Normal Cervix Research Articles

OP10.03: Gestational age‐related changes in uterine cervical elasticity in normal pregnancy at 12–35 weeks' gestation evaluated using shear wave elastography

To analyse the gestational age-related changes in elasticity of the normal uterine cervix at 12-35 weeks' gestation using shear wave elastography. The subjects consisted of 417 cases of normal singleton pregnancy at 12-35 weeks' gestation, cared for in our hospital from July 2016 to January 2017. The shear wave speed (SWS) of the cervix was measured using transvaginal ultrasonography (Aplio 500, Toshiba). On the sagittal section, sampling points were placed on the anterior (A), posterior (P), and cervical canal (C) regions of the internal and external os (IO/EO). After dividing the observational period into 6 groups at 4-week interval, the following parameters were analysed: 1) the changes in SWS at each point with advancing gestation; 2) comparison of SWSs at both the IO and EO for A, P and C. Statistical analyses were made using piecewise linear regression analysis, the Mann-Whitney U-test, the Kruskal-Wallis and the Wilcoxon test as post-hoc test. Significance was set at P<0.05. The SWSs of posterior at the IO decreased in piecewise linear regression, with a critical point at 20 weeks, whereas the SWSs of the remaining points decreased linearly. At the IO, the SWSs of P were higher than those of A and C at all age-groups. The SWSs of P and A at the IO were higher than those at the EO at all age-groups. The SWSs of C at the IO were higher than those at the EO before 20-23 weeks, respectively, however, the SWSs were not different between the points thereafter. The SWSs of A at the IO were higher than those at the EO before 28-31 weeks, respectively, however, the SWSs were not different between the points thereafter. The posterior cervix at the internal os, the hardest point of the cervix at all age-groups, softened with advancing gestation, while the endocervix was harder than the ectocervix before 35 weeks, showing a physiological cervical maturation with advancing gestation.

Interaction between polycyclic aromatic hydrocarbons and p16, FHIT gene CpG island methylation in patients with cervical intraepithelial neoplasias

Objective: To explore the effect of polycyclic aromatic hydrocarbons (PAHs) and p16, FHIT gene CpG island methylation, as well as their interaction in cervical intraepithelial neoplasias. Methods: Objects of this study were from a cohort of cervical lesions study in Yangqu county of Shanxi province. All the patients were diagnosed pathologically, that including 83 patients with high-grade cervical intraepithelial neoplasia (CINⅡ/Ⅲ), 86 patients with low-grade cervical intraepithelial neoplasia (CINⅠ) and another 91 women under normal cervical (NC) condition. 1-hydroxy pyrene in the urine was detected by high performance liquid chromatography (HPLC) while CpG island methylation status of tumor suppressor gene p16 and FHIT were measured by methylation-specifc polymerase chain reaction (MSP). Data were analyzed with Kruskal-Wallis H test, chi-square test and trend of chi-square test. Logistic regression models were used to estimate the odds ratio (OR) and corresponding 95% confidence intervals (95%CI) between influencing factors and the cervical disease by using the SPSS statistical software (version 20.0). The interaction under study was evaluated by using the generalized multifactor dimensionality reduction (GMDR) model. Results: Level of 1-hydroxy pyrene (H=50.743, P<0.001) and the high exposure rate of 1-hydroxy pyrene (trend χ(2)=20.146, P<0.001) were gradually increasing along with the severity of cervical intraepithelial neoplasia. The CpG island methylation rates of p16, FHIT in CINⅠand CINⅡ/Ⅲ group were higher than that in NC group, and gradually increasing along with the severity of cervical intraepithelial neoplasia (trend χ(2)=9.75, P=0.002; trend χ(2)=10.39, P=0.001). Results from the GMDR model showed that interaction existed among the high exposure of 1-hydroxy pyrene and the CpG island methylation of p16, FHIT in CINⅠ and CINⅡ/Ⅲ group. Conclusion: Under the high exposure of 1-hydroxy pyrene and the CpG island methylation of p16, FHIT appeared to have increased the risk of cervical intraepithelial neoplasia and causing synergistic effect in cervical intraepithelial neoplasia.

