Characteristics of HPV integration in cervical adenocarcinoma and squamous carcinoma.

PURPOSE: HPV integration usually occurs in HPV-related cancer, and is the main cause of cancer. But the carcinogenic mechanism of HPV integration is unclear. The study aims to provide a theoretical basis for understanding the pathogenesis of cervical adenocarcinoma (AC) and cervical squamous carcinoma (SCC). METHODS: We used HPV capture sequencing to obtain HPV integration sites in AC and SCC, and analyzed cytobands, distribution of genetic and genomic elements, identified integration hotspot genes, clinicopathological parameters, breakpoints of HPV16 and performed pathway analysis. Then we conducted immunohistochemical (IHC) assay to preliminarily verify the expression of most frequently integrated genes in AC, STARD3 and ERBB2. RESULTS: The results revealed that the most frequently observed integrated cytoband was 17q12 in AC and 21p11.2 in SCC, respectively. The breakpoints in both AC and SCC were more tended to occur within gene regions, compared to intergenetic regions. Compared to SCC samples, AC samples had a higher prevalence of genomic elements. In AC, HPV integration has no significantly difference with clinicopathological parameters, but in SCC integration correlated with differentiation (P < 0.05). Breakpoints of HPV in SCC located in LCR more frequently compared to AC, which destroyed the activation of promoter p97. Hotspot genes of HPV integration were STARD3 and ERBB2 in AC, and RNA45S rDNA and MIR3648-1 in SCC, respectively. Meanwhile, we preliminarily proved that the expression of STARD3 and ERBB2, the most frequently integrated genes, would increase after integration. CONCLUSION: These results suggested that HPV may utilize the powerful hosts' promoters to express viral oncogenes and overexpression of viral oncogenes plays a significant role in the carcinogenesis of SCC. In AC, HPV integration may affect hosts' oncogenes, and the dysregulation of oncogenes may primarily contribute to progression of AC.

APTw combined with mDixon-Quant imaging to distinguish the differentiation degree of cervical squamous carcinoma.

Background: To investigate the value of amide proton transfer weighted (APTw) imaging combined with modified Dixon fat quantification (mDixon-Quant) imaging in determining the degree of differentiation of cervical squamous carcinoma (CSC) against histopathologic. Methods: Magnetic resonance imaging (MRI) data were collected from 52 CSC patients. According to histopathologic results, patients were divided into the poorly differentiated group (37 cases) and the well/moderately differentiated group (15 cases). The APTw value by APTw imaging and the fat fraction (FF) and transverse relaxation rate R 2 * values by mDixon-Quant were independently measured by two radiologists. Intra-class correlation coefficients (ICCs) were used to test the consistency of APTw, FF, and R 2 * values measured by the two observers. The Mann-Whitney U test was used to analyze the difference in each parameter between the two groups. Logistic regression analysis was used to assess the association between the degree of differentiation on histopathology and imaging parameters by APTw and mDixon Quant. The ROC curve was used to evaluate the diagnostic efficacy of various parameters and their combination in distinguishing the degree of CSC differentiation on histopathology. The DeLong test was used to access the differences among the area under the ROC curves (AUCs). The Pearson correlation coefficient was used to evaluate the correlation between APTw and mDixon-Quant imaging parameters. Results: The APTw means were 2.95 ± 0.78% and 2.05 (1.85, 2.65)% in the poorly and well/moderately differentiated groups, respectively. The R 2 * values were 26.62 (21.99, 33.31)/s and 22.93 ± 6.09/s in the poorly and well/moderately differentiated groups, respectively (P < 0.05). The AUCs of APTw, R 2 * , and their combination were 0.762, 0.686, and 0.843, respectively. The Delong test suggested statistical significance between R 2 * and the combination of APTw and R 2 * . R 2 * values showed a significant correlation with APTw values in the poorly differentiated group. Conclusions: APTw combined with mDixon-Quant can be used to efficiently distinguish the differention degrees of CSC diagnosed on histopathology.

