diagnosis ; virology ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; Uterine Cervical Neoplasms ; diagnosis ; virology