Retinol as a micronutrients related to cervical local immunity: The expression of tumor necrosis factor-alpha specifically stimulated with E6 epitope of human papillomavirus type-16 and ratio of CD4+/CD8+ T cell in natural history of cervical cancer

Retinol is one of the antioxidant micronutrients that plays essential roles in the immune system, by preventing the persistence of modulating CD4+ and CD8+ T cells and cytokines production. Tumor Necrosis Factor-Alpha (TNF-α) is an acute pro-inflammatory cytokine which has many crucial roles in controlling HPV. In contrast, when persistent infection occurs, TNF-α induces carcinogenesis. The ratio of CD4+ cells to CD8+ T cells and adequate TNF-α production in acute HPV infection are key points for clearance. The aim of this research is to analyze the sufficiency level of retinol deposit, the expression of TNF-α, and the ratio of CD4+: CD8+ T cells in a normal cervix, clearance and persistent HPV subclinical infection, and cervical cancer group. The sufficiency level of retinol deposit was analyzed from peripheral blood using the ELISA method. The cervico-vaginal secretions, which were incubated for 24 hours, were stimulated specifically by E6 epitope HPV type-16, measuring TNF-α expression semi-quantitatively by the ELISpot method and CD4+/CD8+ T cells quantitatively by flowcytometry method. The sufficient level of retinol deposit in a normal cervix, clearance HPV subclinical infection, persistent, and cervical cancer group was 85%, 75% (OR 1.89), 33.3% (OR 11.33), and 75% (OR 1.89), respectively. The expression of TNF-α in normal cervix group was 10%, while for cervical cancer it was 75% (OR 27.00; p < 0.001). There was no expression in clearance and persistent HPV subclinical infection groups. A high ratio of CD4+: CD8+ T cells in the normal cervix and cervical cancer group was 10% and 25% (OR 0.33). There was no high ratio of CD4+: CD8+ T cells in clearance (OR 1.22) and persistent (OR 0.95) HPV subclinical infection groups. This study was able to prove that the normal cervix group has the highest retinol deposit sufficiency level and the cervical cancer group has the highest TNF-α expression (OR 27; p < 0.001). The lowest of retinol deposit sufficiency level was not in cervical cancer, but in the persistent HPV subclinical infection group (OR 11.33). There was significant correlation in TNF-α expression between cervical cancer and normal cervix (p < 0.001), cervical cancer and clearance subclinical HPV infection (p = 0.024), and between clearance and persistent group (p = 0.007).

Fertility and the Reproductive Tract in a Woman with Caudal Duplication Syndrome

Abstract Background: Caudal duplication syndrome is a rare entity characterized by complete duplication of the genitourinary system and hindgut structures. There have been 13 case reports of women with genitourinary duplication; only 3 had complete caudal duplication and only 1 had a successful term pregnancy. There are no case reports to date of patients having more than 1 successful pregnancy with this syndrome. Case: A 28-year-old multiparous woman with known caudal duplication syndrome presented with recurrent urinary tract infections and hydronephrosis. This patient was interested in having a unification of her two vaginas. An examination under anesthesia revealed two gluteal clefts, three labia minora, two vaginas widely separated by >4 cm, a bifid clitoris, two normal cervices, two urethras, a right perforate anus, and a left imperforate anus. Exploratory laparotomy revealed uterine didelphys, 2 bladders, 2 ureters, and 2 kidneys. Results: Excision of the left bladder and ureteral reimplantation re...

PREVALENCE OF HUMAN PAPILLOMA VIRUS TYPES 16 AND 18 AMONG WOMEN WITH CERVICAL CANCER IN ALEPPO, SYRIA

Objective: Cervical cancer is the second common type of cancer among women worldwide. Infection by certain types of human papillomavirus infection (HPV) such as HPV 16/18 has been considered as the most significant risk factor for the development of cervical cancer. This study was conducted to determine the prevalence HPV 16/18 in women with and without cervical cancer in Aleppo city, Syria.Methods: Paraffin embedded cervical tissue samples were collected from 113 patients and 40 controls (normal cervix), DNA was extracted and analyzed by PCR for the presence of HPV16/18. Out of 113 cases of cervical carcinoma, 25 cases (22.1%) were carcinoma in situ (CIS), 72 cases (63.7%) were squamous cell carcinomas (SCC) and 16 cases (14.2%) were adenocarcinoma (ADC).Results: HPV 16 and/or 18 DNA was detected in 84 (74.3%) cases out of 113 of cervical cancers, while it was found in only two of the 40 samples in the control group. At the same time, 9 out of 25 (36%) carcinoma in situ, 61 out of 72(84.72%) squamous cell carcinomas, and 14 out of 16 (87.5%) adenocarcinomas, were positive for HPV 16 and/or 18.Conclusion: The frequency of infection with HPV 16 and 18 is high among Syrian women and a vaccine against these two types may be effective in about 74% of cervical cancers in Syria.

