Croatian Medical Journal

Epidemiology of vulvar cancer in Croatia

To assess the incidence and mortality trends of invasive vulvar cancer in Croatia between 2001 and 2019/2020. The incidence data for the period 2001-2019 were obtained from the Croatian National Cancer Registry. The number of deaths from invasive vulvar cancer by age groups between 2001 and 2020 was obtained from the Croatian Bureau of Statistics. Joinpoint regression analysis was used to assess the trends and trend changes. Joinpoint regression analysis of vulvar cancer incidence rate showed a non-significant average annual percent increase (APC) of 0.8 (95% confidence interval [CI]=-0.3-2.0) during the whole period. There was also a non-significant increase in women under 60, with an average APC of 1.0 (CI = -1.6-3.7) during the whole period; similar results were obtained in women over 60 years of age (APC=0.9; CI=-0.3-2.1). The average annual percent increase in vulvar cancer mortality rate was 0.2% (CI = -1.0-1.5), with a similar trend in women over 60 years of age (APC=0.1; CI=-1.3-1.5). Mortality in women under 60 years of age was not assessed due to a very small number of deaths observed in the study period. In the studied period, the incidence of invasive vulvar cancer in Croatia was stable. Age-standardized rates (for all-ages, under 60, and over 60 years of age) increased, but the increase did not reach the level of statistical significance. The pattern in younger and older age groups was the same. The mortality rates over the last decade were stable.

Prognostic relevance of autophagy-related markers p62, LC3, and Beclin1 in ovarian cancer

To analyze the expression of autophagy markers p62, LC3, and Beclin1 in ovarian cancer tissue and evaluate the prognostic potential of these markers. The study enrolled 328 patients: 122 with epithelial ovarian carcinoma, 42 with atypical proliferative tumor, and 164 with benign epithelial ovarian tumor. The expression of p62, LC3, and Beclin1 was analyzed in central and invasive tumor segments with immunohistochemistry combined with tissue microarray. The expression levels of the analyzed markers were correlated with relevant histopathology parameters. The expression of all analyzed markers was most remarkable in epithelial ovarian carcinoma. There was a strong positive correlation between the expressions of p62 and LC3, while these two markers negatively correlated with Beclin1. High-grade serous carcinoma had higher p62 and LC3 levels, and lower Beclin1 levels than other tumor types. This expression profile was also observed in more advanced tumor stages. Prominent p62 and LC3 expression in combination with weak Beclin1 expression in high-grade serous carcinoma indicates potential for the application of autophagy inhibitors in patients with this tumor subtype.

Regulator of calcineurin 3 as a novel predictor of diagnosis and prognosis in pan-cancer

To assess diagnostic and prognostic value of regulator of calcineurin 3 (RCAN3) in various malignancies. RCAN3 expression levels were assessed across pan-cancer data sets including various molecular and immune subtypes. Receiver operating characteristic (ROC) and Kaplan-Meier curves were employed to determine the diagnostic and prognostic value of RCAN3 in pan-cancer, respectively. Enrichment analyses were used to identify RCAN3-associated terms and pathways. A special focus was placed on cervical squamous cell carcinoma and endocervical adenocarcinoma cervical cancer (CESC); we assessed the prognostic value of RCAN expression within distinct clinical subgroups and its effect on m6A modifications and immune infiltration. RCAN3 expression varied not only in different cancer types but also in different molecular and immune subtypes of cancers. RCAN3 displayed high accuracy in diagnosing and predicting cancers, and RCAN3 expression level was associated with the prognosis of certain cancers. CESC patients with a high RCAN3 level had a worse overall survival, disease-specific survival, and progression-free interval. RCAN3 expression was related to multiple m6A modifier genes and immune cells. In general, RCAN3 can serve as a novel biomarker for the diagnosis and prognosis in pan-cancer, especially in CESC. It may represent a promising molecular target for developing new treatments. However, our analysis is limited to bioinformatic predictions, and further biological experiments are necessary to verify our results.

Distribution of human papillomavirus genotypes in women with high-grade cervical intraepithelial lesions and cervical carcinoma and analysis of human papillomavirus-16 genomic variants

To analyze the distribution of high-risk human papillomavirus (HR-HPV) genotypes and the diversity of HPV-16 genomic variants in Croatian women with high-grade squamous intraepithelial lesions (HSIL) and cervical carcinoma. Tissue biopsy specimens were obtained from 324 women with histopathologically confirmed HSIL or cervical carcinoma, 5 women with low-grade SIL, and 49 women with negative histopathology. HR-HPV DNA was detected with Ampliquality HPV-type nucleic-acid hybridization assay, which identifies 29 different HPV genotypes. HPV-16 genomic variants were analyzed by an in-house sequencing. The most common HPV type in women with HSIL was HPV-16, detected in 127/219 (57.9%) specimens. HPV-16 was also the dominant type in squamous cell cervical carcinoma (46/69 or 66.7%) and in adenocarcinoma (18/36 or 50.0%). Out of 378 patients, 360 had HR-HPV (282 single infections and 79 multiple infections), 3 (0.8%) patients had low-risk HPV, and 15 (4%) tested negative. HPV-16 variants were determined in 130 HPV-16 positive specimens, including 74 HSIL and 46 carcinoma specimens. In HSIL specimens, 41 distinct variants were found, 98.6% belonging to the European branch and 1.4% belonging to the African branch. In cervical carcinoma specimens, 95% isolates grouped in 41 variants belonging to the European branch, one isolate (2.5%) belonged to the North American, and one (2.5%) to the Asian-American branch. HPV-16, mainly belonging to the European branch, was the most frequent HPV genotype in women from Croatia with histologically confirmed HSIL and cervical cancer.