Bosnian Journal of Basic Medical Sciences

The impact of Ki-67 index, squamous differentiation and several clinicopathologic parameters on the recurrence of low and intermediate-risk endometrial cancer

Endometrial endometrioid carcinoma (EEC) represents approximately 75–80% of endometrial carcinoma cases. Three hundred thirty-six patients with EEC followed-up in the authors’ medical center between and 2010–2018 were included in our study. Two hundred seventy-two low- and intermediate- EEC patients were identified using the European Society for Medical Oncology criteria and confirmed by histopathological examination. Recurrence was reported in 17 of these patients. The study group consisted of patients with relapse. A control group of 51 patients was formed at a ratio of 3:1 according to age, stage, and grade, similar to that in the study group. Of the 17 patients with recurrent disease, 13 patients (76.5%) were stage 1A, and 4 patients (23.5%) were stage 1B. No significant difference was found in age, stage, and grade between the case and control groups (p>0.05). Body mass index, parity, tumor size, lower uterine segment involvement, SqD, and Ki-67 index with p<0.25 in the univariate logistic regression analysis were included in the multivariate analysis. Ki-67 was statistically significant in multivariate analysis (p=0.018); however, there was no statistical significance in SqD and other parameters. Our data suggest that the Ki-67 index rather than SqD needs to be assessed for recurrence in patients with low- and intermediate-risk EEC.

Whole-exome sequencing reveals rare genetic variations in ovarian granulosa cell tumor

Ovarian granulosa cell tumor (OGCT) is a rare ovarian tumor that accounts for about 2-5% of all ovarian tumors. Despite the low grade of ovarian tumors, high and late recurrences are common in OGCT patients. Even though this tumor usually occurs in adult women with high estrogen levels, the cause of OGCT is still unknown. To screen genetic variants associated with OGCT, we collected normal and matched-tumor formalin-fixed paraffin-embedded (FFPE) from 11 OGCT patients and performed whole-exome sequencing (WES) using Illumina NovaSeq 6000. A total of 1,067,219 single nucleotide polymorphisms (SNPs) and 162,155 insertions/deletions (indels) were identified from 11 pairs of samples. Of these, we identified 44 tumor-specific SNPs in 22 genes and four tumor-specific indels in one gene that were common to 11 patients. We used three cancer databases (TCGA, COSMIC, and ICGC) to investigate genes associated with ovarian cancers. Nine genes (SEC22B, FEZ2, ANKRD36B, GYPA, MUC3A, PRSS3, NUTM2A, OR8U1, and KRTAP10-6) associated with ovarian cancers were found in all three databases. In addition, we identified seven rare variants with MAF ≤ 0.05 in two genes (PRSS3 and MUC3A). Of seven rare variants, five variants in MUC3A are potentially pathogenic. Furthermore, we conducted gene enrichment analysis of tumor-specific 417 genes in SNPs and 106 genes in indels using cytoscape and metascape. In GO analysis, these genes were highly enriched in “selective autophagy”, and “regulation of anoikis”. Taken together, we suggest that MUC3A is implicated in OGCT development, and MUC3A could be used as a potential biomarker for OGCT diagnosis.

Tissue-based metabolomics reveals potential biomarkers for cervical carcinoma and HPV infection

Aberrant metabolic regulation has been observed in human cancers, but the corresponding regulation in human papillomavirus (HPV) infection-associated cervical cancer is not well understood. Here, we explored potential biomarkers for the early prediction of cervical carcinoma based on the metabolic profile of uterine cervical tissue specimens that were positive for HPV16 infection. Fifty-two fresh cervical tissues were collected from women confirmed to have cervical squamous cell carcinoma (SCC; n = 21) or cervical intraepithelial neoplasia (CIN) stages II-III (n = 20). Eleven healthy women constituted the controls (NCs). Real-time polymerase chain reaction (PCR) was performed to detect HPV infection in the tissues. High-resolution magic angle spinning nuclear magnetic resonance was utilized for the analysis of the metabolic profile in the tissues. The expression of rate-limiting enzymes involved in key metabolic pathways was detected by reverse-transcription quantitative PCR. An independent immunohistochemical analysis was performed using 123 cases of paraffin-embedded cervical specimens. A profile of 17 small molecular metabolites that showed differential expression in HPV16-positive cervical SCC or CIN II-III compared with HPV-negative NCs was identified. According to the profile, the levels of α- and β-glucose decreased, those of lactate and low-density lipoproteins increased, and the expression of multiple amino acids was altered. Significantly increased transcript and protein levels of glycogen synthase kinase 3 beta (GSK3β) and glutamate decarboxylase 1 (GAD1) and decreased transcript and protein levels of pyruvate kinase M2 (PKM2) and carnitine palmitoyltransferase 1A (CPT1A) were observed in the patient group (p < 0.05). HPV infection and cervical carcinogenesis drive metabolic modifications that might be associated with the aberrant regulation of enzymes related to metabolic pathways.