Intravoxel Incoherent Motion Diffusion-Weighted Magnetic Resonance Imaging of Cervical Cancer With Different b-Values.

The aims of this study were to evaluate the dependence of diffusion parameters on the b values adopted for intravoxel incoherent motion diffusion-weighted magnetic resonance imaging and to investigate the application value of multiple diffusion parameters obtained from monoexponential and biexponential models in subjects with a normal cervix and in cervical cancer patients. A total of 120 female patients with cervical cancer and 21 female control subjects with a normal cervix underwent diffusion-weighted magnetic resonance imaging with 13 b values (0-2000 s/mm) at 3 T. The standard apparent diffusion coefficient (Dst), diffusion coefficient (D), pseudodiffusion coefficient (D*), and perfusion fraction (f) were calculated by fitting with monoexponential and biexponential models at 2 different ranges of b values: 0 to 1000 and 0 to 2000 s/mm. A univariate analysis was performed to identify factors that could distinguish cervical carcinoma from normal cervical tissue. Parameters that correlated with the pathological grade and stage of cervical cancer were also evaluated. Receiver operating characteristic curves were used to evaluate the diagnostic efficiency of every parameter. All the tested parameters, except the D* of the 2 different ranges of b value groups, significantly differed between the patients with cervical carcinoma and control subjects (P < 0.01). D2000, Dst2000, and D1000 showed comparable diagnostic value, with an area under the curve of 0.923, 0.909, and 0.907, respectively. Dst2000, D2000, Dst1000, and D1000 differed significantly among the 3 degrees of cervical stromal infiltration depth (P < 0.05). D2000 and Dst2000 tended to outperform D1000 in terms of diagnostic efficiency, but there was no significant difference in their ability to differentiate cervical carcinoma from normal cervix. Cervical cancers with lower Dst and D values tended to have greater infiltration depth.

Abstract 1866: Ultrahigh resolution optical coherence microscopy for label-free imaging of human cervical lesions ex vivo

Abstract Cervical cancer remains the fourth most common cause of cancer and the fourth leading cause of cancer death in women worldwide. In contrast to the decreasing prevalence in developed countries, incidence of cervical cancer remains high in developing countries, such as China, India and Chile. Effective screening of cervical cancer is crucial for early diagnosis of the disease. Ultrahigh resolution optical coherence microscopy (UHR OCM) is an emerging label free imaging technique that can be used to obtain three dimensional (3D) “optical biopsy” of biological samples with cellular resolution up to 2 mm beneath the tissue surface. The goal of this study is to assess the diagnostic value of UHR OCM for human cervical precancer and cancer screening and identify characteristic features in UHR OCM images that correlate with the gold standard histopathology. Six hundred and fifty eight (658) fresh cervical tissues from cervical biopsy, conization and panhysterectomy were obtained from 254 consenting volunteers. Multiple 3D UHR OCM images were obtained from each specimen before the sample was processed for standard histopathology evaluation. Of the 658 samples imaged, 474 had normal cervix or benign lesions, 110 had low grade squamous intraepithelial lesion (LSIL), 57 had high grade squamous intraepithelial lesions (HSIL) and 17 had invasive cancer. UHR OCM images were compared to histopathology slides obtained from the same specimens in order to identify diagnostic image features. Distinctive patterns for squamous epithelium, columnar epithelium, basement membrane, cervical inflammation and erosion, koilocytic cells, low grade and high grade squamous intraepithelial lesions and cervical cancer were clearly observed. Our results lay the foundation for future non-invasive optical evaluation of cervical tissue in vivo. This could lead to a less invasive and more effective screening strategy for cervical cancer based on OCM guided biopsies. Citation Format: Xianxu Zeng, Canyu Li, Xiaofang Wang, Jason Jerwick, Tao Xu, Yuan Ning, Linlin Zhang, Yutao Ma, Xiaoan Zhang, Chao Zhou. Ultrahigh resolution optical coherence microscopy for label-free imaging of human cervical lesions ex vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1866. doi:10.1158/1538-7445.AM2017-1866

