Comparison of microvascular density in cervical carcinoma in relation to predictive pathohistological parameters

Introduction/BackgroundIt has been concluded that the occurrence, development, invasion, and metastasis of tumors are closely linked to angiogenesis which is reflected by the tumor microvessel density. To analyze microvascular density...

VEGF expression and microvascular density in relation to high-risk-HPV infection in cervical carcinoma – An immunohistochemical study

Objective To explore the relationship between microvascular density(MVD) marked by CD105 monoclonal antibody (mAb) and stages of cervical cancer, clinical pathological factors. Methods From September 2005 to January 2009, cervical cancer specimens were stained by immunohistochemistry of CD105 monoclonal antibody marked by microvascular density in 30 cases of carcinoma in situ(stage 0), 15 cases of early invasive cervical cancer (stage ⅠA) and 35 cases of invasive cervical cancer (stage ⅠB-ⅡB). Furthermore, 50 cases with invasive cervical cancer were divided into group grade Ⅰ(n=5), group grade Ⅱ(n=35) and group grade Ⅲ (n=10) according to pathological grade of malignant tumor, and node-positive group (n=8) and node-negative group (n=42) according to lymph node metastasis. The expression of the vascular endothelial cell marked by CD105 monoclonal antibody in tumor interstitium of carcinoma in situ, early invasive cervical cancer, and invasive cervical cancer were detected by immunohistochemical SP method. Meanwhile, the correlation between the expression of microvascular density marked by CD105 monoclonal antibody in different stages of tumor development and clinicopathological factors was observed. Results The microvascular density marked by CD105 monoclonal antibody among carcinoma in situ(n=30, stage 0), early invasive cervical cancer (n=15, stage ⅠA) and invasive cervical cancer (n=30, stage ⅠB-ⅡB) were significant difference(P<0.05). But with the further test of L-S-D, microvascular density had no significant difference between carcinoma in situ and early invasive cervical cancer (P=0.528). Microvessel density in lymph node positive group was higher than that of lymph node negative group(P=0.035), and among group grade Ⅰ, group grade Ⅱ and group grade Ⅲ had no significant difference (P=0.059). Conclusion Microvascular density marked by CD105 is relative to clinical stages and prognosis of cervical cancer. It will give the therapy theory evidence of carcinoma in situ and early invasive cervical cancer. Key words: cervical cancer; angiogenesis; CD105; microvascular density (MVD)

Backgorund: Angiogenesis is associated with growth, dissemination and metastasis of tumours. Measurement of Microvascular Density (MVD) is a quantitative method of assessment of angiogenesis and would give a proportional co relate of the angiogenic process in tumours. The aim of this study is to measure the MVD by using CD34 staining in various phases of Chronic Myeloid Leukemia (CML) and type of CML (Granulocytic/Granulocytic Megakaryocytic) (G/GM) and to co-relate micro vascular densities with the grade of fibrosis. Bone marrow biopsy specimens of 30 CML patients and 20 non-CML (controls) cases that required bone marrow biopsy were subjected to CD34 staining and H&E staining. The mean MVD in CD34 slides was assessed by selecting hot spots and MVD was measured in these fields in high power (40 x magnification) and the mean MVD was calculated by taking the average of four hot spots per field. Grade of fibrosis and phase of CML, type (G/GM) were assessed in H&E slides. The controls were matched with respect to age and gender. Among 30 patients with CML, 21 were in chronic phase, five in accelerated and four in blast crisis. A normal distribution was obtained for MVD of both CML cases and controls using tests for normality. Comparison of mean MVD between CML and controls by student t-test showed a significant increase in MVD of CML cases (p = 0. 00026). However, no significant difference in MVD between the three phases viz, Chronic, accelerated and blast crisis phase (p = 0. 302) was obtained by using one way ANOVA. Comparison of Grade of fibrosis with MVD using independent t-test showed no significant difference in MVD between low (Grade1&2) and high grade (Grade 3&4) (p = . 805). No significant difference in MVD was obtained between G and GM types of CML using independent t-test (p = 0. 381). The study shows that there is a significant increase in MVD in CML cases than controls but no significant difference in MVD could be demonstrated between different phases of CML, histological types of CML and grades of fibrosis in CML.