Comprehensive profile and contrastive analysis of circular RNA expression in cervical squamous carcinoma and adenocarcinoma.

Background: Numerous studies have shown circular RNA (circRNA) dysregulation is associated with the pathogenesis of cervical cancer,particularly in individual carcinoma variants. The aim of this study is to investigate and contrastively analyze the expression pattern of circRNAs in cervical squamous carcinoma and adenocarcinoma mediated by human papillomavirus type 16 (HPV-16). Methods: The expression of circRNAs in cervical squamous carcinoma (SCC), adenocarcinoma (ADC) and adenosquamous carcinoma (ASC) tissues, together with the adjacent normal tissues (ANT), was profiled by high-throughput RNA sequencing (RNA-seq). Bioinformatics analysis and quantitative real time polymerase chain reaction (qRT-PCR) validation of the sequencing data were performed. A network of circRNA-miRNA (microRNA)-mRNA was then constructed according to predicted targets and function of candidate circRNAs. Results: A total of 11,685 annotated circRNAs were identified in six cervical samples. There were 42 up-regulated and 98 down-regulated circRNAs. 215 circRNAs were up-regulated in SCC but down-regulated circRNAs in ADC, while 50 circRNAs displayed the opposite trend. Function enrichment analysis based on different expressions of circRNAs found that the most enriched pathway in all the three pathologic variants of cervical cancer was the "ubiquitin mediated proteolysis" pathway. Eight key candidate circRNAs derived from this pathway were further validated, and we noticed that several target miRNAs of candidate circRNAs could target the source genes. Based on this we constructed a related competing endogenous RNA (ceRNA) network. Conclusion: Through a comprehensive interpretation of differentially expressed circRNAs in different pathologic variants of cervical cancer, this study provides new insights into the process of tumor differentiation mediated by HPV. Our results may help to complement the molecular typing and stem cell theory of cervical cancer.

miR-125a attenuates the malignant biological behaviors of cervical squamous cell carcinoma cells through Rad51.

Cervical squamous cell carcinoma (CSCC), the most common cervical malignancy, is more likely to invade and metastasize than other cervical cancers. miR-125a, a tumor suppressor gene, has been confirmed to be associated with cancer metastasis. However, the role of miR-125a in CSCC and the underlying mechanism are unknown. miR-125a expression was confirmed by real-time quantitative PCR (RT-qPCR), and the Rad51 expression level was measured by western blotting analysis. CSCC cell proliferation, migration and invasion were assessed with functional assays, including CCK-8, colony formation, wound healing and Transwell assays. Our data confirmed that miR-125a is expressed at low levels in CSCC tissues and cells. Functionally, the overexpression of miR-125a greatly prevented the proliferation, migration and invasion of CSCC cells, and the inhibition of miR-125a expression strongly enhanced these behaviors in CSCC cells. Moreover, the expression of Rad51, a miR-125a target gene, greatly reversed the miR-125-mediated inhibition of CSCC cell proliferation, migration and invasion. In addition, we discovered that miR-125a downregulated the levels of phosphorylated PI3K, AKT and mTOR through Rad51 in CSCC cells. miR-125a, a tumor suppressor, can attenuate the malignant behaviors of CSCC cells by targeting Rad51. Therefore, the miR-125a/Rad51 axis might be a target for CSCC therapy.

Effects of Forkhead box O1 on lipopolysaccharide-induced mitochondrial dysfunction in human cervical squamous carcinoma SiHa cells.

