Association of HLA-DRB1, HLA-DQB1 Polymorphisms with HPV 16 E6 Variants among Young Cervical Cancer Patients in China.

Potential correlation of human papillomavirus (HPV) 16 E6 variants and human leukocyte antigen (HLA) class II polymorphisms has been suggested in patients with cervical cancer, so far little information is available about the possible interaction between E6 variants and HLA class II variability during the obviously accelerated progression to cervical cancer in young women. In this study, we aimed to explore the association between the HPV16 E6 variants and HLA-DRB1, DQB1 alleles in a Chinese young cervical cancer population. The HLA-DRB1, HLA-DQB1 polymophisms were genotyped by low-resolution polymerase chain reaction (PCR) with sequence-specific primer. HPV16 E6 DNA was tested by Sanger fluorescent dye dideoxy-termination technique. The difference of DRB1, DQB1 polymorphisms between young cervical cancer patients (≤35ys, n=61) and older ones (>35ys, n=85) and the association with E6 variants were analyzed. Results showed that the distribution pattern of HLA-DRB1, DQB1 alleles was different between young cervical cancer patients and older ones. The allele frequency of DQB1*0501 in young patients was significantly lower than older ones (6.6% vs. 23.5%, p<0.05). The HPV16 E6 A4 lineage was the exclusive type observed in young patients, and its prevalence was significantly higher than that of older cases (82.86% vs.41.94%, p<0.05). DRB1*03 was not found in young patients positive for the HPV16 E6 A4 lineage, whereas it was observed in 19.2 % older patients with A4 positive(Pc<0.05). In conclusion, specific association between certain HPV16 E6 variant and genetic polymorphisms of HLA may play a role during the progression of early onset cervical cancer in young patients. Certain HLA-DRB1 and HLA-DQB1 alleles may be related to the A4 lineage among young cervical cancer patients, which was the unique HPV16 E6 variant found in Chinese young patients. Our finding may provide an insight into the pathogenic factors that associated with cervical cancer in young women.

Human papillomavirus type 16 e6 gene variations in young Chinese women with cervical squamous cell carcinoma.

Human papillomavirus (HPV) 16 E6 gene mutation is considered an important genetic change in cervical lesion progression. To explore the possible association of specific HPV16 E6 sequence variations with the development of invasive cervical squamous cell carcinoma (SCC) in young women, we examined the distribution of HPV16 E6 variants in a Chinese cervical SCC population and analyzed the difference between younger patients (≤35 years, n = 50) and older ones (>35ys, n = 71). Human papillomavirus type 16 E6 DNA was amplified by polymerase chain reaction and sequenced by Sanger fluorescent dye dideoxy-termination method. Analysis revealed that the most frequently found variation in this Chinese population was the EV (As) lineage (65.45%). In addition, the EV (As) lineage seems more common and uniform in younger patients than other lineages, and it may be associated with early age at diagnosis of cervical SCC in young women.

[Polymorphism of HLA-DRB1 and -DQB1 alleles among young women with cervical squamous cell carcinoma and its relation with human papillomavirus type 16 infection].

OBJECTIVE: To investigate the polymorphism of human leukocyte antigen (HLA)-DRB1 and -DQB1 alleles among young women with cervical squamous cell carcinoma (SCC) and elucidate its relationship with human papillomavirus (HPV) type 16 infection. METHODS: From January 2005 to August 2009, 166 women diagnosed with cervical SCC at our hospital were enrolled. These patients were divided into two groups based on age, including 59 cases in young age group (≤ 35 yrs) and 107 cases in non-young age group. In the mean time, 50 cases with uterine myoma treated by hysterectomy were selected as controls. HPV 16 DNA in cervical tissues was amplified by polymerase chain reaction (PCR). HLA-DRB1 and -DQB1 typing were carried out by PCR-sequence specific primer (PCR-SSP) and the allele frequencies calculated. RESULTS: (1) The allele frequency of HLA DQB1*0301 at 29.6% was detected among HPV 16 positive cervical SCC cases in young age group. And it was significantly higher than 12.9% of non-young age group (P < 0.05). The allele frequencies of HLA-DRB1*04 and -DRB1*09 were significantly higher among HPV 16 negative cervical SCC cases in young age group as compared with non-young age group (14.1%, 26.6% vs 5.9%, 10.5%) (P < 0.05). The HLA-DRB1*07 allele was not detected among HPV 16 negative cervical SCC cases in young age group, But 14 cases (9.2%) were detected in non-young age group (P < 0.05). (2) The allele frequencies of HLA-DQB1*0501 at 7.4% and 6.3% respectively were detected among HPV 16 positive and negative cervical SCC cases in the young age group. And they were significantly lower than 25.8% and 20.4% of non-young age group (P < 0.05). CONCLUSION: The distribution patterns of HLA-DRB1 and -DQB1 alleles among young women with cervical SCC are different from those of older ones. And it has something to do with the HPV 16 infection status.

[Expression of survivin, bcl-2 in cervical carcinoma and its association with HPV16/18 infection].