Lymph node metastasis (LNM) is an important prognostic factor in cervical cancer (CC). In early stages, the risk of LNM is approximately 3.7 to 21.7%, and the 5-year overall survival decreases from 80% to 53% when metastatic disease is identified in the lymph nodes. Few reports have analyzed the relationship between miRNA expression and the presence of LNM. The aim of this study was to identify a subset of miRNAs related to LNM in early-stage CC patients. Formalin-fixed paraffin-embedded tissue blocks were collected from patients with early-stage CC treated by radical hysterectomy with lymphadenectomy. We analyzed samples from two groups of patients—one group with LNM and the other without LNM. Global miRNA expression was identified by microarray analysis, and cluster analysis was used to determine a subset of miRNAs associated with LNM. Microarray expression profiling identified a subset of 36 differentially expressed miRNAs in the two groups (fold change (FC) ≥ 1.5 and p < 0.01). We validated the expression of seven miRNAs; miR-487b, miR-29b-2-5p, and miR-195 were underexpressed, and miR-92b-5p, miR-483-5p, miR-4534, and miR-548ac were overexpressed according to the microarray experiments. This signature exhibited prognostic value for identifying early-stage CC patients with LNM. These findings may help detect LNM that cannot be observed in imaging studies.

This study aimed to detect the expression of eukaryotic translation initiation factor 3B (EIF3B) and investigate its correlation with tumor features and survival in cervical cancer patients. This study retrospectively reviewed 187 cervical cancer (squamous cell carcinoma) patients underwent tumor resection. Immunohistochemistry was performed to determine the expression of EIF3B in tissue samples. Besides, disease free survival (DFS) and overall survival (OS) were calculated. The median follow-up duration was 69 months, and the last follow-up date was 2017/12/31. EIF3B expression was higher in tumor tissue compared to paired adjacent tissue (45.5% vs. 32.1%, P= 0.015). Besides, EIF3B high expression was associated with higher Federation of Gynecology and Obstetrics (FIGO) Stage (P= 0.001) and presence of lymph node metastasis (P= 0.002). As to survival profiles, Kaplan-Meier curves disclosed that DFS (P< 0.001) and OS (P< 0.001) were both shorter in EIF3B high expression group compared to EIF3B low expression group. Multivariate Cox's regression analysis disclosed that EIF3B high expression, pathological grade III (vs I/II) and FIGO Stage III/IV (vs I/II) were independent predictive factors for unfavorable DFS as well as OS in cervical cancer patients (all P value < 0.05). EIF3B is overexpressed, and its high expression correlates with higher FIGO Stage, lymph node metastasis and unfavorable survival profiles in cervical cancer patients.

We retrospectively analysed the prognostic significance of changes in absolute neutrophil count (ANC), absolute lymphocyte count (ALC) and neutrophil/lymphocyte ratio (NLR) during treatment with definitive chemoradiotherapy (CRT) in 104 cervical cancer patients. The absolute white blood cell, ANC and ALC decrease during treatment, NLR increased throughout treatment and reached to a plateau at fifth week. The ANC and NLR after 3rd week of definitive CRT were significantly higher and ALC after 3rd week of treatment was significantly lower in patients with progressive disease compared patients with no evidence of disease. Patients in low-haematological risk (LHR) group had significantly higher number of patients with smaller tumour size, early stage disease and without lymph node metastasis. In multivariate analysis, high-haematological risk (HHR) group and lymph node metastasis were negative prognosticators of overall and disease-free survival (DFS). The presence of lymph node metastasis and HHR could serve as a predicative factor of poor prognosis for cervical cancer patients. IMPACT STATEMENT What is already known on this subject? The ANC and NLR after 3rd week of definitive CRT were significantly higher and ALC after 3rd week of treatment was significantly lower in patients with progressive disease compared patients with no evidence of disease. Patients in LHR group had significantly higher number of patients with smaller tumour size, early stage disease and without lymph node metastasis. Lymph node metastasis and HHR and were negative prognosticators of overall and disease-free survival (DFS). The presence of lymph node metastasis and HHR could serve as a predicative factor of poor prognosis for cervical cancer patients. What the results of this study add? Weekly changes in ANC, ALC, and NLR, especially after 3rd week of treatment are predictive factors of disease progression, not the high-risk features of disease. Furthermore, in HHR group more patients with extensive stage disease, larger tumour and lymph node metastasis were observed compared to LHR group. What the implications are of these findings for clinical practice and/or further research? The patients may be stratified according to risk factors. The treatment intensification maybe required for HHR patients compared to LHR patients. Since our findings are preliminary, further studies are required to support these findings.