Persistent infection and chronic inflammation play important roles in the development of cervical squamous cell carcinoma. Forkhead box O1 (FOXO1) is a notable regulator of mitochondrial metabolism, which is involved in the occurrence and development of tumors. The present study explored the effects of FOXO1 in human cervical squamous carcinoma SiHa cells. The expression of FOXO1 was examined using reverse transcription-quantitative PCR, western blotting and immunohistochemical staining. SiHa cell migration and proliferation were detected using Transwell and 3H-TdR assays. Mitochondrial functions were assessed based on reactive oxygen species (ROS) generation and changes in the mitochondrial membrane potential (ΔΨm). The present study revealed that lipopolysaccharide (LPS) stimulation significantly inhibited the expression of FOXO1 in cervical squamous carcinoma SiHa cells; while silencing FOXO1 resulted in the accumulation of mitochondrial ROS, a decrease in the ΔΨm and abnormal morphology of mitochondria. Accordingly, enhancing FOXO1 expression or treatment with metformin, which protects mitochondrial function, reversed LPS-induced mitochondrial dysfunction, cell pyroptosis, migration and proliferation of cervical squamous carcinoma SiHa cells. Overall, the current study indicated that treatment with FOXO1 could potentially be used as therapeutic strategy to prevent LPS-induced cervical squamous cell carcinoma-related dysfunction in a mitochondria-dependent manner.

Reduction in immune cell number and loss of 5hmC are associated with lesion grade in cervical carcinogenesis.

Tumor genome methylation is closely related to tumor immunosuppression. In the present study, we evaluated the fluctuations in DNA methylation levels, and the numbers of infiltrating T cells and their cytokines in different-grade cervical lesions. A total of 154 human cervical specimens that included LSIL (43 cases), HSIL (48 cases), and cervical squamous cancer (63 cases) were used for this study. Immunohistochemistry for 5-hydroxymethylcytosine (5hmC) and T-cell-attracting chemokines was performed, and multiplex immunofluorescence labeling was used to identify different T-cell subtypes. We found that the proportions of samples that immunostained weakly or negatively for 5hmC were increased commensurately with elevations in the severity of cervical lesions. The expression of T-cell-attracting chemokines-including CXCL9, CXCL10, and CXCL11-was positively associated with 5hmC levels, and CXCL9 was the cytokine that was most pronounced. With the progression of cervical lesions, the numbers of total T cells, CTL, and NK cells in the cervical tissues all gradually decreased. During the occurrence and development of cervical squamous carcinoma, 5hmC was gradually lost, and immunosuppression occurred in precancerous cervical lesions.

Study on Reversal Effect of Quercetin on Human Cervical Squamous Carcinoma Cisplatin-resistant Cell Line SiHa/DDP / 中国药房

OBJECTIVE：To study the reversal effect of quercetin on human cervical squamous carcinoma cisplatin-resistant cell line SiHa/DDP. METHODS ：The drug resistance index of cisplatin to SiHa/DDP cells ，and the reversal resistance multiple of quercetin to SiHa/DDP cells were determined. The effects of quercetin （0.005 μg/mL），cisplatin（2.5 μg/mL），cisplatin combined with quercetin （2.5 μg/mL cisplatin+0.005 μg/mL quercetin），quercetin combined with pathway inhibitor（0.005 μg/mL quercetin+ 20 nmol/L rapamycin ）on the apoptotic rate of SiHa/DDP cells were investigated ，as well as its effects on the expression of phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian rapamycin target protein （mTOR） signaling pathway related proteins （PI3K，Akt，mTOR，P-gp，p70S6K）. RESULTS ：The resistance index of cisplatin to SiHa/DDP cells was 5.19， and reversal resistance multiple of quercetin to SiHa/DDP cells was 4.00. Compared with cisplatin alone and quercetin alone ， cisplatin combined with quercetin ，quercetin combined with rapamycin could significantly increase the apoptotic rate of SiHa/DDP cells（P＜0.05），while decreased the phosphorylation of Akt ，mTOR and p 70S6K protein as well as the expression of P-gp protein （P＜0.05）. CONCLUSIONS ：Quercetin can effectively reverse drug resistance of SiHa/DDP cells to cisplatin ，which may be associated with inhibiting the expression of the protein related to PI 3K/Akt/mTOR signaling pathway.

Identification of significant genes signatures and prognostic biomarkers in cervical squamous carcinoma via bioinformatic data.

