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ABSTRACT: Deregulated expression of high-risk human papillomavirus oncogenes (E6 and E7) is a pivotal event for pathogenesis and progression in cervical cancer. Both viral oncogenes are therefore regarded as ideal therapeutic targets. Small interfering RNAs (siRNA) or double-stranded RNAs can knock down target genes effectively through siRNA-induced transcriptional gene silencing (TGS). Here, we established lentiviral-vector mediated shRNA (LV-shRNA) targeting common promoter of HPV16 E6/E7 and targeting E6 transcript, transduced the lentiviral construct into cervical HPV16-positive cell lines Siha and Caski, then selected and established stably transduced monoclonal cell lines. The results showed that LV-shRNA targeting promoter, as well as targeting E6 transcript, effectively knocked down E6 and E7 expression, resulted in accumulation of p53 and pRB protein and decrease of MCM7 and p16 protein, and consequently remarkably reduced the abilities of proliferation and invasiveness of cervical cancers cells in vitro. Then we inoculated subcutaneously those monoclonal cells into nude mice to establish the transplanted tumor animal models, and found dramatically inhibited tumorigenesis and growth, as well as prolonged survival time of mice incubated by cells with LV-shRNA targeting promoter and E6 transcript. Our results may provide evidence for application of LV-shRNA targeting HR-HPV key oncogenes, as a new treatment strategy, in cervical and other HPV-associated cancer therapy.
Antiviral research 03/2013; · 3.61 Impact Factor
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ABSTRACT: Angiogenesis is a vital process for proper embryonic development, wound healing, malignant tumor growth and metastasis. Two glia maturation factor genes, glia maturation factor-β (GMFB) and glia maturation factor-γ (GMFG), presenting different expression patterns and distinct biological functions are found in vertebrates. But, the role of GMFB and GMFG in vascular development remains largely unknown. Here, we showed that both GMFB and GMFG are highly conserved in vertebrates. Whole-mount in situ hybridization and quantitative RT-PCR results revealed that GMFB and GMFG were differently expressed during zebrafish embryogenesis. GMFB was highly enriched in brain and GMFG was predominantly expressed in endothelial cells. By gene specific MO, knockdown of GMFG, but not GMFB, severely disrupted angiogenic sprouting of intersegmental vessels (ISVs), but this angiogenic defects were prevented by overexpression of a MO-resistant form of zebrafish GMFG mRNA. In addition, the expressions of angiogenic factors VEGF-A, STAT3, MMP2, MMP9, and MMP13 were significantly decreased in endothelial cells of GMFG morphants. Our findings provide the first in vivo evidence that GMFG is an important regulator for angiogenic sprouting during angiogenesis in zebrafish and suggest that GMFG may act as a novel potential target for anti-angiogenesis therapy in clinical settings.
Experimental Cell Research 01/2013; · 3.58 Impact Factor
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ABSTRACT: The aim of this study was to investigate the role of the cell surface expression levels of HLA class I and II molecules, the costimulatory molecules CD80/B7-1 and CD86/B7-2, and the adhesion molecules CD54 and CD58 during cervical carcinogenesis. The expression levels of MHC class I and II molecules, the costimulatory molecules CD80/B7-1 and CD86/B7-2 and the adhesion molecules CD54 and CD58 on CD14+ peripheral blood monocytes (PBMs) from 21 cases of cervical intraepithelial neoplasia (CIN) II-III, 51 squamous cervical carcinomas (SCCs) and 18 healthy controls were analyzed using flow cytometry analysis. We found increased expression levels of HLA-DR (p=0.000), HLA-DQ (p=0.000), CD86/B7-2 (p=0.002) and CD58 (p=0.000) on PBMs from patients with SCC and CIN II-III, compared with healthy control subjects, whereas no significant difference existed in the expression levels of HLA class I antigens, HLA-DP CD80/B7-1 and CD54. Upregulated expression levels of HLA-DR, HLA-DQ, CD86/B7-2 and CD58 were associated with disease progression, indicating that an increased expression of HLA-DR, HLA-DQ, CD86/B7-2 and CD58 on PBMs may be correlated with the evolution of cervical cancer.