Level of Retinol Deposit and Cervical Cancer

Objective: To analyze level of retinol deposit sufficiency in the&#x0D; natural history of cervical cancer.&#x0D; &#x0D; Methods: Serum retinol level was measured by ELISA from&#x0D; peripheral blood of subjects with normal cervix, cleared and&#x0D; persistent high risk human papilloma virus (HR-HPV) subclinical&#x0D; infection, and cervical cancer who fulfilled the inclusion and&#x0D; exclusion criteria. The study was held in Dr. Cipto Mangunkusumo&#x0D; and Fatmawati Hospital, Jakarta, within 2 years (August 2013-&#x0D; 2015). Blood was taken twice, consisting of post-8-hour fasting&#x0D; blood and 2 hours after 6000 IU retinyl palmitate oral&#x0D; administration.&#x0D; &#x0D; Results: Of 47 total samples, sufficient level of retinol deposit in&#x0D; normal cervix, cleared and persistent HR-HPV subclinical infection,&#x0D; and cervical cancer group was 85.0% (reference), 75.0% (OR 1.89),&#x0D; 33.3% (OR 11.33), and 75% (OR 1.89); respectively. Statistically,&#x0D; there was no significant difference from sufficiency level of retinol&#x0D; deposit between normal cervix and clearance HR-HPV subclinical&#x0D; infection (p=0.628), normal cervix and persistent HR-HPV&#x0D; subclinical infection (p=0.078), normal cervix and cervical cancer&#x0D; (p=0.433), cervical cancer and clearance HR-HPV subclinical&#x0D; infection (p=1.000), cervical cancer and persistent HR-HPV&#x0D; subclinical infection (p=0.430), persistent and clearance HR-HPV&#x0D; subclinical infection group (p=0.740).&#x0D; &#x0D; Conclusion: This study proves that normal cervix group has the&#x0D; highest level of retinol deposit sufficiency; however, it cannot&#x0D; be stated that cervical cancer group has less sufficiency level.&#x0D; Persistent HR-HPV subclinical infection group has the lowest&#x0D; level of retinol deposit (OR 11.33). There is no association&#x0D; between sufficient level of retinol deposit and clearance of&#x0D; HR-HPV.&#x0D; &#x0D; [Indones J Obstet Gynecol 2017; 5-1: 46-54]&#x0D; &#x0D; Keywords: cervical cancer, HR-HPV clearance, retinol deposit

Boosting the SNR by adding a receive-only endorectal monopole to an external antenna array for high-resolution, T2 -weighted imaging of early-stage cervical cancer with 7-T MRI.

The aim of this study was to investigate the signal-to-noise ratio (SNR) gain in early-stage cervical cancer at ultrahigh-field MRI (e.g. 7T) using a combination of multiple external antennas and a single endorectal antenna. In particular, we used an endorectal monopole antenna to increase the SNR in cervical magnetic resonance imaging (MRI). This should allow high-resolution, T2 -weighted imaging and magnetic resonance spectroscopy (MRS) for metabolic staging, which could facilitate the local tumor status assessment. In a prospective feasibility study, five healthy female volunteers and six patients with histologically proven stage IB1-IIB cervical cancer were scanned at 7T. We used seven external fractionated dipole antennas for transmit-receive (transceive) and an endorectally placed monopole antenna for reception only. A region of interest, containing both normal cervix and tumor tissue, was selected for the SNR measurement. Separated signal and noise measurements were obtained in the region of the cervix for each element and in the near field of the monopole antenna (radius<30mm) to calculate the SNR gain of the endorectal antenna in each patient. We obtained high-resolution, T2 -weighted images with a voxel size of 0.7×0.8×3.0mm3 . In four cases with optimal placement of the endorectal antenna (verified on the T2 -weighted images), a mean gain of 2.2 in SNR was obtained at the overall cervix and tumor tissue area. Within a radius of 30mm from the monopole antenna, a mean SNR gain of 3.7 was achieved in the four optimal cases. Overlap between the two different regions of the SNR calculations was around 24%. We have demonstrated that the use of an endorectal monopole antenna substantially increases the SNR of 7-T MRI at the cervical anatomy. Combined with the intrinsically high SNR of ultrahigh-field MRI, this gain may be employed to obtain metabolic information using MRS and to enhance spatial resolutions to assess tumor invasion.