BACKGROUND: Cervical squamous cancer (CESC) is an intractable gynecological malignancy because of its high mortality rate and difficulty in early diagnosis. Several biomarkers have been found to predict the prognose of CESC using bioinformatics methods, but they still lack clinical effectiveness. Most of the existing bioinformatic studies only focus on the changes of oncogenes but neglect the differences on the protein level and molecular biology validation are rarely conducted. METHODS: Gene set data from the NCBI-GEO database were used in this study to compare the differences of gene and protein levels between normal and cancer tissues through significant pathway selection and core gene signature analysis to screen potential clinical biomarkers of CESC. Subsequently, the molecular and protein levels of clinical samples were verified by quantitative transcription PCR, western blot and immunohistochemistry. RESULTS: Three differentially expressed genes (RFC4, MCM2, TOP2A) were found to have a significant survival (P < 0.05) and highly expressed in CESC tissues. Molecular biological verification using quantitative reverse transcribed PCR, western blotting and immunohistochemistry assays exhibited significant differences in the expression of RFC4 between CESC and para-cancerous tissues (P < 0.05). CONCLUSION: This study identified three potential biomarkers (RFC4, MCM2, TOP2A) of CESC which may be useful to clarify the underlying mechanisms of CESC and predict the prognosis of CESC patients.

An EGFR-Amplified Cervical Squamous Cell Carcinoma Patient with Pulmonary Metastasis Benefits from Afatinib: A Case Report.

Currently, women with metastatic or recurrent cervical cancer still have very limited treatment options. Despite the rapid advancements in targeted therapies, no targeted therapy was approved for cervical cancer, except for bevacizumab. In the present study, we reported a 52-year-old heavily pre-treated EGFR amplified patient with metastatic cervical squamous cancer who benefited from afatinib with a progression-free survival (PFS) of 5.5 months. The patient was administered with a first-line treatment of chemotherapy and bevacizumab with a PFS of 4.3 months. Subsequently the patient was treated with a second-line regimen of angiogenesis inhibitor apatinib plus chemotherapy and a third-line treatment of pembrolizumab. Genomic profiling revealed significant EGFR amplification in both primary (copy number [CN] =15.9) and metastatic lesions (CN =18). Afatinib monotherapy was then administered as the fourth-line regimen. She achieved partial response (PR) with a PFS of 5.5 months. At disease progression, the CN of EGFR was elevated to 39.9 accompanied by the emergence of PIK3CA amplification (CN =4.2). The patient was treated with everolimus and afatinib and achieved stable disease (SD) after 3 months. To the best of our knowledge, this is the first clinical evidence of an EGFR-amplified metastatic cervical cancer patient benefiting from afatinib as a single agent.

Using amide proton transfer to identify cervical squamous carcinoma/adenocarcinoma and evaluate its differentiation grade.

PURPOSE: To explore the possibility of using amide proton transfer-weighted imaging (APTWI) for the identification and diagnosis of cervical squamous carcinoma (CSC), cervical adenocarcinoma (CA) and different levels of CSC. MATERIALS AND METHODS: Seventy-six patients with newly diagnosed uterine cervical cancer (UCC) were studied prior to treatment, including 20 with poorly differentiated (Grade 3) CSC, 23 with moderately differentiated (Grade 2) CSC, 17 with well-differentiated (Grade 1) CSC, and 16 with CA (13 with poorly differentiated (Grade 3) CA and 3 with moderately differentiated (Grade 2) CA). Differences in the magnetization transfer ratio at 3.5 ppm (MTRasym (3.5 ppm)) were identified between CSC and CA and between high-level (Grade 3) CSC and low-level (Grade 2 and Grade 1) CSC, as well as among all three grades of CSC differentiation. Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic thresholds and performance of the parameters. Spearman correlation analysis was used to examine the correlation between the MTRasym (3.5 ppm) and histological grade. RESULTS: The MTRasym (3.5 ppm) in CA was higher than that in CSC (P = 0.001). The MTRasym (3.5 ppm) in high-level CSC was higher than that in low-level CSC (P = 0.001). The MTRasym (3.5 ppm) was positively correlated with the grade of CSC differentiation (r = 0.498, P = 0.001). The MTRasym (3.5 ppm) in Grade 3 CSC was higher than that in Grade 2 and Grade 1 CSC (P = 0.02/0.01). No significant difference in the MTRasym (3.5 ppm) was found between Grade 2 CSC and Grade 1 CSC (P = 0.173). The area under the ROC curve (AUC) for the MTRasym (3.5 ppm) in distinguishing CSC and CA was 0.779, with a cut-off, sensitivity, and specificity of 2.97%, 60.0% and 82.5%, respectively. The AUC for distinguishing high-/low-level CSC was 0.756, with a cut-off, sensitivity, and specificity of 3.29%, 68.8% and 83.3%, respectively. CONCLUSION: APTWI may be a useful technique for the identification and diagnosis of CSC, CA and different levels of CSC, which may have an important impact on clinical strategies for treating patients with UCC.