Molecular Medicine Reports 09/2012; 6(6):1301-4. · 0.42 Impact Factor
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ABSTRACT: One of the subunits in the mammalian transcription factor II H complex, XPD (TFIIH p80), plays a significant role in the nucleotide excision repair pathway. Events such as abnormal DNA excision repair may be involved in the cervical carcinogenesis process. Expression of the human XPD protein was examined using immunohistochemistry in 84 normal cervical tissues and 148 cervical squamous cell carcinoma samples. Additionally, qRT-PCR was performed to analyse the XPD mRNA expression in 69 fresh normal cervical tissues and 110 cervical carcinoma samples. The relationships between XPD expression and various clinicopathological features (including age, FIGO stage, tumor size, stroma involvement, lymph node metastasis and histologic grade) were assessed. The XPD (TFIIH p80) protein was detected primarily in the cytoplasm. We found a statistically significant difference in XPD expression level in cervical carcinoma versus normal cervical tissue (Z = -7.302, P = 0.000). Notably, XPD mRNA was significantly over-expressed in cervical carcinoma tissues but not in normal cervix tissues (t = 6.942, P = 0.000). However, no statistically significant relationship was found regarding XPD expression and age, FIGO stage, tumor size, stroma involvement, lymph node metastasis or histologic grade (P = 0.089, 0.953, 0.809, 0.275, 0.421, 0.387 respectively). Our results showed that XPD was highly expressed in cervical squamous cell carcinoma tissues. A poorly understood change may occur during the XPD transcription process, resulting in the abnormal enrichment seen from mRNA to the protein level, thus leaving the protein unable to perform the normal function of excision repair. There is a need for further research in order to elucidate the specific mechanism involved.
Pathology & Oncology Research 07/2012; 18(4):969-75. · 1.37 Impact Factor
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ABSTRACT: Podocalyxin-like protein 1 (PCLP1) may be involved in the invasion and metastasis of tumors. However, to date the role of PCLP1 in the progression of epithelial ovarian carcinoma has not been investigated.
PCLP1 expression was examined by immunohistochemistry in 471 cases with various degrees of ovarian epithelial lesions, including 46 cases of normal ovarian epithelial tissue, 105 benign serous tumors, 74 borderline serous tumors, 94 serous carcinomas, 58 benign mucinous tumors, 50 borderline mucinous tumors and 44 mucinous carcinomas. Associations between PCLP1 expression and various clinical characteristics were analyzed.
PCLP1 expression in mucinous carcinoma and borderline mucinous tumor tissues was found to be significantly lower than that observed in normal and benign mucinous tumor tissue. In addition, PCLP1 expression was significantly lower in mucinous carcinoma patients in advanced clinical stage and with poor differentiation of tumor cells. No positive results were observed in serous carcinomas.
Our findings suggest that PCLP1 may be involved in the progression of ovarian mucinous lesions but not in serous lesions. Low PCLP1 expression may be a potential predictor of a poor prognosis in mucinous carcinomas.
Pathobiology 06/2012; 79(6):307-13. · 1.18 Impact Factor
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ABSTRACT: The aim of the present study was to investigate the correlation between fibrinogen level, platelet count and prognosis in patients with epithelial ovarian cancer (EOC).
Preoperative fibrinogen level and platelet count in 136 EOC patients and 146 patients with benign ovarian tumor, and their associations with clinicopathologic parameters and survival in EOC patients, were retrospectively analyzed.