Impact of metabolic syndrome on the risk of endometrial cancer and the role of lifestyle in prevention

Endometrial cancer is the second gynecological cancer with the highest global incidence. Among many associated risk factors, Among the risk factors, metabolic syndrome is an important and preventable one. It comprises a group of conditions that often occur together: central adiposity, hyperglycemia, arterial hypertension, and atherogenic dyslipidemia. This review aimed to describe the epidemiological and biological relationship between metabolic syndrome and endometrial cancer, focusing on the role of lifestyle in prevention. A literature search was carried out in the PubMed database. 4824 publications were screened, and 123 were included for this review. The association between metabolic syndrome and endometrial cancer has been described. Chronic adipose tissue inflammation and insulin resistance are involved in the development of obesity, particularly visceral adiposity. These changes promote the ideal environment for the development of endometrial cancer. Strategies based on lifestyle modifications may be effective for the prevention of metabolic syndrome and consequently endometrial cancer. Some of these modifications include adopting a diet rich in fruits, vegetables, whole grains, and legumes, depending to the accessibility of these foods for each region. Avoiding ultra-processed foods and increasing daily physical activity were also some suggested modifications. We propose that women be screened for metabolic syndrome to establish early treatment and to possibly prevent endometrial cancer. Clinical trials designed to prove the effect of lifestyle modifications on the prevention of endometrial cancer are needed.

Plasma exosome-derived fragile-site associated tumor suppressor is a powerful predictor of prognosis in patients with ovarian cancer

Objective: To investigate the levels of plasma exosome-derived fragile-site associated tumor suppressor (FATS) and evaluate its predictive ability in ovarian cancer (OC) patients. Patients and Methods: Exosome-rich fractions were isolated from the plasma of enrolled 90 patients with OC. The levels of plasma exosome-derived FATS were detected with ELISA. Results: The levels of exosome-derived FATS in OC patient were significantly lower than in healthy controls (P < 0.001). The levels of plasma exosome-derived FATS were obviously higher in OC patients with low grade (1/2), FIGO stages I/II than high grade (3/4), stages III/ IV disease (P = 0.003; P < 0.001). The levels of plasma exosome-derived FATS were significantly higher in OC patients with no lymph node metastasis, no ascites than those with lymph node metastasis, ascites (both P < 0.001). The levels of plasma exosome-derived FATS were obviously higher in OC patients with CA-125 less than 35U/ml than more than 35U/ml (P < 0.001). Among all enrolled OC patients, both 5-DFS and 5-OS were shorter in patients who had low plasma exosome-derived FATS levels than that high levels (both P < 0.001). The AUROC of plasma exosome-derived FATS were 0.85(95% CI: 0.76-0.91) for 5-DFS, 0.91(95% CI: 0.83-0.96) for 5-OS prediction in patients with OC, respectively. Conclusions: Plasma exosome-derived FATS levels in OC patient were significantly down-regulated. Low levels of plasma exosome-derived FATS had close relationship with FIGO stages I/II, low grade, ascites, higher levels of CA-125, lymph node metastasis and prognosis of OC patients. Our findings may provide a new strategy in treating OC.

HPV infection screening using surface plasmon resonance in women from Kunming (southwest China)

No study examined the frequency of human papillomavirus (HPV) genotypes by surface plasmon resonance (SPR) in southwest China. This was a cross-sectional survey (The Second Affiliated Hospital of Kunming Medical University, 10/2010 to 12/2011) in 150 patients who were hospitalized or volunteered for cervical cancer (CC) screening. A HPV typing kit was used to detect 24 types of HPV by the SPR technique. The HPV-positive rate was 34.8% in women with normal cytology and 92.9% in women with CC. The frequency of HPV16 increased from 9.4% for women with normal cytology to 28.9% for cervical intraepithelial neoplasia (CIN)1, 41.4% for CIN2, 54.1% for CIN3, and 71.4% for CC (p < 0.001). The frequency of HPV18 increased from 0% for women with normal cytology to 2.6% for CIN1, 3.4% for CIN2, 5.4% for CIN3, and 21.4% for CC (p = 0.03). HPV40 was only found in one patient with CC (p = 0.04). There was no relation between HPV genotype and women’s age. In Kunming (southwest China), the frequency of HPV infection was 74.0% among women who underwent CC screening. HPV16 and HPV18 were the two most frequent genotypes. SPR could be of value for the screening of HPV infection.