Imaging Features of the Whole Uterus Volume CT Perfusion and Influence Factors of Blood Supply: A Primary Study in Patients with Cervical Squamous Carcinoma.

RATIONALE AND OBJECTIVES: To explore the imaging features of whole uterus volume CT perfusion (vCTP) and the influence factors of blood supply in cervical squamous carcinoma (CSC). MATERIALS AND METHODS: vCTP was performed on a 640-slice computed tomography system in 43 patients with CSC diagnosed by biopsy, and 24 cases of them underwent magnetic resonance imaging. The size of the tumor was measured on vCTP and magnetic resonance (MR) images. Perfusion parameters, including arterial blood flow (AF), blood volume, and permeability surface (PS), were measured by two radiologists, using interclass correlation coefficient to evaluate the interobserver reliability. The difference of tumor size and perfusion data was analyzed by paired t test and rank sum test. The correlation of perfusion parameters with some factors was analyzed by Pearson or Spearman correlation analysis. RESULTS: Tumor sizes were not significantly different between vCTP and MR images. The interclass correlation coefficient of each parameter was 0.818-0.945. The AF value of CSC was significantly higher than normal uterine body, and the blood volume and PS values of CSC were not statistically different compared with those of normal uterine body. There was no significant difference in AF value of CSC among different FIGO stages and pathological grades. The AF and PS values of CSC were negatively correlated with the age of the patients. CONCLUSION: The vCTP could accurately shows the size of the CSC with use of MR as the reference standard, and its perfusion parameters have good measurement stability; the CSC was hypervascular, but this trend was less pronounced in older women.

The role of eukaryotic translation initiation factor 5A-1 (eIF5A-1) gene in HPV 16 E6 induces cell growth in human cervical squamous carcinoma cells.

Human Papillomavirus (HPV) is considered as the major risk factor for the development and progression of cervical cancer. The high expression of HPV 16 E6 may be the causative factor for induction and maintenance of the transformed phenotype. These oncoproteins would interact with several intracellular proteins, such as eukaryotic translation initiation factor 5A-1 (eIF5A-1) that is essential for proliferation of eukaryotic cells. Our study explored the expression level of HPV 16 E6 and eIF5A-1 in human cervical squamous carcinoma samples and the adjacent non-cancerous cervix samples. Both C33a cells and SiHa cells transfected with a vector encoding HPV 16 E6 resulted in increase of eIF5A-1 expression level and enhancement of viability, migration and proliferation of these cells, furthermore, these effects in both C33a cells and SiHa cells could be abrogated by treatment with eIF5A-1 small-interfering RNA (siRNA) or the specific inhibitors ciclopirox (CPX) that was used to inhibit the function of eIF5A-1 via blocking the main enzymes deoxyhypusine hydroxylase (DOHH). Our results support that the silencing the eIF5A-1 gene or blocking the DOHH could induce the apoptosis of HPV 16 E6-infected cervical carcinoma cells. Thus might provide a new approach to preventing and treating cervical cancer.

