Cervical Neoplasia Research Articles

Влияние терапии цервикальных неоплазий на иммунологические и апоптотические показатели

Aim: Evaluation of immune response factors in the treatment of mild cervical neoplasia against the background of human papillomavirus in patients of childbearing age. Design: Prospective study. Materials and methods. The study included a survey of 210 patients of fertile age. Among them were 150 patients with human papillomavirus (HPV) and cervical intraepithelial neoplasia, established by histological examination. Patients underwent Pap test, extended colposcopy, determination of HPV type and viral load. Patients with suspected pathology of the cervical zone underwent multifocal biopsy of the cervix. When analyzing changes in the cervix zone, the levels of interferon-γ, interleukin-18, caspase 3 and 9 were evaluated. Results. Among patients with cervical neoplasia of the first degree, the concentration of caspase 3, caspase 9 and interferon-γ at the first stage of the examination was higher (p < 0.05) than the values in the control group (among conditionally healthy women without viral load), while interleukin-18 was significantly lower (p < 0.05). In subgroups I and II, after 3 and 12 months, the concentrations of the studied parameters significantly differed from those in the control group and values before treatment (p < 0.05), while, after 12 months, the values of caspase 3 and caspase 9 in the subgroup where therapy was carried out (subgroup II) were significantly lower than in subgroup I. Also, among the patients, there was a statistical difference in the values of the cytokine series, however, the level of interleukin-18 in subgroup II after 10 days and 12 months was significantly higher than in subgroup I. fluctuations in indicators made it possible to justify the use of immunocorrective therapy for the treatment of a mild neoplastic process of the cervical zone. Conclusion. The results of the study allow optimizing approaches to the treatment of women of childbearing age with mild cervical neoplasia on the background of HPV. Keywords: genotyping, caspase-3, caspase-9, apoptosis, cervical neoplasia, polymerase reaction, interleukin-18, tumor necrosis factor-α, viral load.

Факторы формирования неопластических процессов в шейке матки

Aim: Review the literature data on the pathogenetic aspects of the formation of squamous intraepithelial lesions and the role of immunological mechanisms in the formation of this group of cervical diseases. Key Points. Cervical cancer (CC) is a multi-stage process that is often preceded by human papillomavirus (HPV)-associated intraepithelial lesions. The establishment of causal interactions between the virus and malignancy, as well as the study of epidemiological data, leads to the formation of models of cervical carcinogenesis: infection, persistence, progression to the precancerous stage, and finally, cervical cancer. Despite the clear and obvious path of malignancy, as well as the availability of generally available diagnostic methods in the arsenal of a practicing obstetrician-gynecologist, the assessment and prediction of the further development of cervical pathologies in patients of active reproductive age remains difficult. In general, cervical cancer can be considered an excellent model for understanding the staging of malignancy processes, which will be primarily associated with the impact of human papillomavirus infection and changes in the immunological aspects of endo-and exo-cervix. Conclusion. An analysis of sources published in the Cochrane Librare, Google Scolare and PubMed systems was carried out. In recent years, the understanding of human papillomavirus-associated tumor interaction with the host immune system has improved, so the development of new approaches targeting immune checkpoints has sparked interest in the use of immunotherapy in cervical cancer. CC is curable if detected at an early stage. However, reliable diagnostic and prognostic markers related to the physiological and pathological regulation of cervical cancer are currently not available. Therefore, the approach to the treatment of cervical cancer has remained unchanged for several decades, and new diagnostic strategies are currently required, due to the interaction of the immune system and the virus. Keywords: genotyping, apoptosis, cervical neoplasia, polymerase reaction, interleukin-18, tumor necrosis factor alpha, viral load.

ОСОБЕННОСТИ ЛОКАЛЬНОГО ИММУННОГО ОТВЕТА И ИММУНОГИСТОХИМИЧЕСКИЕ МАРКЕРЫ ПРОГНОЗА ПРИ РАКЕ ШЕЙКИ МАТКИ

According to modern data, cervical cancer (CC) occupies one of the leading places in the structure of malignant neoplasms in women of the Republic of Belarus and the Russian Federation; it is also the fourth most common cancer among women worldwide and the seventh in the general statistics of the incidence of human malignant tumors. Currently, there is a tendency towards an increase in the incidence of cervical cancer among young women, and therefore the problem of diagnosing and evaluating the prognosis of this tumor pathology is becoming increasingly important. The process of carcinogenesis in the cervix has a complex multifactorial nature and includes many biochemical mechanisms. To evaluate them various immunohistochemical markers are used. In order to determine the biological potential of squamous cell carcinoma of the cervix, the researchers have evaluated the role of the expression of p53, Ki67, cyclin D1 and CD45 proteins; these markers have also been supposed to be used as a tool for early cancer diagnosis. However, the data on the role of local immunity in assessing the invasive and metastatic potential of a malignant neoplasm are still extremely scarce. The article presents current literature data on the prognostic role of the expression of immunohistochemical markers in cervical cancer and the features of local immunity in cervical neoplasia.