The fibrinogen level in EOC was higher than that in benign patients (3.95 ± 1.37 g/L versus 2.88 ± 0.6 g/L, P < 0.001), and 36.0% (49/136) of EOC patients had hyperfibrinogenemia (fibrinogen >4.0 g/L). The platelet count in EOC was higher than that in benign patients (251.5 ± 89.4 × 10(9) /L versus 206.7 ± 49.0 × 10(9) /L P < 0.001), and 7.4% (10/136) of EOC patients had thrombocytosis (platelet count >400 × 10(9) /L). Hyperfibrinogenemia was associated with International Federation of Gynecologists and Obstetricians (FIGO) stage, non-optimal cytoreduction and poor chemo-response, but not with histologic type and grade, CA-125 level, chemotherapy method, and age. EOC patients with advanced disease showed higher rate of elevated thrombocyte count than patients with early disease (30.7% versus 8.3%, P = 0.002). The rate of thrombocytosis was higher in patients with hyperfibrinogenemia than in those with normal fibrinogen (9/10 versus 1/10, P < 0.001). A significant correlation between platelet count and fibrinogen level was observed in EOC patients (P < 0.001). In multivariate analysis, overall survival was influenced by tumor stage (P < 0.001), chemotherapy with taxane (P < 0.001) and fibrinogen level (P = 0.004), and disease-free survival was only influenced by tumor stage (P < 0.001).
Our findings suggest that hyperfibrinogenemia may be a predictor for poor chemo-response and have a potential role as independent prognostic factors in EOC patients.
Journal of Obstetrics and Gynaecology Research 03/2012; 38(4):651-7. · 0.94 Impact Factor
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ABSTRACT: PARP-1 is a nuclear enzyme that plays an important role in DNA repair, recombination, proliferation and the genome stability. The PARP-1 Val762Ala polymorphism has been associated with increased risk of developing cancers of the prostate, esophagus and lung. The aim of this study was to determine whether the PARP-1 Val762Ala polymorphism is associated with the risk of cervical carcinoma. MA-PCR was used to genotype the PARP-1 Val762Ala polymorphism in 539 women with cervical carcinoma, 480 women with CIN and 800 controls. The genotyping method was confirmed by the DNA sequencing analysis. The PARP-1 Val762Ala polymorphism was not associated with the risk of CIN. However, women carrying the PARP-1 Ala762Ala genotype were significantly susceptible to cervical carcinoma (OR: 2.70, 95% CI: 1.47-3.70), and the similar results were also found in squamous cell carcinoma (OR: 2.56, 95% CI: 1.47-3.70). In HPV positive population, the PARP-1 Ala762Ala genotype was also associated with increased risk of cervical carcinoma (OR: 5.56, 95% CI: 2.08-14.3). Our results indicate that the PARP-1 Ala762Ala genotype increases the risk of cervical carcinoma.
PLoS ONE 01/2012; 7(5):e37446. · 4.09 Impact Factor
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ABSTRACT: Transcriptional silencing of HPV oncogenes using short interfering RNA (siRNA) blocks E6/E7 expression. Our objective was to estimate the effective value of E6/E7 specific siRNA-induced transcriptional gene silencing as a potential therapeutic strategy for cervical cancer.
In vitro studies were performed by employing two categories of siRNA targeting promoter of E6/E7 gene and E7 transcript, respectively, and inhibitory effect of both siRNAs was further observed in vitro and on xenograft in BALB/c mice that were inoculated with siRNA transfected SiHa cells and parental SiHa cells followed by siRNA intratumoral injection in vivo. Tumor volume and growth curves were assessed. Furthermore, cellular proliferation and apoptosis of inoculated tumors were determined by immunohistochemistry staining and TUNEL assay.
The two most active siRNA sequences specifically knockdown E6/E7 expressions at mRNA level in HPV16 positive Siha cells, increased p53 and decreased p16 expressions at protein level, inhibited cell proliferation, and induced cell apoptosis in vitro. Furthermore, both siRNAs effectively inhibited tumor formation and growth no matter in mice with siRNA transfected cells in vitro or with siRNA intratumoral injection in vivo. TUNEL staining and FCM assay consistently showed that tumor retardation was through induction of cellular apoptosis.
RNAi targeting the promoter of HPV16 E6/E7 acts effectively in vitro and in vivo, especially through intratumoral delivery, and may be a candidate therapeutic strategy for cervical cancer.