MiR-22-3p Regulates Cell Proliferation and Inhibits Cell Apoptosis through Targeting the eIF4EBP3 Gene in Human Cervical Squamous Carcinoma Cells.

Background: MicroRNAs (miRNAs) are non-coding small RNAs that function as negative regulators of gene expression and are involved in tumour biology. The eIF4E-binding proteins (eIF4EBPs) play essential roles in preventing translation initiation and inhibiting protein synthesis at a global or message-specific level in a variety of tumours. Methods: According to comparative miRNA profiles of clinical cervical cancer and non-cancerous cervical tissue specimens, several miRNAs were aberrantly expressed in the cervical cancer samples. C33a and SiHa cell proliferation and apoptosis were detected using methyl thiazolyl tetrazolium (MTT) and flow cytometry assays, respectively. Results: Among the aberrantly expressed miRNAs, miR-22-3p was significantly differentially expressed in cervical cancer tissues and was highly associated with cervical cancer cell growth regulation. In addition, bioinformatic predictions and experimental validation were used to identify whether eIF4E-binding protein 3 (eIF4EBP3) was a direct target of miR-22-3p; eIF4EBP3 protein levels were generally low in the cervical cancer tissues. Furthermore, functional studies revealed that either a miR-22-3p inhibitor or eIF4EBP3 overexpression could induce apoptosis in cervical cancer cells in vitro. Importantly, we found that eIF4EBP3 accumulation could significantly attenuate cervical cancer cell proliferation triggered by a miR-22-3p mimic as well as enhance apoptosis in cervical cancer cells. Conclusion: Taken together, our data provide primary proof that miR-22-3p can induce cervical cancer cell growth at least in part by up-regulating its expression to decrease eIF4EBP3 expression levels; miR-22-3p thus holds promise as a prognostic biomarker and potential therapeutic target for treating cervical cancer.

Rapid diagnosis of cisplatin-sensitive and resistant cervical squamous cell carcinomas by reverse transcription loop-mediated isothermal amplification.

Cisplatin is a widely used platinum anticancer drug in cervical cancer chemotherapy; however, its use is limited by the development of drug resistance, which is often rationalized via effects on cellular uptake. Human copper transporter 1 (hCTR1) was proven as crucial regulator for cellular platinum uptake in cancer cells and improving effect of cisplatin treatment. Thus, methods to detect expression of hCTR1 in patients in biopsy specimens with certain rapidity and accuracy are essential to evaluate platinum drug sensitivity and perform further individualized treatment. In this study, a total of 46 normal cervical and primary cervical squamous cell carcinoma tissues were collected and tested by reverse transcription loop-mediated isothermal amplification (RT-LAMP) analysis targeted hCTR1 mRNA. Sensitivity and specificity of RT-LAMP results were calculated and compared to immunohistochemical (IHC) examination. Overexpression of hCTR1 RNA was detected in 12 samples using RT-LAMP within 45 minutes reaction, which indicated positivity with cisplatin resistance. This new technique is suggested to be alternative for rapid diagnosis of cisplatin sensitive patients whose hCTR1 is prevalent.

Pembrolizumab in recurrent advanced cervical squamous carcinoma.

No definitive cure is known for recurrent, persistent or metastatic cervical carcinoma. Chemotherapy remains the standard of care, although available options are scarce and do not provide satisfactory results. Immune checkpoint inhibitors have shown a strong and prolonged response in several types of cancer, although only in a subset of patients. Defining the profile of the patients likely to benefit from such treatment is a subject of active research. Here, we present a case of a heavily pretreated patient with recurrent advanced squamous cell carcinoma of the cervix who had exhausted all available treatment options and showed a striking response to the immune checkpoint inhibitor pembrolizumab.