Objective To investigate the expression of hypoxia inducible factor-1α (HIF-1α), matrix metalloprotein-9 (MMP-9) and microvascular density (MVD) in bladder cancer and its clinical significance. Methods 51 tissues diagnosed as bladder cancer cured in department of urology of our hospital from February 2013 to November 2015 were selected as research objects, and 30 cases of non-bladder cancer patients with normal bladder specimens used as negative control group. Immunohistochemical method was implemented to observe the expression of HIF-1α, MMP-9 and MVD in bladder tissues. Results The positive rate of MMP-9 and HIF-1α was significantly higher than that in normal bladder tissues [60.78% (31/51) vs. 0.00% (0/30), P=0.000; 70.58% (36/51) vs. 16.67% (5/30), P=0.000]; The score of the expression of HIF-1α, MMP-9 in bladder tissues were significantly higher than those in normal bladder tissues (4.31±1.03 vs. 1.12±0.07, P=0.000; 4.64±0.98 vs. 1.53±0.21, P=0.000). The expression of MVD in bladder cancer tissues were significantly higher than those in normal bladder tissues (38.06±9.04 vs. 19.17±5.19; t=11.946, P=0.000). The expressionof HIF-1α, MMP-9 and MVD in bladder cancer whose diameter more than 3 cm was significantly higher than that in tumor diameter whichless than 3 cm (P=0.025, 0.009, 0.011). The expression of HIF-1α, MMP-9 and MVD in invasive bladder cancerwas significantly higher than that of superficial bladder cancer tissue (P=0.000, 0.000, 0.035). Spearman correlation analysis results demonstrated that HIF-1α, MMP-9 expression showed a significant positive correlation with MVD expression levels (P=0.000, 0.001). Conclusion HIF-1α, MMP-9 and MVD in bladder cancer expression was significantly up-regulated and for assessment tumor size, clinical stage and pathological grade, which has good prospects for clinical application. Key words: Hypoxia inducible factor-1α; Matrix metalloprotein-9; Microvascular density; Bladder cancer; Expression; Clinical significance

Angiogenesis in Myelopoliferative Neoplasms

A8 Cervical cancer is one of the most common gynecologic malignancy in Hong Kong. The identification of pretreatment markers with predictive significance for the presence of lymph node metastasis in low stage of cervical cancer is clinically important. MicroRNAs (miRNAs) are noncoding RNA molecules of 21 to 24 nt that regulate the expression of target genes in a post-transcriptional manner. Recent evidence indicates that miRNAs play essential roles in tumorigenesis and cancer metastasis. This study was to evaluate the deregulated miRNAs associated with cervical cancer metastasis. A total of 20 cervical squamous cell carcinomas, in which 10 had pathologically confirmed lymph node metastasis and 10 no metastasis, were included in the study. RNAs were extracted from microdissected tumor tissue specimens. Real-time reverse transcription PCR technology-based miRNA array enabling quantitation of 365 human mature miRNAs, was used to profile global miRNA expression in cervical cancer. dChip software was used for biostatistical analysis of data. Global miRNA expression data obtained from all 20 arrays were normalized, and then the corresponding model-based expression index was calculated. Unsupervised hierarchical clustering analysis of data showed a distinct separation between cancers in the presence and absence of lymph node metastasis. Supervised hierarchical clustering analysis revealed that of 13 significantly differentially expressed miRNAs, 10 miRNAs including miR-137 (4.76-fold), miR-203 (3.82-fold), miR-594 (3.58-fold), miR-149 (3.05-fold), mir-365 (2.61-fold), miR-556 (2.43-fold), miR-33 (2.32-fold), miR-627 (2.12-fold), miR-20a (2.05-fold) and mir-653 (2.04-fold) were up-regulated and 3 miRNAs including miR-187 (-4.31), miR-4095p (-2.65-fold) and miR-615 (-2.1-fold) were down-regulated in the cervical squamous cell carcinomas with lymph node metastasis compared to that without metastasis. The results obtained from this preliminary study indicate that the deregulation of miRNAs appears to be involved in the cervical cancer progression, and miRNA expression signature can be of potential importance as predictive biomarker in cervical cancer metastasis. Further validation of such microRNA pattern by the testing set and an independent cohort of patients with cervical cancer as well as the functional study of these deregulated miRNAs are ongoing. Citation Information: Cancer Prev Res 2008;1(7 Suppl):A8.