Analysis of the Prevalence and Quantification of viral load of different human papillomavirus types in Turkish women population

Persistent infection with high-risk (HR) Human Papilloma Virus (HPV) genotypes has been shown to play a significant role in the development of cervical intraepithelial neoplasia (CIN) and CC (cervical cancer). The present study aimed to determine the distribution and quantification of viral load of HPV genotypes in numerous genital samples obtained from women undergoing routine gynaecological care in different regions of Turkey. HPV typing was done by HPV QUANT-21 Quantitative RT-PCR Kit®, which is intended for the specific identification and quantification of low-risk (HPV 6, 11, 44) and high-risk (HPV 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 82) from female subjects in Turkey. From the total of 5975 samples, 2777 were positive for at least one HPV genotype, with an overall frequency of 46.4%. 1695 patients were positive for only one or more LR-HPV genotypes (61%) and 812 patients were positive for one or more HR-HPV genotypes (29%). The frequency of LR-HPV genotypes was 31.4%, while the frequency of HR-HPV genotypes was 118.8%. Our tecnology had a positive advantage to calculate the concentration of each genotypes. Although genotype 52 ranked fifth in frequency, it showed the highest mean concentration, with a value of 5.38 log (copies/sample). The presence and genotype of viruses before HPV vaccination have also gained importance. The data obtained would provide guidance for prevention strategies, mainly of vaccination. We decided to add a new estimate to the effectiveness of currently available HPV vaccines and the development of screening programs to prevent and decrease the incidence of CC in Turkey. Further studies would be planned to measure and define the high infection level that can lead to the development of cervical neoplasia. Using this tecnology could give us a clinical desicion to degree the cytological changes.

Evaluation of co-testing with cytology and human papillomavirus testing in cervical screening

Cervical screening is increasingly switching to human papillomavirus (HPV) testing. In many settings, the switch has involved one or several co-tests (testing using both cytology and HPV) in the screening guidelines, to ensure safety. When Sweden switched to HPV testing in 2015 the guidelines included a co-test at age 41.To evaluate the effect of co-testing, we identified all 208,701 women resident in Sweden who in 2019 were 40–42 years old and thus eligible for co-testing. All cervical samples, the results of the test and of the subsequent biopsies were identified in the Swedish National Cervical Screening Registry. Out of the 10,643 women with co-testing in screening, there were 197 women with a subsequent biopsy with high-grade cervical neoplasia or worse (CIN2+). Among these 197 women, 189 had a screening test positive for both HPV and cytology, 6 women were HPV+/Cyt- and 2 women were HPV-/Cyt+. There were 7115 women with a co-test outside of the screening program. Among these, 325 women had a CIN2+ in histopathology, 290 were double positive, 13 women were cyt+/HPV-, and 11 women each were HPV+/cyt- and HPV-/Cyt-. In summary, the additional yield of CIN2+ with co-testing was 2 cases per 10,643 women as compared with 195/10,643 CIN2+ cases detected with HPV screening alone. However, for cervical samples taken outside the screening program (e.g. taken on a clinical indication) there was an increased yield (314 CIN2+ cases detected with co-testing as compared to 301 cases with HPV screening).

Extended human papillomavirus genotype distribution in cervical intraepithelial neoplasia and cancer: Analysis of 40 352 cases from a large academic gynecologic center in China.