Expressions of Clusterin and E-cadherin in cervical squamous carcinoma and their correlation / 西安交通大学学报(医学版)

Objective To explore the expressions of clusterin and E-cadherin in cervical squamous tissues and their correlation.Methods Immunohistochemical technique was used to measure the expressions of clusterin and E-cadherin in 18 normal cervical tissues,32 cervical intraepithelial neoplasia (CIN)tissues and 40 cervical squamous carcinoma (CSC)tissues.Results There were on significant differences in baseline characteristics among the three groups based on standardized differences.The expressions of clusterin in CSC,CIN2 and CIN3 tissues were significantly higher than those in normal cervical tissues (P <0.05 ).The expression of clusterin was significantly lower in grade Ⅰ than in grade Ⅲ (P <0.01)and was connected with clinical stage (P <0.05 )instead of lymph node metastasis and cancer stromal invasion depth (P > 0.05 ).The expressions of E-cadherin in tissues of CSC, CIN2 and CIN3 were significantly lower than those in normal cervical tissues (P <0.05).E-cadherin expression was both significantly higher in grade Ⅰ and grade Ⅱ than in grade Ⅲ (P <0.05),and was associated with lymph node metastasis (P < 0.05 )rather than clinical stage and cancer stromal invasion depth (P > 0.05 ).A negative correlation was found between the expressions of clusterin and E-cadherin in CSC (r = - 0.339,P < 0.05 ). Conclusion The significantly different expressions of clusterin and E-cadherin may be related to the development of CSC,suggesting that the two may be used as potential markers for early diagnosis of CSC.

Expressions of Clusterin and E-cadherin in cervical squamous carcinoma and their correlation / 西安交通大学学报(医学版)

Objective To explore the expressions of clusterin and E-cadherin in cervical squamous tissues and their correlation.Methods Immunohistochemical technique was used to measure the expressions of clusterin and E-cadherin in 18 normal cervical tissues,32 cervical intraepithelial neoplasia (CIN)tissues and 40 cervical squamous carcinoma (CSC)tissues.Results There were on significant differences in baseline characteristics among the three groups based on standardized differences.The expressions of clusterin in CSC,CIN2 and CIN3 tissues were significantly higher than those in normal cervical tissues (P <0.05 ).The expression of clusterin was significantly lower in grade Ⅰ than in grade Ⅲ (P <0.01)and was connected with clinical stage (P <0.05 )instead of lymph node metastasis and cancer stromal invasion depth (P > 0.05 ).The expressions of E-cadherin in tissues of CSC, CIN2 and CIN3 were significantly lower than those in normal cervical tissues (P <0.05).E-cadherin expression was both significantly higher in grade Ⅰ and grade Ⅱ than in grade Ⅲ (P <0.05),and was associated with lymph node metastasis (P < 0.05 )rather than clinical stage and cancer stromal invasion depth (P > 0.05 ).A negative correlation was found between the expressions of clusterin and E-cadherin in CSC (r = - 0.339,P < 0.05 ). Conclusion The significantly different expressions of clusterin and E-cadherin may be related to the development of CSC,suggesting that the two may be used as potential markers for early diagnosis of CSC.

Viral DNA load of high-risk human papilloma virus is closely associated with the grade of cervical lesions.

This study is to explore the correlation between the viral load of high-risk human papilloma virus (HPV) and the degree of cervical lesions, as well as the follow-up monitoring role of high-risk HPV measurements in the treatment of patients with cervical lesions. Hybrid capture-2 method was used to measure the amount of high-risk HPV load of 361 patients who were enrolled from January 2009 to December 2010 at the Affiliated Tumor Hospital of Xinjiang Medical University, including 76 cases of cervical squamous carcinoma, 119 cases of cervical intraepithelial neoplasia and 166 cases of cervicitis. The correlation between the viral load of high-risk HPV and the degree of cervical lesions was analyzed using correlation analysis. Patients with cervical intraepithelial neoplasia (CIN) and cervical squamous carcinoma were followed up until December 2013, with the follow-up time being 37-60 months. Statistically significant differences in the high-risk HPV load existed between cervicitis group, CIN group and cervical squamous carcinoma group (P = 0.000). In addition, the viral load was increased with the increase of the severity of cervical lesions, showing a positive correlation (r = 0.436, P = 0.000). During the follow-up, 6 cases of vaginal intraepithelial neoplasia, 3 cases of recurrence CIN and 1 case of vaginal squamous cell carcinoma of the vulva were found, which were shown to relate with the continuing high-risk HPV infection in vagina. Viral load of high-risk HPV were positively correlated with the severity of cervical lesions, playing an important role in the monitoring of patients with cervical lesions after treatment.