Gynecologic Oncology 11/2011; 124(2):296-302. · 3.89 Impact Factor
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ABSTRACT: Pelvic lymph node metastases are regarded as the most important risk factor and a predictor of poor prognosis for patients with cervical cancer. Exploration of metastasis-related molecules is helpful toward improving the prognosis in cervical cancer. To identify the role of miR-375 in metastasis and progression of cervical cancer, we examined the expression of miR-375 in 170 cervical cancer tissues and 68 normal cervical tissues, using stem-loop quantitative PCR, and found that the expression of miR-375 in cervical cancer tissues was significantly decreased by 4.45-fold, compared with 68 normal tissues. A significant correlation existed between miR-375 expression and clinicopathologic parameters, including lymph node metastasis of cervical cancer. Overexpressed miR-375 suppressed cell proliferation, blocked G1-to-S cell-cycle transition, and inhibited cell migration and invasion in human cervical SiHa and CaSki cells. SP1, a potential target gene of miR-375, was inversely correlated with miR-375 expression in cervical cancer tissues. Moreover, SP1 was negatively regulated by miR-375, and knockdown of SP1 by siRNA inhibited cell malignant behaviors. Thus, our findings suggest that down-regulated miR-375 promotes cell malignant behaviors via the target gene SP1 and may consequently contribute to the progression of cervical cancer.
American Journal Of Pathology 09/2011; 179(5):2580-8. · 4.89 Impact Factor
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ABSTRACT: Although aberrant miRNAs expression has been documented, altered miR-100 expression in cervical cancer and precursor tissues and its carcinogenic effect and mechanism remain unexplored. The aim of our study was to investigate the role of miR-100 alteration in cervical carcinogenesis.
The expression of miR-100 was examined by quantitative real-time reverse transcriptase PCR (qRT-PCR) in 125 cervical tissues including normal cervical epithelium, cervical intraepithelial neoplasia (CIN), and cervical cancer, as well as in five cervical cell lines. Through modulating miR-100 expression using miR-100 inhibitor or mimic in vitro, cell growth, cycle and apoptosis were tested separately by MTT or flow cytometry and meanwhile Polo-like kinase1 (PLK1) mRNA and protein expressions were detected by qRT-PCR and immunoblotting. The expression of PLK1 in 125 cervical tissues was also examined by immunohistochemical staining and the correlation between miR-100 and PLK1 expression in the same tissues was analysed. Finally, HPV-16 E6/E7 expression was modulated by gene transfection and subsequently the level of miR-100 was examined by qRT-PCR.
The miR-100 expression showed a significantly and gradually reduced tendency from low-grade CIN, high-grade CIN to cervical cancer tissues and a significant decrease in HPV positive cervical cancer cell lines. The modulation of miR-100 expression remarkably influenced cell proliferation, cycle and apoptosis, as well as the level of PLK1 protein, but not mRNA, in vitro experiments. PLK1 expression was negatively correlated with miR-100 expression in CIN3 and cervical cancer tissues. The modulation of HR-HPV E6/E7 expression did not change miR-100 level.
The reduced miR-100 expression participates in the development of cervical cancer at least partly through loss of inhibition to target gene PLK1, which probably occurs in a relative late phase of carcinogenesis. HR-HPV E6/E7 may not directly regulate miR-100 expression in cervical cells.
European journal of cancer (Oxford, England: 1990) 05/2011; 47(14):2166-74. · 4.12 Impact Factor
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Histopathology 05/2011; 58(6):982-4. · 3.08 Impact Factor
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ABSTRACT: Quantitative real-time RT-PCR (RT-qPCR) is being widely used in microRNA expression research. However, few reports detailed a robust identification and validation strategy for suitable reference genes for normalisation in microRNA RT-qPCR studies. The aim of this study was to identify the most stable reference gene(s) for quantification of microRNA expression analysis in uterine cervical tissues. A microarray was performed on 6 pairs of uterine cervical tissues to identify the candidate reference genes. The stability of candidate reference genes was assessed by RT-qPCR in 23 pairs of uterine cervical tissues. The identified most stable reference genes were further validated in other cohort of 108 clinical uterine cervical samples: (HR-HPV- normal, n=21; HR-HPV+ normal, n=19; cervical intraepithelial neoplasia [CIN], n=47; cancer, n=21), and the effects of normalizers on the relative quantity of target miR-424 were assessed. In the array experiment, miR-26a, miR-23a, miR-200c, let-7a, and miR-1979 were identified as candidate reference genes for subsequent validation. MiR-23a was identified as the most reliable reference gene followed by miR-191. The use of miR-23a and miR-191 to normalize expression data enabled detection of a significant dereg-ulation of miR-424 between normal, CIN and cancer tissue. Our results suggested that miR-23a and miR-191 are the optimal reference microRNAs that can be used for normalization in profiling studies of cervical tissues; miR-23a is a novel microRNA normalizer.