Our aim was to conduct a large epidemiologic analysis of the distribution of human papilloma virus(HPV) genotypes associated with cervical neoplasias and cancers at a major Chinese gynecologic center. The pathologic database was searched for cervical histopathologic diagnoses with prior HPV genotyping from liquid cervical cytology specimens obtained ≤6 months before biopsy. HPV testing was performed by using the Tellgenplex HPV27 or YanengBio HPV23 genotyping assays. A total of 40 352 cases meeting study criteria were identified. High risk human papillomavirus (hrHPV) was detected in 94.1% of squamous cancers compared to in only 83.3% of cervical adenocarcinomas. The prevalence of multiple HPV infections was highest in cervical intraepithelial neoplasia 1 (CIN1) (33.8%) and decreased with increasing severity of squamous lesions. The distribution of HPV genotypes was similar between CIN1 and histopathologic-negative cases. HPV16 was one of the three most common hrHPV genotypes before all histopathologic abnormalities, ranging from 72.0% for cervical cancers, 38.7% for CIN2/3/AIS, 13.1% for CIN1, and 9.1% for biopsy-negative cases. HPV16 and HPV18 accounted for over 87.2% of detected hrHPV genotypes for all glandular intraepithelial neoplastic lesions and cancers, whereas squamous lesions did not show this pattern. 80.3% of cervical cancers were associated with genotypes covered by HPV16/18 vaccines and 89.6% with genotypes covered by 9-valent vaccination.

Cervical neoplasia in relation to socioeconomic and demographic factors - a nationwide cohort study (2002-2018).

Cervical cancer is a major cause of mortality and morbidity. We aimed to estimate the association between sociodemographic factors and cervical neoplasia. In this Swedish nationwide open cohort study, 4 120 557 women aged ≥15 years at baseline were included between January 1, 2002 and December 31, 2018. The two outcomes were cervical cancer and carcinoma in situ identified in the Swedish Cancer Register. Sociodemographic factors (age, education level, family income level, region of residency, country of origin) were the main predictors. Incidence rates per 10 000 person-years were calculated. Cox regression was used to estimate hazard ratios. Sensitivity analyses were conducted, including parity, urogenital infections, alcohol- and drug-use disorders, and chronic obstructive pulmonary disease (used as a proxy for tobacco abuse). In 38.9million person-years of follow-up, 5781 (incidence rate: 1.5, 95% confidence interval [CI] 1.4-1.5) and 62 249 (incidence rate 16.9, 95% CI 15.9-16.1) women were diagnosed with cervical cancer and carcinoma in situ, respectively. Women from Eastern Europe had a hazard ratio of 1.18 (95% CI 1.05-1.33) for cervical cancer compared with Swedish-born women, while women from non-Western regions were inversely associated with cervical cancer and carcinoma in situ. Women with a low education level had a hazard ratio of 1.37 (95% CI 1.29-1.45) for cervical cancer compared with women with a high education level. Women from the Middle East and Africa living in Sweden seem to suffer less from cervical neoplasia, whereas women with low education and women from Eastern Europe seem to suffer more from cervical cancer.

The analysis of anal cytology positivity in women with pathological cervical cytology

Objective: Cervical cytology (CC) is a routine screening method used to reduce cervical cancer. Although anal cancer and cervical cancer have similar etiological factors the opinion about the anal cancer screening program is unclear. We aimed to determine the features of women with abnormal anal cytology (AC) who had screened via CC for cervical neoplasia.
 
 Methods: Two hundred and five females' CC results were investigated. The patients with normal CC were excluded, finally 87 participants were included. The demographics, medical, sexual, and reproductive features, CC and AC results were analyzed.
 
 Results: The study group had a mean age of 40.77 ± 9.50 years. AC was pathological in six patients (6.9%). Four of these (66.7%) were high-grade squamous intraepithelial lesions (HSIL) and two (33.3%) were low-grade squamous intraepithelial lesions (LSIL). The CC results of these patients were all HSIL, all of them were human papillomavirus (HPV) positive, with the most common type being 16 (83.3%). 
 
 Conclusion: Women with HSIL in CC (especially with concomitant HPV) may be riskier for AC positivity. The others are most likely to have negative AC results. The use of AC for early diagnosis of risky anal intraepithelial lesions (such as a screening tool) may be considered for this group of patients.

Assessing the risk of cervical neoplasia in the post-HPV vaccination era.

This review is based on the recent EUROGIN scientific session: "Assessing risk of cervical cancer in the post-vaccination era," which addressed the demands of cervical intraepithelial neoplasia (CIN)/squamous intraepithelial lesion (SIL) triage now that the prevalence of vaccine-targeted oncogenic high-risk (hr) human papillomaviruses (HPVs) is decreasing. Change in the prevalence distribution of oncogenic HPV types that follows national HPV vaccination programs is setting the stage for loss of positive predictive value of conventional but possibly also new triage modalities. Understanding the contribution of the latter, most notably hypermethylation of cellular and viral genes in a new setting where most oncogenic HPV types are no longer present, requires studies on their performance in vaccinated women with CIN/SIL that are associated with nonvaccine HPV types. Lessons learned from this research may highlight the potential of cervical cells for risk prediction of all women's cancers.

