Preliminary Study on the Expression of Testin, p16 and Ki-67 in the Cervical Intraepithelial Neoplasia

Cervical cancer is one of the most common malignant cancers in women worldwide. The 5-year survival rate is 65%; nevertheless, it depends on race, age, and clinical stage. In the oncogenesis of cervical cancer, persistent HPV infection plays a pivotal role. It disrupts the expression of key proteins as Ki-67, p16, involved in regulating the cell cycle. This study aimed to identify the potential role of testin in the diagnosis of cervical precancerous lesions (CIN). The study was performed on selected archival paraffin-embedded specimens of CIN1 (31), CIN2 (75), and CIN3 (123). Moderate positive correlation was observed between testin and Ki-67 as well as testin and p16 expression in all dysplastic lesions (r = 0.4209, r = 0.5681; p < 0.0001 for both). Statistical analysis showed stronger expression of the testin in dysplastic lesions vs. control group (p < 0.0001); moreover, expression was significantly higher in HSIL than LSIL group (p < 0.0024). In addition, a significantly stronger expression of testin was observed in CIN3 vs. CIN1 and CIN3 vs. CIN2. In our study, expression of Ki-67, p16, and testin increased gradually as the lesion progressed from LSIL to HSIL. The three markers complemented each other effectively, which may improve test sensitivity and specificity when used jointly.

Health-Related Quality of Life with Cervical Cancer and Precancer: A Cross-Sectional Study in Yunnan Province, China

Objectives Cervical cancer is the fourth most common cancer in women, with an estimated 600,000 new cases and 340,000 deaths worldwide in 2020. However, there remains limited understanding of the association between individual socioeconomic status (SES) and clinical characteristics and health-related quality of life (HRQoL) of women with cervical precancerous lesions and cancer worldwide, including in China. This study investigates socioeconomic and clinical variations in HRQoL of women with cervical precancerous lesions and cancer in Yunnan Province, China.MethodsThe present study employed a cross-sectional survey design. An EQ-5D-5L questionnaire was used to assess HRQoL of 400 patients with cervical precancerous lesions and cancer in Yunnan Province, China from 2019 to 2020. ANOVA and independent samples t-tests were performed to identify the independent variables associated with the EQ-5D-5L utility scores and VAS scores. Predictors of the utility scores were confirmed using a Tobit regression model.ResultsThe mean of cervical precancerous lesions and cancer patients’ EQ-5D-5L utility scores was 0.939 (SD, 0.104) and the mean of VAS scores was 80.84(SD, 16.551). Patients aged 40-59 years (β=-0.037, p=0.005), patients who were not aware of the human papillomavirus (HPV) vaccine (β=-0.032, p=0.004), and patients who underwent radical hysterectomy (β=-0.036, p=0.006) had significantly lower utility scores, whereas having high monthly household income (β=0.023, p=0.033) was significantly associated with higher EQ-5D-5L utility scores. Age (p=0.023) was statistically significant in differences in patients’ VAS scores. Anxiety/depression was the most frequently reported issue (35.75%) among participants. ConclusionsFuture cervical cancer prevention and treatment guidelines should focus on low-income women, women aged 40-59 years, and those lacking knowledge about cervical cancer prevention.

Stratified Mucin-Producing Intraepithelial Lesion (SMILE) of the Uterine Cervix: High-Risk HPV Genotype Predominance and p40 Immunophenotype

Stratified mucin-producing intraepithelial lesion (SMILE) is a rare high-grade cervical precancerous lesion designated a variant of adenocarcinoma in situ (AIS) in the WHO classification. We aimed to determine HPV genotypes, immunohistochemical phenotype and mucin presence in SMILE. Between 2010 and 2018, SMILE was diagnosed in 34 out of 6958 (0.5%) cervical biopsies, in 23 patients. Twenty-six tissue samples from twenty-one patients were available for further analysis, including 13 with SMILE alone, 12 with SIL and/or AIS and one with HSIL, AIS and endocervical adenocarcinoma. HPV genotyping was performed using the Seegene Anyplex II HPV 28 assay. Of the 26 samples, a single HPV genotype was identified in the majority of cases (n = 22), including 12/13 SMILEs associated with SIL/AIS. All but one were high-risk HPV genotypes (23/24; 96.8%). We identified seven different HPV genotypes, the most common being HPV16 (n = 10; 43.5%), HPV18 (n = 8, 34.8%) and HPV 31 (n = 5, 21.7%). All SMILEs showed a strong positive reaction to p16, CK7, CK19 and high Ki67 expression comparable to adjacent HSIL and/or AIS if present. SMILE showed variable mucin presence and p40-positive squamous differentiation suggesting phenotypic diversity in cervical precancerous lesions infected by single HPV.

The application of CRISPR/Cas9 system in cervical carcinogenesis

Integration of high-risk HPV genomes into cellular chromatin has been confirmed to promote cervical carcinogenesis, with HPV16 being the most prevalent high-risk type. Herein, we evaluated the therapeutic effect of the CRISPR/Cas9 system in cervical carcinogenesis, especially for cervical precancerous lesions. In cervical cancer/pre-cancer cell lines, we transfected the HPV16 E7 targeted CRISPR/Cas9, TALEN, ZFN plasmids, respectively. Compared to previous established ZFN and TALEN systems, CRISPR/Cas9 has shown comparable efficiency and specificity in inhibiting cell growth and colony formation and inducing apoptosis in cervical cancer/pre-cancer cell lines, which seemed to be more pronounced in the S12 cell line derived from the low-grade cervical lesion. Furthermore, in xenograft formation assays, CRISPR/Cas9 inhibited tumor formation of the S12 cell line in vivo and affected the corresponding protein expression. In the K14-HPV16 transgenic mice model of HPV-driven spontaneous cervical carcinogenesis, cervical application of CRISPR/Cas9 treatment caused mutations of the E7 gene and restored the expression of RB, E2F1, and CDK2, thereby reversing the cervical carcinogenesis phenotype. In this study, we have demonstrated that CRISPR/Cas9 targeting HPV16 E7 could effectively revert the HPV-related cervical carcinogenesis in vitro, as well as in K14-HPV16 transgenic mice, which has shown great potential in clinical treatment for cervical precancerous lesions.