The morphology and distribution of CD1a positive Langerhans cells in normal and squamous cell carcinoma of cervix

IntroductionLangerhans cells (LCs), a type of dendritic cells are the professional antigen presenting cells present in the mucosa surfaces. They play an important role in antitumor immune response. The present study aims to find out the morphology and distribution of CD1a positive LCs in normal and squamous cell carcinoma of cervix. MethodsTwenty two normal and eleven ectocervical specimens with squamous cell carcinoma were processed for immunohistochemistry and stained with monoclonal mouse anti-human CD1a (Dako, USA). The morphology of CD1a positive LCs was studied using Olympus BX43 microscope. Morphometric analysis was done using Cellsens imaging analysing software. ResultsThere was a statistically significant difference in the number of LCs between normal (8±2.76) and squamous cell carcinoma of cervix (5.36±2.88). In the region of lymphatic infiltration both in epithelium and lamina propria, there were more number of LCs and most of the cells lost their dendritic processes in squamous cell carcinoma. 31.77% of the cells had no dendritic processes. The difference in the mean diameters of LCs was statistically significant (p=0.005) between normal and squamous cell carcinoma of cervix. DiscussionFewer number of CD1a positive LCs and their loss of dendritic processes in the squamous cell carcinoma of cervix compared to normal cervix indicate that immune responses are suppressed in patients with cancer.

Prostate-specific Membrane Antigen (PSMA) Expression in the Neovasculature of Gynecologic Malignancies: Implications for PSMA-targeted Therapy.

The goal of the study was to examine expression of prostate-specific membrane antigen (PSMA) in neovasculature of gynecologic cancers, as PSMA-targeted therapy has showed a promise in treatment of advanced carcinomas. The study included cervical carcinoma (n=28), vulvar carcinoma (n=20), endometrial carcinoma (n=23), primary ovarian carcinoma (n=21), metastatic ovarian carcinoma (n=25), and normal cervix (n=12) as negative control. All cases were immunostained using anti-CD31 antibody to delineate capillary endothelial cells. In parallel, all cases were immunostained using anti-PSMA antibody. The PSMA staining was assessed in tumor capillaries and in normal tissues and scored as a percentage of CD31 staining. PSMA expression was found in the tumor neovasculature, and no significant expression was identified in vasculature of normal tissues. The extent of PSMA staining in tumor capillaries varied from high expression in ovarian and endometrial cancers, to medium expression in cervical squamous cell carcinomas, and low expression in cervical adenocarcinomas and vulvar cancers. All (100%) cases of primary ovarian carcinoma, ovarian carcinoma metastases, and primary endometrial carcinoma showed PSMA expression in tumor vasculature, which was diffuse in majority of cases. The expression of PSMA in ovarian cancer metastases was similar among different metastatic foci of the same tumor. Fifteen percent of cervical squamous cell carcinoma, 50% of cervical adenocarcinoma, and 75% of vulvar carcinomas showed no capillary expression of PSMA. In conclusion, PSMA is highly and specifically expressed in the neovasculature of ovarian, endometrial, and cervical squamous carcinoma, rendering it a potential therapeutic vascular target.

Proteomic identification of potential biomarkers for cervical squamous cell carcinoma and human papillomavirus infection.

It is known that high-risk human papillomavirus infection is the main etiological factor in cervical carcinogenesis. However, human papillomavirus screening is not sufficient for early diagnosis. In this study, we aimed to identify potential biomarkers common to cervical carcinoma and human papillomavirus infection by proteomics for human papillomavirus-based early diagnosis and prognosis. To this end, we collected 76 cases of fresh cervical tissues and 116 cases of paraffin-embedded tissue slices, diagnosed as cervical squamous cell carcinoma, cervical intraepithelial neoplasia II-III, or normal cervix from ethnic Uighur and Han women. Human papillomavirus infection by eight oncogenic human papillomavirus types was detected in tissue DNA samples using a quantitative polymerase chain reaction. The protein profile of cervical specimens from human papillomavirus 16-positive squamous cell carcinoma and human papillomavirus-negative normal controls was analyzed by proteomics and bioinformatics. The expression of candidate proteins was further determined by quantitative reverse transcriptase-polymerase chain reaction and immunohistochemistry. We identified 67 proteins that were differentially expressed in human papillomavirus 16-positive squamous cell carcinoma compared to normal cervix. The quantitative reverse transcriptase-polymerase chain reaction analysis verified the upregulation of ASAH1, PCBP2, DDX5, MCM5, TAGLN2, hnRNPA1, ENO1, TYPH, CYC, and MCM4 in squamous cell carcinoma compared to normal cervix ( p < 0.05). In addition, the transcription of PCBP2, MCM5, hnRNPA1, TYPH, and CYC was also significantly increased in cervical intraepithelial neoplasia II-III compared to normal cervix. Immunohistochemistry staining further confirmed the overexpression of PCBP2, hnRNPA1, ASAH1, and DDX5 in squamous cell carcinoma and cervical intraepithelial neoplasia II-III compared to normal controls ( p < 0.05). Our data suggest that the expression of ASAH1, PCBP2, DDX5, and hnRNPA1, and possibly MCM4, MCM5, CYC, ENO1, and TYPH, is upregulated during cervical carcinogenesis and potentially associated with human papillomavirus infection. Further validation studies of the profile will contribute to establishing auxiliary diagnostic markers for human papillomavirus-based cancer prognosis.