Colposcopic evaluation of cervix in patients with bacterial vaginosis

Background and aim Bacterial vaginosis (BV) is a clinical syndrome emerging from changing of the bacteria Lactobacillus species that produce hydrogen peroxide in the vagina with many population of anaerobic bacteria. A lot of researchers have agreed with the relation between BV and cervical intra-epithelial lesions. However, other researchers have not accepted this relation between cervical neoplasia and BV. The study aim was to evaluate the abnormal colposcopic findings associated with BV and to study the relation between BV and cervical lesions. Patients and methods This study is a cross-sectional observational one that included 246 women diagnosed as having BV using Amsle’s criteria. All participants underwent proper history taking, general and abdominal examination, local vaginal examination, and colposcopic examination. Finally, Swede score was applied to abnormal colposcopic findings. Results By naked-eye examination, there were 163 patients with normal findings and 83 patients had abnormal cervical appearance, colposcopic examination of patients revealed 14 patients with unsatisfactory findings, and 196 patients had normal findings and 36 patients had abnormal cervix. The abnormal colposcopic findings were significantly associated with urban (55.6%) than rural women (44.4%) (P=0.039), educated (72.2%) than noneducated women (27.8%) (P=0.033), nonemployed (72.2%) than employed women (27.8%) (P=0.011), and women using oral contraceptive pills (OCP) (38.9%) than other contraceptive methods (P=0.024). Conclusion Most cervical lesions associated with BV in our study were of low grade and although use of colposcopy allowed early detection and management of cervical precancerous lesions associated with BV.

Association Between Hormonal Contraceptives and Cervical Neoplasia in Local Population of Khyber Pakhtunkhwa

Background: Hormonal contraception and the risk of developing cervical cancer is a contentious topic. These debates may reduce the popularity of hormonal birth control. Objectives: The goal of this study is to assess the prevalence of Cervical Smear abnormalities among Peshawar's multiethnic population in relation to the usage of hormonal contraceptives. Methods: This was a Case Control Study conducted on the females visiting DHQ Teaching Hospital Swabi, RHSC (A) KTH, HMC, LRH and FWC’s in district Peshawar who were persistent long-term users for more than 5 years of any type of hormonal contraception. The study was conducted during the period from 1st August, 2021 to 30th January, 2022. We made two groups, one who had never used any method of contraception and the other persistent users of hormonal contraception. After approval from ethical committee and an informed consent of the client, enrollment in the study was made. Results: A total of 300 patients were included in this study. 150 were cases whereas 150 were controls. Mean and SDs For Age 30+4.6 28+4.92. In the hormonal group, for type of contraceptives used, 98 (65.33%) patients used injectable, 40 (26.66%) patients used OCP whereas 12 (8%) patients used implants. As per frequencies and percentages for duration of use of contraceptives in the hormonal group, 120 (80%) were using different types of contraceptives for less than or equal to seven years whereas 30 (20%) were found using contraceptives for more than seven years As per frequencies and percentages for smear results in both groups, only 3 (2%) patients were found having atypical smear results who using injectable contraceptives for more than seven years however in the control group only 02 (1.33%) patient were having atypical smear results. Conclusion: Our research did not identify any correlation between using birth control and having an abnormal Pap smear. Additional prospective research is needed. Keywords: Pap smear, Abnormal, and Oral Contraceptive Pill (OCP).

Association Between Human Leukocyte Antigen Polymorphism and Human Papillomavirus Infection in Brazilian Women.