Experimental and Molecular Medicine 04/2011; 43(6):358-66. · 2.48 Impact Factor
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ABSTRACT: Acquired resistance to paclitaxel, including regimens, is one of the most significant reasons for treatment failure and death in patients with ovarian cancer, but the causes of this resistance remain unclear. However, cell cycle regulation is a key mechanism by which most chemotherapeutic agents exert their cytotoxic effects.
We created a paclitaxel-resistant ovarian carcinoma cell line from SKOV3 cell line, and the difference of cell cycle distribution was analyzed using flow cytometry. Analysis of human cell cycle pathway complementary DNA array was performed to identify candidate genes associated with paclitaxel resistance. Gene expression changes were validated at the messenger RNA and protein levels by real-time reverse transcriptase polymerase chain reaction and Western analysis, respectively.
The ratio of Gap0/Gap1 phase in SKOV3-TR30 was significantly lower than that in SKOV3 (54.8% ± 6.3% vs 72.7% ± 7.6%, P = 0.035), and the ratio of G2/M phase in SKOV3-TR30 was significantly higher than that in SKOV3 (24.9% ± 6.0% vs 10.2% ± 3.5%, P = 0.021). Complementary DNA microarray analysis demonstrated enhanced glycogen synthase kinase-3α (GSK-3α) expression in paclitaxel-resistant ovarian carcinoma cells. Real-time reverse transcriptase polymerase chain reaction analysis revealed that the paclitaxel-resistant subline exhibited a 7.0 ± 1.8-fold increase in GSK-3α messenger RNA expression. There was a 3.34 ± 0.47-fold increase of total GSK-3 protein (GSK-3α/β) in SKOV3-TR30 cells validated by Western analysis.
This study demonstrates that enhanced expression of GSK-3 is associated with acquired resistance to paclitaxel in ovarian carcinoma cells. Glycogen synthase kinase-3 overexpression may probably be a significant contributor to chemoresistance.
International Journal of Gynecological Cancer 04/2011; 21(3):439-44. · 1.65 Impact Factor
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ABSTRACT: miRNAs have the potential to act on diverse downstream genes, and miRNA signatures of HPV-infected tissues may provide insight into HPV-related carcinogenesis. We set out to profile miRNA expression in HPV-infected samples and relate this to histological and grade-specific alterations in the spectrum of cervical carcinogenesis in vivo. A total of 31 miRNAs showed significant and continuous expression along with the progression from normal cervical tissue to cancer, and six of them were validated in 133 samples. By bioinformatics analyses, we established a putative HPV-associated miRNA-mRNA regulatory network, showing that miR-29 is the most highly enriched. We also found that YY1 and CDK6 were both positively correlated with E6/E7 RNA expression and targeted by tumour-suppressive miR-29. Evidence of miR-29 involvement in HPV infection was further verified in patient samples and by various experimental approaches. Taken together, our results suggest that HPVs have oncogenic properties at least in part by reshaping the milieu of cellular miRNAs. miR-29 restrains cell cycle progression and induces apoptosis via YY1 and CDK6 promoting malignant transformation induced by HPV, although the abnormality of miR-29 in HPV-infected cells might be regulated in an indirect way.
The Journal of Pathology 02/2011; 224(4):484-95. · 6.32 Impact Factor
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ABSTRACT: To evaluate five screening methods of cervical cancer so as to popularize an effective screening strategy for cervical cancer in Zhejiang province.
A total of 1005 women aged 25 - 65 years old were selected from Lishui where cervical cancer was highly prevalent. And 859 subjects were ultimately enrolled between June 2009 and December 2009. Each subject was subjected to five screening methods, including Pap smear, liquid-based cytology (LBC), human papillomavirus DNA with a second-generation hybridization assay (HC2), visual inspection with acetic acid (VIA) and visual inspection with Lugol's iodine (VILI). CIN (cervical intraepithelial neoplasia) 2+ on biopsy was used as the reference standard for disease positivity. Negative colposcopy was accepted as a negative outcome.