Gene promoter methylation and protein expression of BRMS1 in uterine cervix in relation to high-risk human papilloma virus infection and cancer.

Cervical cancer is strongly related to certain high-risk types of human papilloma virus infection. Breast cancer metastasis suppressor 1 (BRMS1) is a tumor suppressor gene, its expression being regulated by DNA promoter methylation in several types of cancers. This study aims to evaluate the methylation status of BRMS1 promoter in relation to high-risk types of human papilloma virus infection and the development of pre-cancerous lesions and describe the pattern of BRMS1 protein expression in normal, high-risk types of human papilloma virus-infected pre-cancerous and malignant cervical epithelium. We compared the methylation status of BRMS1 in cervical smears of 64 women with no infection by high-risk types of human papilloma virus to 70 women with proven high-risk types of human papilloma virus infection, using real-time methylation-specific polymerase chain reaction. The expression of BRMS1 protein was described by immunohistochemistry in biopsies from cervical cancer, pre-cancerous lesions, and normal cervices. Methylation of BRMS1 promoter was detected in 37.5% of women with no high-risk types of human papilloma virus infection and was less frequent in smears with high-risk types of human papilloma virus (11.4%) and in women with pathological histology (cervical intraepithelial neoplasia) (11.9%). Methylation was detected also in HeLa cervical cancer cells. Immunohistochemistry revealed nuclear BRMS1 protein staining in normal high-risk types of human papilloma virus-free cervix, in cervical intraepithelial neoplasias, and in malignant tissues, where staining was occasionally also cytoplasmic. In cancer, expression was stronger in the more differentiated cancer blasts. In conclusion, BRMS1 promoter methylation and aberrant protein expression seem to be related to high-risk types of human papilloma virus-induced carcinogenesis in uterine cervix and is worthy of further investigation.

OVCA1 expression and its correlation with the expression levels of cyclin D1 and p16 in cervical cancer and intraepithelial neoplasia.

The present study aimed to examine the associations between the protein and mRNA expression levels of ovarian cancer gene 1 (OVCA1), cyclin D1 and p16 and high-risk human papillomavirus (HR-HPV) infection in cervical lesions. The protein expression levels of OVCA1, cyclin D1 and p16 in 66 cases of cervical cancer, 64 cases of cervical intraepithelial neoplasia (CIN) and 34 normal cervix tissues were detected using immunohistochemistry. The mRNA expression levels of OVCA1, cyclin D1 and p16 in cervical cancer and normal cervix cells were detected using real-time polymerase chain reaction. The results revealed that the protein expression levels of OVCA1 increased gradually, whereas its mRNA expression levels decreased gradually, in the progression from normal cervix tissue to CIN and cervical cancer (P<0.01). In addition, significant differences in the protein expression levels of OVCA1 between low-and high-level CIN, as well as between the early and advanced stages of cervical cancer, were observed (P<0.05). No significant associations were detected between the protein and mRNA expression levels of OVCA1 and the pathological type of cervical cancer or the presence of lymph node metastasis (P>0.05). The expression levels of OVCA1 mRNA and protein were positively correlated with the levels of p16 expression (P<0.01). Significant differences were also observed in the OVCA1 protein and mRNA expression levels between the HR-HPV (+) and HR-HPV (−) groups (P<0.05). Therefore, aberrant expression of OVCA1 protein and mRNA may be important during the development of cervical lesions, particularly in the early stages. In addition, the mechanisms underlying the effects of OVCA1 during cervical cancer development may involve p16 and HPV, as the levels of OVCA1 in cervical lesions were correlated with abnormal expression of p16 and HR-HPV infection.

Architectural overlap between benign endocervix and pattern-A endocervical adenocarcinoma: Are all pattern-A tumors invasive?