Background Human papillomavirus (HPV) infection is a necessary cause for cervical cancer, but a majority of genital HPV infections clear spontaneously. Human leukocyte antigen (HLA) polymorphism influences immune response and genetic susceptibility, and its association with cervical cancer was extensively investigated, but few reports focused on HPV infection. Methods We performed molecular typing of HLA-A, HLA-B, HLA-DQB1, and HLA-DRB1 genes as well as of HPV in 1226 women enrolled in the Ludwig-McGill cohort study and investigated the influence on cumulative HPV positivity. HPV types were grouped according to Alphapapillomavirus subgenera that exhibit similar tissue tropism and biological behavior concerning cancer risk. The associations between HLA polymorphisms and HPV infections were estimated using unconditional logistic regression analysis adjusted for age and race. Results HLA-B*08 and HLA-DRB1*15:01 were negatively associated with HPV positivity, and similar effects were observed for HPV subgenus 2 only, which includes HPV16. HLA-DRB1*08:07 was associated with overall HPV infection and subgenus 2 positivity. The haplotypes HLA-B*08-DRB1*03:01-DQB1*02:01 and HLA-DRB1*08:07-DQB1*04:02 were negatively and positively associated with cumulative HPV positivity, respectively. Conclusions Our data suggest that HLA class I and II polymorphism can influence HPV natural infection.

Prognostic value of systemic inflammation response index in patients with persistent human papilloma virus infection.

Persistent human papilloma virus (HPV) infection is a risk factor for the progression of cervical neoplasia into invasive carcinoma. Many inflammatory markers obtaining from hemogram parameters as platelets, monocytes, lymphocytes, and neutrophils or their ratios are still under investigation in recent decades, especially in the oncology era. Indeed, there have not been enough data about the relationship between these parameters and cervical cancer in the literature. Our primary aim was to investigate the possible relationship between the persistent HPV, which is one of the significant risk factors of cervical cancer, and these inflammatory markers. Further, we can add an easy follow-up parameter in women with persistent HPV infection. The study included patients between 30-65 years old, tested positive for HPV, and afterward had an HPV control test between January 2015 and June 2020. The study included 114 HPV DNA-positive patients. The mean age was 43 (standard deviation 8.7), and 41 of them (36%) had persistent HPV, but the remaining 73 (64%) did not. The baseline neutrophil/lymphocyte ratio (NLR) value was 2.1, platelet/lymphocyte ratio (PLR) was 133, monocyte/lymphocyte ratio (MLR) was 0.28, and systemic inflammation response index (SIRI) was 0.9. All the parameters were significantly higher in the persistent HPV group compared to the non-persistent group. Patients who had 0.65 and under this had a significantly lower risk of persistent HPV. Persistent HPV disease can be predicted with an elevated SIRI, NLR, and other hematologic parameters. So, we can closely follow up with these patients with different algorithms to prevent cervical cancer.

Nomogram for predicted probability of cervical cancer and its precursor lesions using miRNA in cervical mucus, HPV genotype and age

Cervical cancer is the fourth most common cancer in women worldwide. Although cytology or HPV testing is available for screening, these techniques have their drawbacks and optimal screening methods are still being developed. Here, we sought to determine whether aberrant expression of miRNAs in cervical mucus could be an ancillary test for cervical neoplasms. The presence of miRNAs in 583 and 126 patients (validation and external cohorts) was determined by real-time RT-PCR. Performance of a combination with five miRNAs (miR-126-3p, -451a -144-3p, -20b-5p and -155-5p) was estimated by ROC curve analysis. Predicted probability (PP) was estimated by nomograms comprising -ΔCt values of the miRNAs, HPV genotype and age. A combination of five miRNAs showed a maximum AUC of 0.956 (95% CI: 0.933–0.980) for discriminating cancer. Low PP scores were associated with good prognosis over the 2-year observation period (p < 0.05). Accuracy for identifying cancer and cervical intraepithelial neoplasia (CIN) 3 + by nomogram was 0.983 and 0.966, respectively. PP was constant with different storage conditions of materials. We conclude that nomograms using miRNAs in mucus, HPV genotype and age could be useful as ancillary screening tests for cervical neoplasia.

Tandem mass tag-based quantitative proteomic analysis of cervical cancer.