OBJECTIVE: To investigate the expression and the correlation of surviving, bcl-2 and HPV16/18 in cervical carcinoma. METHODS: Hybridization in situ was used to detect the expression of survivin mRNA and HPV16/18 DNA in 74 cases of CIN and 81 cases of cervical carcinoma and 20 cases of normal cervical tissues. And immunohistochemical analysis was used to detect the expression of bcl-2 protein. RESULTS: The positive rates of survivin mRNA, bcl-2 and HPV16/18 in CIN were 44.6%, 39.2% and 41.0% respectively versus 77.8%, 70.4% and 81.2% in cervical carcinoma. The above three indices gradually rose in normal cervical tissue, CIN and cervical carcinoma. The expression of survivin and bcl-2 in CINIII were obviously higher than those in CINI/II. And it was obviously higher in cervical carcinoma with stage IIb-III than those in stage I-IIa. And it was also obviously higher in cervical carcinoma with a poor differentiation than those with a good or medium differentiation. The expression of survivin in cervical carcinoma with lymphatic metastasis was significantly higher than that without lymphatic metastasis. There were no relationship between the expression of survivin or bcl-2 and the pathological type or tumor type of cervical carcinoma. The infection of HPV16/18 also had nothing to do with the clinical stage or pathological type or tumor type of cervical carcinoma. Inverse correlation was both observed in the expression of survivin and bcl-2 with survival rate. Thus a positive correlation between surviving, bcl-2 and HPV 16/18 was observed in cervical carcinoma. CONCLUSION: Survivin, bcl-2 and HPV16/18 participate in the development of cervical carcinoma. It may be a useful guide in early diagnosis of cervical carcinoma, evaluation of surgery and chemotherapy and prediction of outcome.

[Expression of FHIT genes in CIN and cervical carcinoma and the relationship between FHIT gene and p53 and HPV16/18].

OBJECTIVE: To investigate the role and significance of FHIT genes depletion, p53 overexpression and HPV16/18 infection in cervical intraepithelial neoplasia (CIN) and cervical carcinoma (CC). METHODS: Tumor samples taken from 52 cases of CIN and 69 cases of CC were processed by immunohistochemistry (SP) to determine the expression of FHIT genes and p53 protein, by in situ hybridization to detect HPV16/18 infection, and were compared with those in 18 cases of normal cervical tissues as control. RESULTS: (1) The FHIT expression was positive in normal cervical tissue with no depletion occurred, and was 30.8% in CIN. It was significantly higher in CIN III and carcinoma groups than that in normal and CIN I/II groups (P < 0.01). The depleted expression of FHIT in infiltrating cervical carcinoma group was 66.7% (46/69), significantly higher than that in normal and CIN groups (P < 0.01). Along with the decreasing of cell differentiation, the negative rate of FHIT raised. (2) The positive expression of p53 in CC group was 56.5% (39/69) and the HPV16/18 was 84.1% (58/69), both higher than that in CIN and normal groups (P < 0.05). (3) In CIN and CC groups, the positive rate of p53 in cases with positive or negative FHIT expression was similar (P > 0.05). (4) There is a negative correlation between FHIT and p53 expression. The rate of HPV16/18 infection in the depleted expression of FHIT group was significantly higher than that in FIHT normal expression group (P < 0.01). CONCLUSION: (1) The FHIT-depletion is related with cervical carcinogenesis. It may be used as a marker to serve mass screening of CIN-high risk subjects and diagnostic indicator for early cervical carcinoma. (2) Depleted expression of FHIT is frequently associated with p53 over-expression in CIN and CC subjects, but there is no direct correlation between them. (3) HPV16/18 infection may probably be the common cause leading to altered FHIT and p53 expression.

[The application of interphase fluorescence in situ hybridization to the diagnosis of X chromosomal count abnormality in ovarian carcinoma cell].

To study the technique of fluorescence in situ hybridization (FISH) and its application value in the diagnosis of sex chromosomal count abnormality in ovarian carcinoma cell. Biotin labeled alpha satellite X chromosome DNA(pBamX7) probe was hybridized with pre-treated slides of ovarian carcinoma cell interphase nucleus in 18 cases of ovarian carcinoma specimens. The slides were treated with Avidin-FITC and Anti-avidin, amplified with an additional layer and counter-stained with PI in antifade solution. The hybridization signals as well as interphase nucleus settings were observed with WIB filters under fluorescence microscope Olympus AX-70, and the number of interphase nucleus in the ovarian carcinoma cell was counted. It was observed under the microscope that the Biotin labeled pBamX7 probe showed green hybridization signals. Cytoplasm counter-stained with PI showed reddish orange. Increased chrosome X copy number was observed in 11/18(61%) ovarian carcinoma specimens, of which the rest 7 (39%) had no increase of chrosome X copy number. Gain of X chrosome had a certain incidence in ovarian cancers, which played a role in the recurrence and development of ovarian cancers. Its significance needs further investigation.