The role of glycogen synthase kinase 3 (GSK3) in cancer with emphasis on ovarian cancer development and progression: A comprehensive review

Glycogen synthase kinase 3 (GSK3) is a monomeric serine-threonine kinase discovered in 1980 in a rat skeletal muscle. It has been involved in various cellular processes including embryogenesis, immune response, inflammation, apoptosis, autophagy, wound healing, neurodegeneration and carcinogenesis. GSK3 exists in two different isoforms, GSK3α and GSK3β, both containing seven antiparallel beta-plates, a short linking part and an alpha helix, but coded by different genes and variously expressed in human tissues. In the current review, we comprehensively appraise the current literature on the role of GSK3 in various cancers with emphasis on ovarian carcinoma. Our findings indicate that the role of GSK3 in ovarian cancer development cannot be decisively determined as the currently available data support both prooncogenic and tumor-suppressive effects. Likewise, the clinical impact of GSK3 expression on ovarian cancer patients and its potential therapeutic implications are also limited. Further studies are needed to fully elucidate the pathophysiological and clinical implications of GSK3 activity in ovarian cancer.

Mechanism of OTUD5 in non‐small cell lung cancer cell proliferation, invasion, and migration

Ovarian tumor protease deubiquitinase 5 (OTUD5) has been discussed as a regulator of cancer development. Herein, the current study set out to explore the molecular mechanism of OTUD5 in non‐small cell lung cancer (NSCLC) cell proliferation, invasion, and migration. Firstly, the expression patterns of OTUD5, phosphatase and tensin homolog (PTEN), as well as microRNA (miR)-652-3p in cells were detected by qRT-PCR and Western blot. Cell viability, migration, and invasion were assessed with the help of cell-counting kit-8 and Transwell assays, in addition to the measurement of the ubiquitination and protein levels of PTEN. The binding relations between OTUD5 and PTEN, and miR-652-3p and OTUD5 were testified by co-immunoprecipitation or dual-luciferase assays. Cells were further treated with GSK2643943A (inhibitor of deubiquitinase) or miR-652-3p-inhibitor to explore the role of PTEN ubiquitination and miR-652-3p in NSCLC cells. OTUD5 and PTEN were both poorly-expressed, and miR-652-3p was highly-expressed in NSCLC cells. On the other hand, over-expression of OTUD5 suppressed NSCLC cell proliferation, invasion, and migration. OTUD5 deubiquitinated and stabilized PTEN, and miR-652-3p targeted and inhibited OTUD5 expression. Augmenting the ubiquitination levels of PTEN promoted NSCLC cell growth, whereas miR-652-3p inhibition promoted the tumor-suppressing effects of the OTUD5/ PTEN axis in NSCLC. Altogether, our findings highlighted that miR-652-3p restrained the role of OTUD5 in deubiquitinating PTEN to improve PTEN protein level, thereby promoting NSCLC cell proliferation, invasion, and migration.

Development and validation of a SEER-based prognostic nomogram for cervical cancer patients below the age of 45 years

This study aimed to establish a nomogram for the prognostic prediction of patients with early-onset cervical cancer (EOCC) in both overall survival (OS) and cancer-specific survival (CSS). The 10,079 patients diagnosed with EOCC between 2004 and 2015 were captured within the Surveillance, Epidemiology, and End Results (SEER) database and further were divided into training and validation sets randomly. The independent prognostic factors were identified in a retrospective study of 7,055 patients training sets randomly. Besides, the prognostic nomogram was developed using R software according to multivariable Cox regression analysis. Furthermore, the model was externally validated using the data of 3,024 patients diagnosed at different times enrolled in the SEER database. In training set, the C-indexes for OS and CSS prediction were 0.831 (95% confidence interval [CI]: 0.815-0.847) and 0.855(95%CI:0.839-0.871). The results of ROC indicated that nomograms possessed better predict performance compared with TNM-stage and SEER-stage. And the areas under the curve (AUC) of the nomogram for OS and CSS prediction in ROC analysis were 0.855(95%CI:0.847-0.864) and 0.782(95%CI:0.760-0.804), respectively. In addition, calibration curves presented perfect agreements between the nomogram-predicted and actual 1-, 3-, and 5-year in the validation cohort, in OS rate and CSS rate. This study established and validated a prognostic nomogram that provided an accurate prediction of 3-, 5-, and 10-year OS and CSS of EOCC patients, which contributed to clinicians to be useful for patients’ counseling and clinical trial designing.