Cervical cancer is a common cancer in women caused by high-risk human papillomavirus (Hr-HPV). Many potential biomarkers have been proposed for precancerous lesions and cancer diagnosis and some of these markers studied for prognosis. This study determined potential biomarkers for cervical cancer diagnosis in regard to HPV genotype by using isobaric labeling quantitative proteomics. in the current study, there were 75 formalin fixed paraffin embedded (FFPE) uterine cervical samples that used to determine the 14 HPV genotypes and the viral load of each genotype was determined. The tandem mass tag (TMT) proteomic work was performed on four FFPE samples of cervical cancer and four FFPE of control samples. The validation of biomarkers from cervical proteome were evaluated using Immunohistochemistry (IHC) testing. The most frequent HPV genotype among all other genotypes was HPV 16. There were 2753 proteins quantified by TMT and 336 of these proteins had significant differential abundances. KPNA2, MCM2, COL1A1, and DCN were selected based on functional enrichment analysis and validated by Immunohistochemistry (IHC) testing. The staining of IHC confirmed the upregulation of KPNA2 and MCM2 expression in cervical neoplasia and the downregulation of DCN and COL1A1 in some cervical cancer group subjects. The KPNA2 marker was compared to other previously reported biomarkers and is a putative biomarker to be validated in further studies, specifically the relationship with HPV load.

Effectiveness of Oxidized Regenerated Cellulose-Based Haemostat (Surgicel®) for Monopolar Electrosurgical Conization and Its Risk Factors for Postoperative Rebleeding: An Observational Study

Background: Prompt detection and treatment of high-grade cervical intraepithelial neoplasia (CIN), namely, CIN3, are important for preventing malignant transformation. In most cases, such treatment is performed through conization, but minimally invasive procedures must also be considered. Therefore, we present a surgical method using a combination of monopolar electrosurgical conization (MESC) and oxidized regenerated cellulose-based haemostat (ORCH) (Surgicel®) and discuss its merits and risk factors. Methods: The medical records of 274 patients treated at our hospital from January 1, 2015, to December 31, 2021, were retrospectively reviewed. To detect risk factors for postoperative problems, we considered 2 indicators: postoperative recurrence and haemostatic suture for severe postoperative bleeding. We also focused on the following 10 representative factors for predicting significant risk for postoperative problems: (1) advanced age, (2) high BMI, (3) nulliparity, (4) smoking history, (5) long operation time, (6) intraoperative suture, (7) positive margin, (8) invasive squamous cell carcinoma (SCC), (9) no dysplasia, and (10) cervical glandular neoplasia (CGN). Results: Among the 274 patients, 19 instances of postoperative recurrence and 8 instances of haemostatic suture were detected. In a multivariate analysis of the 10 factors, we detected no significant impact except for “no dysplasia” on both indicators. This result indicated that our method had no specific risk and could provide a certain extent of usability. Conclusions: The combination of MESC and ORCH for the treatment of CIN3 could be performed effectively and safely with minimal intraoperative haemostatic suturing of the cut surface.

Histopathological study of malignant lesion of Cervix in a tertiary care hospital of Nepal

Background and objectives: Like in other developing countries, cancer of cervix is one of the leading malignancies in women in Nepal. In Nepal cervical pap-smear screening is not done my most of due to lack of awareness. The objective of this study is to find the prevalence of abnormal cervical epithelial lesions.&#x0D; Materials and methods: This was a retrospective study of 140 hysterectomy specimen reported from the Department of Pathology, Birat Medical College in Biratnagar. All tissue blocks of cervical tissues were retrieved, stained by Haematoxyline and Eosin (H &amp; E) stain and re-examined. All the case was reviewed by consultant pathologist of Birat medical college.&#x0D; Results: In the present study out of 140 cases 08 ie. 5.71% have cervical epithelial neoplasia I. 1.42% cases is of CINII, 0.71 is of CINIII and 2.85% is squamous cell carcinoma.&#x0D; Conclusion: This study has demonstrated that 10.69% of the examined cases were abnormal. The squamous cell carcinoma is leading cancer in our study ie. 2.85%. In developing countries if patient is detected with abnormal pap smears then hysterectomy is indicated.

Combined Liquid Biopsy Methylation Analysis of CADM1 and MAL in Cervical Cancer Patients.