IntroductionCyclin D1 had been associated with different clinical and pathological stages of cervical cancer; however, few studies had focused on its correlation with cervical cancer prognosis. Therefore, this study aimed to assess the expression of cyclin D1a and D1b in normal tissue, cervical cancer and cervical intraepithelial neoplasia and their effect on prognosis.MethodsExpression of cyclin D1a and D1b was detected by immunohistochemical staining in 78 cases of primary cervical cancer, 40 cases of cervical intraepithelial neoplasia, and 40 cases of normal cervical tissue.ResultsNo significant difference was observed in the expression of cyclin D1a between normal and cervical cancer tissues (P = 0.201); however, its expression was significantly higher in cervical cancer than in cervical intraepithelial neoplasia tissues (P = 0.000). Expression of cyclin D1b was higher in normal tissues than in cervical cancer tissues (P = 0.000). No significant difference was observed in the expression of cyclin D1a in cervical cancer tissues with respect to age, pathological type, clinical-stage, depth of tumor invasion, or presence of lymph node metastases (P = 0.111,0.119,0.539,0.084,0.539). COX survival analysis showed that lymph node metastasis might be an independent factor affecting postoperative recurrence (hazard risk [HR] = 0.240; 95% confidence interval [CI] = 0.968–30.156; P = 0.034).DiscussionCyclin D1a expression was associated with tumor tissue size and degree of differentiation. The expression of cyclin D1b in cervical cancer was associated with the presence of lymph node metastases. Cyclin D1a and D1b expression in cervical cancer tissue was significantly correlated. Cox survival analysis showed that the presence of lymph node metastases might serve as an independent factor affecting postoperative recurrence. The expression of cyclin D1a and D1b was not associated with cervical cancer prognosis.ConclusionAssessment of cyclin D1a and D1b expression in cervical cancer and cervical intraepithelial neoplasia revealed that cyclin D1 could not be used as a reference to assess cervical cancer patient prognosis.

The aim of our study was to identify predictive factors for lymph node metastases (LNM) in children and adolescents with papillary thyroid carcinoma (PTC) and their impact on survival. The authors conducted an Italian multicentric retrospective analysis on 132 pediatric patients (0-18years old) affected by PTC between 2000 and 2014. The investigated variables were demographic characteristics of the patients, clinicopathological features of PTCs, and persistence/recurrence of disease. The female/male ratio was 3.1:1. The median age was 14.3±3.5years (range 4-18years). Total thyroidectomy was performed in all the patients, followed by lymph node dissection in 87 patients (65.9%). Metastatic lymph node involvement was confirmed in 73 patients (55.3%): lateral compartment (LC) in 25 patients (34.2%), central compartment (CC) in 17 patients (23.3%), and both compartments in 31 patients (42.5%). Multifocality (P<.00), vascular invasion (P=.04), infiltration of the thyroid capsule (P<.00), minimal extrathyroidal extension (P<.00), diffuse sclerosing variant of PTC (P=.02), and presence of LNM in the LC (P<.00) were significantly associated with LNM in CC. Infiltration of the thyroid capsule (P<.00), massive extrathyroidal extension (P=.03), distant metastases (P=.02), PTC, not otherwise specified (P<.00), and presence of LNM in the CC (P<.00) were significantly associated with LNM in LC. Age, sex and size of PTC were not correlated with the presence of cervical LNM. Moreover, presence of LNM in CC increases the risk of persistence (P<.01) and recurrence (P<.02) of PTC in children and adolescents. Most predictors, unfortunately, are only identified post-operatively by histopathologic examination: Just a small part of them can be pre-operatively detected with a low-sensitivity neck ultrasonography. In PTC patients with pre-operative predictors, we suggest an accurate pre- and intra-operative evaluation of CC and/or LC to find suspicious lymph nodes. The presence of LNM in CC has an impact on disease/progression/relapse-free survival. We suggest performing RAI therapy and an accurate follow-up for pediatric patients with only post-operative predictors.