Expression and clinical significance of γH2 AX in HPV16 positive cervial squamous carcinoma / 中华微生物学和免疫学杂志

Objective To investigate the expression and significance of γH2AX in cervical squamous carcinoma.Methods Firstly,DNA were extracted from 74 cervical squamous carcinoma samples and PCR were tested for HPV infection.Secondly,formalin-fixed paraffin-embedded tissue sections (4 μm)were stained with H&E method to detect cervical lesions grading.Thirdly,HPV16 DNA were examined by in situ hybridization(ISH) and γH2AX,p16 were examined by immunohistochemical (IHC) staining.Then,30 cases typical tissue sections in which including the normal cervical tissue,cervical intraepithelial neoplasia and cervical carcinoma in situ were selected for comparing the HPV DNA loading,and the γH2AX and,pl6 expression.Finally,the feasibility of γH2AX serving as a biomarks in HPV infection-related cervical carcinogenesis were analyzed.Results In this study,HPV infection ratio is 98.65％,and HPV16 is the most common type with 74.32％ infection.In situ hybridization showed no HPV16 DNA exist in normal cervical tissues and CINI.In CIN Ⅱ HPV DNA exist mainly as episomal DNA.With the increasing of cervical lesions grade,HPV DNA was integrated into chromosome steadily.The expression of γH2AX and pl6 were positively associated with grading of cervical lesions.HPV DNA and γH2AX protein co-exist primarily in the prickle cell layer and the granular cell layer.The HPV DNA and p16 protein exist in different cell layer.Conclusion γH2AX may be employed as a biomarker for HPV positive cervical carcinogenesis.

Implications of continued upregulation of p16INK4a evolution from high-grade squamous intraepithelial lesion to invasive squamous carcinoma of the cervix

On integration into the host cervical keratinocyte genome, human papillomavirus (HPV) E7 protein binds pRB, releasing E2F from normally incompetent pRB-E2F complexes and allowing propagation of G1-S transition by the E2F. p16 INK4a, a tumour suppressor protein, increases in refl ex response to counter this. 29 histologically re-confi rmed low-grade squamous intraepithelial lesions (LSIL), 27 high-grade squamous intraepithelial lesions (HSIL) and 30 invasive cervical squamous carcinoma (SCC) were immunohistochemically stained for p16INK4a expression using the CINtec Histology Kit (REF 9511, mtm laboratories AG, Heidelberg, Germany) to re-affi rm the notion that integration of HPV occurs predominantly in SCC and possibly HSIL and less in LSIL and normal squamous epithelium (NSqE). Implicit was also the attempt to understand the role of E2F, as indicated by p16INK4a, in evolution of SCC from HSIL. No ethnic predilection was noted for LSIL, HSIL or SCC. Patients with SCC were signifi cantly older by about 14-years compared with HSIL (p<0.05) while there was no signifi cant age difference between HSIL and LSIL. p16INK4a expression was signifi cantly increased (p<0.05) in both HSIL (88.9%) and SCC (83.3%) compared with LSIL (3.4%) and NSqE (0%); the NSqE being normal squamous epithelium noted in 17 of the LSIL, 19 HSIL and 5 SCC. From these fi ndings there is suggestion that fundamental upstream events viz HPV integration, E7 upregulation followed by E2F activation occurs at point of transformation to HSIL and continues unrelentingly for another one to two decades before hitherto unclear factors convert a non-invasive lesion into an overtly invasive malignant counterpart. Interestingly, the occurrence of HSIL and LSIL in almost the same age group could mean that alteration from episomal to integrated form of HPV may not incur a prolonged incubation period, unlike from HSIL to SCC