Studies on the pattern-based classification for invasive endocervical adenocarcinoma showed that tumors with nondestructive invasion (pattern-A) have a 0% rate of nodal metastases. Our understanding of pattern-A tumors and their distinction from in-situ adenocarcinoma requires further study. Thirteen sections diagnosed independently as pattern-A adenocarcinoma by three gynecologic pathologists, and 14 sections of benign endocervix were selected. Three additional pathologists (reviewers) evaluated a digital image from each section and classified it as pattern-A or benign based on architecture only. To blind the interpretation to cytologic features, nuclei and cytoplasm were obscured using morphometric software (Zen 2011, Carl Zeiss Microscopy, Germany). 13/27 cases (48%; 8 pattern-A, 5 benign) were correctly classified by all reviewers; 19/27 (70%; 10 pattern-A, 9 benign) were correctly classified by ≥2 reviewers. 3/13 pattern-A cases (23%) were interpreted as benign by ≥2 reviewers. Conversely, 5/14 benign cervices (36%) were misinterpreted as pattern-A by ≥2 reviewers. The number of glands per 20× field was higher in pattern-A cases with high reviewer agreement (p=0.004). An abnormal architecture is seen in many pattern-A adenocarcinomas in support of their invasive nature; some, however, have architecture that overlaps with that of benign endocervix thus may actually represent in-situ lesions. Likewise, normal cervix can be architecturally complex and mirror patterns that pathologists would classify as pattern-A if malignant cytologic features were present. Based on this overlap and the nil risk of nodal spread, an emphasis on the non-destructive, rather than the invasive, nature of pattern-A adenocarcinoma is recommended.

Gene expression analysis combined with functional genomics approach identifies ITIH5 as tumor suppressor gene in cervical carcinogenesis.

Progression from human papillomavirus-induced premalignant cervical intraepithelial neoplasia (CIN) to cervical cancer (CC) is driven by genetic and epigenetic events. Our microarray-based expression study has previously shown that inter-α-trypsin-inhibitor heavy chain 5 (ITIH5) mRNA levels in CCs were significantly lower than in high-grade precursor lesions (CIN3s). Therefore, we aimed to analyze in depth ITIH5 expression during cervical carcinogenesis in biopsy material and cell culture. Moreover, functional analyses were performed by ectopic expression of ITIH5 in different cell lines. We were able to confirm the validity of our microarray differential expression data by qPCR, demonstrating a clear ITIH5 downregulation in CC as compared with CIN2/3 or normal cervix. ITIH5 protein loss, evaluated by immunohistochemistry, was evident in 81% of CCs, whereas ITIH5 showed weak to moderate cytoplasmic staining in 91% of CIN2/3 cases. In addition, ITIH5 was strongly reduced or absent in seven CC cell lines and in three immortalized keratinocyte cell lines. Moreover, ITIH5 mRNA loss was associated with ITIH5 promoter methylation. ITIH5 expression could be restored in CC cell lines by pharmacological induction of DNA demethylation and histone acetylation. Functionally, ITIH5 overexpression significantly suppressed proliferation of SW756 cells and further resulted in a significant reduction of colony formation and cell migration in both CaSki and SW756 tumor models, but had no effect on invasion. Remarkably, ITIH5 overexpression did not influence the phenotype of HeLa cells. Taken together, ITIH5 gene silencing is a frequent event during disease progression, thereby providing evidence for a tumor suppressive role in cervical carcinogenesis.

Expressions of Immune Negative Regulator FoxP3+Treg and PD-L1 Protein in the Immune Microenvironment of Cervical Lesion.

Objective To explore the expression patterns of immune negative regulator FoxP3+Treg and PD-L1 protein in cervical carcinoma and its precancerous lesions. Methods The expression patterns of FoxP3+Treg and PD-L1 protein in 45 cases of cervical lesions tissues of the cervix and 20 cases of normal cervix tissues by using immunohistochemistry (IHC). Results Compared with the normal cervix,the expressions of FoxP3+Treg (H=43.211,P=0.000) and PD-L1 protein (t=213.00,P=0.001) were significantly increased in cervical lesions. Compared with the low-grade squamous cell carcinoma,the invasiveness of FoxP3+Treg was increased in high-grade squamous cell carcinoma (H=28.307,P=0.000),along with increased expression of PD-L1 protein (t=239.000,P=0.028). The FoxP3+Treg expression was positively correlated with PD-L1 protein expression in abnormally differentiated cells (rs=0.364,P=0.003). Conclusion Along with the progression of cervical cancer,the FoxP3+Treg amount increases in the local microenvironment,along with enhanced PD-L1 protein expression in abnormally differentiated cells.

Risk of high-grade precancerous lesions and invasive cancers in high-risk HPV-positive women with normal cervix or CIN 1 at baseline-A population-based cohort study.