The sensitivity (Se), specificity (Sp), positive predictive value (PPV), negative predictive value (NPV) were 25%, 90%, 26.7% and 98.6% for Pap smear; 81.3%, 97.3%, 35.1% and 99.6% for LBC; 68.9%, 82.8%, 7.1% and 99.3% for VIA; 81.3%, 84.6%, 9.1% and 99.3% for VILI; 87.5%, 77.3% and 6.8% for HPV-DNA test respectively.
LBC is associated with a better profile of sensitivity, specificity and predictive value for five screening methods. It has the potential of optimizing the effectiveness of primary cervical cancer screening. Due to a low cost and an easy operation, VIA screening is an effective method of screening cervical cancer in the underdeveloped areas.
Zhonghua yi xue za zhi 02/2011; 91(5):309-12.
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ABSTRACT: The specific intratype HPV genome variations may be associated with the development of cervical cancer in specific geographic regions and a given population. Human papillomavirus (HPV) 58 and 52 have been found to be relatively prevalent among Asian women including Chinese women. This study aimed to assess the risk of HPV 58 and 52 variants for cervical cancer and its precursors in Chinese women.
A total of 2021cervical samples were collected. After DNA extraction and genotyping, a total of 298 (177 HPV58-single positive and 121 HPV52-single positive) DNA samples were analyzed for E6 and E7 sequence variations by direct sequencing.
A total of 29 new reported variations of HPV 58 and 52 were found. For HPV58, the presence of C632T (T20I) and G760A (G63S) variants in E7 showed a positive trend of the association with the severity of neoplasia (P(trend)<0.05, χ² test for trend).
These findings suggest that C632T (T20I) and G760A (G63S) variants in HPV58 E7 are probably risk factors associated with the development of cervical cancer in Chinese women. The presence of HPV58/52 E6 and E7 variants may be different in Chinese women.
Gynecologic Oncology 12/2010; 119(3):436-43. · 3.89 Impact Factor
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ABSTRACT: to explore the value of P16 in the predication of high-grade cervical intraepithelial (HGCIN) by P16 expression in cervical specimens.
one hundred ninety-two residual ThinPrep samples were collected and detected by HPV DNA test and P16 detected by immunocytochemistry. All women underwent colposcopy and histological examination of biopsy specimen if needed. P16 test, cytology and HR-HPV DNA (HC2) test were compared based on histological examination of colposcopic biopsies.
(1) the expression of P16 showed 16.3% in normal or inflammatory cases, 46.7% in CIN 1, 93.8% in CIN 2, 91.1% in CIN 3 and 100.0% in carcinoma. A positive relation between P16 and the grade of cervical lesions was observed by Spearman analysis (P < 0.05, r = 0.900). (2) By P16 test, the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy for ≥ CIN 2 were 94.1%, 78.5%, 77.7%, 94.4% and 85.4% respectively. By HR-HPV DNA test, the sensitivity, specificity, PPV, NPV and accuracy for ≥ CIN 2 were 95.3%, 56.1%, 63.3%, 93.8% and 73.4% respectively. By cytological test, the sensitivity, specificity, PPV, NPV and accuracy for ≥ CIN 2 were 82.4%, 92.5%, 89.7%, 86.8% and 88.0% respectively. There were significant difference of specificity, PPV and accuracy between P16 and HR-HPV DNA (P < 0.05). And no significant difference of accuracy was found between P16 and cytology (P > 0.05).
the specificity, PPV and accuracy of P16 are significantly higher than those of HR-HPV DNA. Thus P16 test is valuable to diagnose HGCIN in ThinPrep specimens.
Zhonghua yi xue za zhi 11/2010; 90(43):3040-4.