Background: cervical cancer is one of the most common malignancies in women worldwide and its management remains challenging and complex. As Cytochrome4Z1 (CYP4Z1) is overexpressed in many tumours, its expression in cervical cancer is unknown. Therefore, the present study aimed to evaluate CYP4Z1 expression in cervical cancers. Methods: CYP4Z1 expression was immunohistochemically assessed in 100 cases of cervical cancers along with ten normal cervix tissues, and the enzyme’s relationship to several clinicopathological features and survival was explored. Results: CYP4Z1 was strongly expressed in 55% of cervical cancer patients. Normal cervix samples were negative for CYP4Z1 expression. Importantly, this expression was significantly found in patients with the late stage of the disease, lymph node metastasis, and high tumour invasion (p < 0.05). Interestingly, CYP4Z1 expression was significantly correlated with shorter survival times of cervical cancer patients. Univariate analysis showed that CYP4Z1 expression, tumour stage, lymph node metastasis, and tumour invasion were significantly correlated with patient survival (p < 0.05). The multivariate analysis revealed that only CYP4Z1 expression and tumour stage were significantly correlated with patient survival (p < 0.05). Conclusions: CYP4Z1 expression is associated with cervical cancer patients’ survival and may serve as an independent predictor of poor prognosis in cervical cancer patients.

To determine risk factors for presence of lymph node or distant metastases in patients with follicular thyroid cancer (FTC) at the time of diagnosis and whether there is a relationship between the type of tumor invasion and metastases. FTC often presents distant metastases at the initial diagnosis. As distant metastases are independent prognostic factors in a patient's survival, determination of clinicopathologic characteristics for patients who are at higher risk for developing metastases is of greater clinical importance. The prognostic significance of gender (male vs. female), age (<or=40 years vs. <40 years), tumor size (<or=40 mm vs. >40 mm), number of lesions (uni- vs. multifocality), type of invasion (minimally invasive vs. widely invasive), and oncocytic changes (with vs. without) were analyzed in 207 patients, according to presence of lymph node and distant metastases at the time of initial surgery. According to the type of invasion, the carcinoma-specific survival and the disease-free survival of minimally invasive (MI) and widely invasive (WI) FTC were estimated and compared. None of the 127 patients with MI growth presented with lymph node metastases but 9.4% distant metastases. Overall risk factors for the presence of lymph node metastases at the initial diagnosis were multifocality (P = 0.02) and widely invasion (P = 0.0001) and for distant metastases age >45 years (P = 0.007), tumor size larger than 40 mm (P = 0.03) and widely invasion (P = 0.0001).WI-FTC patients show larger tumors (P = 0.0001), older age (P = 0.0001), and are presented more frequently in recurrent goiter disease (P = 0.0001). The estimated 10 years carcinoma-specific survival and disease-free survival for MI-tumors were significantly better than for WI-tumors (P = 0.0001). Total thyroidectomy is recommended in all patients with FTC because of early distant metastases. Patients with WI-FTC need a more aggressive surgical treatment because of higher tendency for lymph node metastases. MI-FTC has an excellent prognosis with no sign of lymph node metastases, which emphasizes a limited need for nodal surgery.

The clinical value of pretreatment serum concentrations of cytokeratin 19 fragments, measured by Cyfra 21-1, was compared with tissue polypeptide antigen (TPA) and squamous cell carcinoma antigen (SCC-Ag) in 78 patients with squamous cell cervical cancer. Serum levels were compared with tumor stage, size, lymph node status, parametrial involvement, and prognostic data. The clinical performance of the different tests was evaluated by their receiver operating characteristic (ROC) curves. Serum levels of all markers were related significantly to tumor stage and size. Elevated serum levels of these markers were not found to be predictive for the presence of lymph node metastases. In contrast, a positive relation was found between quantitative serum Cyfra 21-1, TPA, and SCC-Ag levels and the presence of either lymph node metastases or parametrial involvement (i.e., extracervical disease). An elevated, i.e. positive, serum Cyfra 21-1 level was related significantly to the presence of extracervical disease (P = 0.020). The clinical performance of each serum marker in predicting lymph node metastases or parametrial involvement appeared to be similar as expressed by their ROC curves. In the univariate analysis, Cyfra 21-1, TPA, and SCC-Ag showed prognostic value with respect to disease free interval and survival. Elevated serum levels were associated with a poor prognosis. However, after adjusting for tumor stage and size, none of these markers remained statistically significant. Cyfra 21-1 may be of additional value in assessing stage of disease, tumor size, and the presence of extracervical disease in patients with cervical cancer. Determining its value during follow-up warrants further study.