Simple SummaryIn cervical neoplasia, tissue methylation of CADM1 and MAL has been identified as a promising triage tool for HPV-positive women. Here, we performed an exploratory study on the potential of the methylated genes as combined markers for the analysis of cervical smears and peripheral blood samples in cervical cancer patients. The combination of both markers in liquid biopsy resulted in a sensitivity of 83.3% and specificity of 95.5% for discrimination of cervical cancer from healthy controls. The robustness of the assay was proven by a respectably high sensitivity in stage I-II patients.Cervical cancer is the fourth most common cancer in women, which is associated in >95% with a high-risk human papillomavirus (HPV) infection. Methylation of specific genes has been closely associated with the progress of cervical high-grade dysplastic lesions to invasive carcinomas. Therefore, DNA methylation has been proposed as a triage for women infected with high-risk HPV. Methylation analyses of cervical cancer tissue have shown that cell adhesion molecule 1 (CADM1) and myelin and lymphocyte protein (MAL) methylation are present in over 90% of all cervical high-grade neoplasias and invasive cervical cancers. Here, we established a liquid biopsy-based assay to detect MAL and CADM1 methylation in cell free (cf)DNA of cervical cancer. Methylation of the target gene was validated on bisulfite converted smear-DNA from cervical dysplasia patients and afterward applied to cfDNA using quantitative real-time PCR. In 52 smears, a combined analysis of CADM1 and/or MAL (CADM1/MAL) showed methylation in 86.5% of the cases. In cfDNA samples of 24 cervical cancer patients, CADM1/MAL methylation was detected in 83.3% of the cases. CADM1/MAL methylation was detected already in 81.8% of stage I-II patients showing the high sensitivity of this liquid biopsy assay. In combination with a specificity of 95.5% towards healthy donors (HD) and an area under the curve (AUC) of 0.872 in the receiver operating characteristic (ROC) analysis, CADM1/MAL cfDNA methylation detection might represent a novel and promising liquid biopsy marker in cervical cancer.

BV associated bacteria specifically BVAB 1 and BVAB 3 as biomarkers for HPV risk and progression of cervical neoplasia.

Background Bacterial vaginosis (BV) is associated with high-risk HPV (hrHPV) genotypes. There is a proposed bidirectional relationship between hrHPV and vaginal microbial diversity. This study investigated the association between BV associated bacteria in women co-infected with Human immunodeficiency virus (HIV) and hrHPV. Methods Stored cervical cytobrush samples were used for real time PCR detection of eight BV associated bacteria. Analysis of BV bacteria detected against HPV infection, socio-demographics and HIV data were conducted in R Statistical computing software of the R Core Team, 2020, version 3.6.3. Results A total of 190 samples were analysed. A. vaginae (p <0.001) BVAB 1 (p <0.001), BVAB 2 (p =0.428), BVAB 3 (p <0.001), Lactobacillus species (p =0.016) and S. sanguinegens (p =0.007) were associated with prevalent hrHPV. Increasing CIN severity was independently associated with detection of BVAB 1 OR 1.51(95% CI: 0.42-5.55), BVAB 3 OR 2.72(95% CI:0.90-8.55) and S. sanguinegens OR 1.02(95% CI:0.37-2.80). All HPV genotypes/groups, gravida <2, A. vaginae (p =0.002) and BVAB 1 (p =0.026) were significantly associated with HPV persistence. BVAB 3, p =0.010 and HPV 16 were significantly associated with HPV reinfection. Conclusion There is a significant association of A. vaginae, BVAB 1, BVAB 3, S. sanguinegens and Lactobacillus spp to prevalent hrHPV. BVAB 1, BVAB 3 and S. sanguinegens had an increased odds for increasing CIN severity. A vaginae, BVAB 1, gravida and all the HPV genotypes/groups were significantly associated with HPV persistence. Only BVAB 3 and HPV 16 were significantly associated with hrHPV reinfection at 1 year review. BVAB 1 and BVAB 3 are possible biomarkers for HPV infection and CIN progression.

Association of a genetic variant in the ATP-binding cassette sub-family B member 1 with risk of cervical cancer

BackgroundCervical cancer is one of the most prevalent cancers among women and is associated with high morbidity and mortality. Several factors are involved in the pathogenesis of cervical cancer including viral infections and/or genetic modifications that include single nucleotide polymorphisms (SNPs). MethodsWe aimed to investigate the association of genetic variants in ATP Binding Cassette Subfamily B Member 1 (ABCB1) (rs1128503) and Fc Gamma Receptor IIIa (FCGR3A) (rs396991) genes and susceptibility to cervical cancer in 263 individuals with or without this form of cancer. DNA samples were isolated and genotyped using a TaqMan based real-time (RT-) PCR method. ResultsThe rs1128503 was found to be associated with an increased risk of cervical cancer (recessive model: OR = 2.6, CI: 1.1–5.8, p = 0.02). whilst there was no association between the rs396991 and susceptibility to cervical cancer. ConclusionOur results showed that there is an association between the ABCB1 polymorphism and susceptibility for cervical cancer suggesting its potential as a risk stratification marker for cervical neoplasia.