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ABSTRACT: Testing for high-risk (HR) human papillomavirus (HPV) infection, a prerequisite of invasive cervical cancer (ICC), has been proposed for primary cervical cancer screening. However, the low specificity limits its clinical significance. Although the risk factors for ICC have been extensively studied, whether the existence of risk factors is associated with the specificity of HR-HPV testing in primary screening is unknown. Our study aimed to evaluate the effects of triage by risk factors on HR-HPV testing for detecting cervical intraepithelial neoplasia grade 2 or worse in cervical cancer screening.
A hospital-based case-control study was conducted to explore the risk factors of ICC, and the predictors obtained were selected as potential triage factors. In population-based screening, potential triage factors associated with cervical intraepithelial neoplasia grade 2 or worse were selected as triage factors, and the performance of HR-HPV testing with triage factors was evaluated.
Lifetime number of sexual partners was selected as a triage factor. With the triage factor, the specificity of HR-HPV testing increased significantly, both in the strata of normal (P = 0.000) and abnormal cytological examination findings (P = 0.030). While stratified by age, no significant differences were observed for women younger than 30 years, but the specificity and the accuracy of the HR-HPV testing increased significantly (P = 0.000 and P = 0.000, respectively) among women aged 30 years or older.
Triage by risk factors is a potentially simple and feasible method to increase the specificity of HR-HPV testing for cervical cancer screening.
International Journal of Gynecological Cancer 11/2010; 20(8):1374-80. · 1.65 Impact Factor
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ABSTRACT: Human papillomavirus (HPV) type 16 is the major etiological agents of cervical cancer. Recently, the studies have demonstrated that HPV intratypic variations could affect oncogenic potential to cervical cancer development. The objectives of this study were to identify HPV-16 E6 and E7 variants prevalent in Chinese women and to assess the risk of them for invasive cervical carcinoma and cervical intraepithelial neoplasia (CIN).
DNA samples were genotyped by flow-through hybridization (HybriMax) and amplified by using primers specific for E6 and E7. Products were directly sequenced and analyzed using BLAST on PubMed.
A total of 170 cervical samples (33 cases of normal control, 11 of CIN 1, 72 of CIN 2-3, and 54 of invasive cervical carcinoma) were HPV-16-positive and were analyzed for E6 and E7 sequence variation. The results showed that HPV-16 Asian lineage was the most frequently detected (77%) and that the Asian variant presented a significantly higher disease risk for cervical cancer and CIN. In addition, 3 novel variants at E6 (Q20P, H118Q, and Q123K) and 2 at E7 (D75N and T86P) were found. The substitution G368T at E6 leading to a premature stop codon occurred in an isolate of normal control sample.
This study reported the distribution of HPV-16 E6 and E7 gene variations in women from southeastern China, which are different from that showed in previous studies and may be important when developing an effective vaccine for this area.
International Journal of Gynecological Cancer 11/2010; 20(8):1391-8. · 1.65 Impact Factor
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ABSTRACT: For quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR), the most commonly used normalization strategy is to select a stable reference gene. However, no suitable reference genes have been identified in cervical tissues to date. The aim of this study was to identify the most stable gene or a set of genes as reference genes for RT-qPCR analysis in cervical tissues from a panel of 12 candidates (ALAS1, PPIA, GAPDH, HBB, TBP, ACTIN, B2M, MBNL2, PGKL, RPLP0, RPL-4, and EEF1A1). In total, 20 normal and 20 cervical cancer specimens were examined. Gene expression data were analyzed using two different statistical models (geNorm and NormFinder). EEF1A1 was identified as the most stable and reliable reference gene, followed by GAPDH and RPLP0, whereas EEF1A1 and GAPDH were the best two-gene combination by NormFinder. The expression validity of EEF1A1 was further determined in 21 normal, 22 cervical intraepithelial neoplasia (CIN(2-3)), and 18 cancer tissues; no expression differences were found among normal, CIN(2-3), and cancer tissues (P>0.05). Our results suggested that EEF1A1 can be used as a reference gene for normalization in gene profiling studies in clinic cervical samples, and the combination of EEF1A1 and GAPDH could be recommended as a much more reliable normalization strategy.
Analytical Biochemistry 10/2010; 405(2):224-9. · 3.00 Impact Factor