Infection with high-risk human papillomavirus (HR-HPV) is transient and clears on its own in majority of the women. Only a few women who have persistent infection may finally develop cervical intraepithelial neoplasia (CIN) or cervical cancer in later years. The risk of progression in the HR-HPV-positive women with normal cervix or low-grade lesion on colposcopy and histopathology at baseline is less studied. We performed a longitudinal study on 650 HR-HPV-positive women with colposcopy and/or histopathology-proved normal or CIN1 diagnosis at baseline to assess the cumulative risk of development of high-grade CIN. After a mean follow-up of 2.1 person years of observation (PYO) (range 0.1-5.1), the cumulative incidence of CIN2+ (6.4%; 3.0/100 PYO) was significantly higher in women who had persistent HR-HPV infection compared to those who cleared the infection (adjusted HR 6.28; 95% CI 2.87-13.73). The risk of viral persistence in women aged 50-60 years was two times higher compared to women aged 40-49 years and three times higher compared to women aged 30-39 years. The probability of having persistent infection increased progressively with higher viral load at baseline (adjusted HR 3.29, 95% CI 2.21-4.90 for RLU ≥100; adjusted HR 2.69, 95% CI 1.71-4.22 for RLU 10-100). Women with increasing viral load at follow-up had four times higher risk of developing CIN2 or worse lesions as compared to those with decreasing load (20.9% vs 4.8%; p < 0.001). In the context of developing countries where cytology or genotyping triaging is not feasible, colposcopy referral of HR-HPV-positive women with advancing age, viral persistence, and increasing viral load may be considered.

γH2Ax Expression as a Potential Biomarker Differentiating between Low and High Grade Cervical Squamous Intraepithelial Lesions (SIL) and High Risk HPV Related SIL.

BackgroundγH2AX is a protein biomarker for double-stranded DNA breakage; its expression was studied in cervical squamous intraepithelial lesions and carcinomas.MethodsImmunostaining for phospho-γH2AX was performed in sections from histologically confirmed cervical SIL and carcinomas, as well as from normal cervices used as controls. In total, 275 cases were included in the study: 112 low grade SIL (LGSIL), 99 high grade SIL (HGSIL), 24 squamous cell carcinoma (SCC), 12 adenocarcinoma and 28 cervical specimens with no essential lesions. Correlation of histological grading, high risk vs. low risk HPV virus presence, activated vs. non-activated status (by high risk HPV mRNA expression) and γH2AX expression in both basal and surface segments of the squamous epithelium was performed.ResultsGradual increase of both basal and surface γH2AX expression was noted up from normal cervices to LGSIL harboring a low risk HPV type, to LGSIL harboring a high risk virus at a non-activated state (p<0.05). Thereafter, both basal and surface γH2AX expression dropped in LGSIL harboring a high risk virus at an activated state and in HGSIL.ConclusionsγH2AX could serve as a potential biomarker discriminating between LGSIL and HGSIL, as well as between LGSIL harboring high risk HPV at an activated state.

Negative immune factors might predominate local tumor immune status and promote carcinogenesis in cervical carcinoma

BackgroundThe disequilibrium of local immune microenvironment is an essential element during tumorigenesis.MethodBy conducting real-time polymerase chain reaction, we identified the mRNA level of immune factors, FoxP3 (forkhead box protein P3), CCL22/CCR4 (chemokine (C-C motif) ligand 22/CC chemokine receptor 4), OX40L/OX40 (tumor necrosis factor superfamily member 4/tumor necrosis factor receptor superfamily member 4) and Smad3 (SMAD family member 3) in neoplastic foci and its periphery tissues from 30 cases of squamous cervical carcinoma and 20 cases of normal cervix.ResultThe FoxP3, CCL22 and CCR4 mRNA level in local immune microenvironment of normal cervix was lower than that in cervical cancer. While OX40L, OX40 and Smad3 mRNA level profile in normal cervix was higher than that in cervical cancer. Beyond individual effect, the pairwise positive correlations were demonstrated among the mRNA level of FoxP3, CCL22 and CCR4. The mRNA level of OX40 negatively correlated with CCL22, but positively correlated with Smad3. Moreover, the mRNA level of FoxP3 and CCL22 was increased while Smad3 was decreased in cervical tissue with HPV (human papilloma virus) infection.ConclusionOur data yields insight into the roles of these immune factors in cervical carcinogenesis. It may therefore be that, in microenvironment of cervical squamous cell carcinoma, along with the context of HPV infection, negative immune regulators FoxP3, CCL22 and CCR4 might overwhelm positive immune factors OX40L, OX40 and Smad3, giving rise to an immunosuppressive status and promote the progression of cervical carcinogenesis.Trial registrationNot applicable.