Intercellular Adhesion Molecule-1 Gene Polymorphisms and Susceptibility to Cervical Cancer in the Northern Chinese Han Population

Many epidemiological studies have confirmed that ICAM-1 gene single-nucleotide polymorphisms (SNPs) are associated with susceptibility of various cancers, but there are relatively few studies on the relationship between ICAM-1 gene polymorphisms and the risk of cervical cancer. Therefore, we aimed to explore the potential role of ICAM-1 gene polymorphisms and the combined effect of SNPs in the pathogenesis of cervical cancer in Han women in northern China. This case–control group includes 488 cases of cervical cancer, 684 cases of cervical precancerous lesions, and 510 healthy females. Multiplex polymerase chain reaction (PCR) combined with the next-generation sequencing method was used for the determination of gene polymorphisms (rs5498, rs3093030, and rs281432). In our study, we divide cervical cancer into two subgroups: cervical squamous cell carcinoma (CSCC) group and cervical adenocarcinoma (CAC) group. We analyzed the alleles and genotypes of all research subjects using multivariate logistic regression analysis combined with 10,000 permutation tests. In addition, we also analyzed the distribution of haplotypes of the three SNPs in cervical cancer and cervical precancerous lesions. We found that the T allele and the dominant model of rs3093030 were associated with the susceptibility of cervical cancer (p = 0.042, p = 0.040, respectively). However, the significance disappeared after the Bonferroni correction for multiple testing (p &gt; 0.05). For rs5498, its mutant gene G, the codominant model, and the dominant model could reduce the risk of CAC (p = 0.009, p = 0.028, p = 0.011, respectively). Significant differences remained after Bonferroni correction (p &lt; 0.05, all). In addition, the frequency of haplotype “CTG” was significantly lower in the CAC group than in the controls. In conclusion, the study suggested that ICAM-1 gene polymorphisms may have a potential role in the pathogenesis of cervical cancer in the northern Chinese Han population.

Clinical application of the 2011 IFCPC colposcope terminology

Background Colposcopy offers an accurate way to the diagnose of cervical precancerous lesions. However, the diagnostic accuracy of colposcopy is unsatisfied. This study was to evaluate colposcopic accuracy according to the 2011 International Federation of Cervical Pathology and Colposcopy (IFCPC) terminology. Methods A retrospective cohort study was performed in 1,838 patients who underwent colposcopy in Shandong Qianfoshan Hospital, Cheeloo College of Medicine, Shandong University from October 2013 to April 2018. Using conization or cervical biopsy pathology as the gold standard, the agreement between colposcopic diagnosis and pathologic diagnosis was calculated, and correlations between variables were analyzed. Results As an authoritative and widely used terminology for colposcopy diagnosis, the 2011 IFCPC terminology has certain clinical practicality and diagnostic accuracy. However, some signs such as mosaic, punctation, sharp border, inner border sign and ridge sign had high specificity but unsatisfactory sensitivity, which limited the diagnostic value. Therefore, we discussed the Lugol’s staining, a very common sign in colposcopy, and analyzed the diagnostic significance of bright yellow staining in low-grade squamous intraepithelial lesion (LSIL) and mustard yellow staining in high-grade squamous intraepithelial lesion (HSIL). The results showed that mustard yellow may be a valuable indicator in the diagnosis of HSIL. Conclusion The 2011 IFCPC colposcope terminology has standardized interpretations of the colposcopic findings and improved the accuracy of colposcopy diagnosis. The aceto-white epithelium still has important diagnostic value; however, the value of a few signs is needed to be discussed and new signs are expected to be discovered. Although the significance of Lugol’s staining was diminishing, mustard yellow might be a valuable indicator for the diagnosis of HSIL.

Fluorescence colposcope with dye TMTP1-PEG4-ICG is comparable to the conventional colposcope with acetic acid and Lugol's iodine in identifying cervical precancerous lesions - A randomized controlled trial

Objective: To compare the diagnostic efficiency of fluorescence colposcope with dye TMTP1-PEG4-ICG versus conventional colposcope with acetic acid and Lugol’s iodine in identifying cervical precancerous lesions. Design and setting: Randomized controlled trial conducted at Colposcopy Center. Population: Women with abnormal cervical cancer screening results including cytology and/or HPV test. Methods: All participants were randomized to fluorescence colposcope group or conventional colposcope group. Patients of fluorescence colposcope group were applied dye TMTP1-PEG4-ICG to the cervix uteri. Patients of conventional colposcope group routinely administrated acetic acid and Lugol’s iodine to stain the cervix uteri. Main outcome measures: The colposcopists gave colposcope assessment impressions according to the cervical staining reactions and fluorescence signal-to-background ratio (SBR) calculation results. The diagnostic efficiency of fluorescence colposcope and conventional colposcope was calculated on a per-patient and per-site basis. Results: 195 women were successfully completed the study protocol and were randomized to fluorescence colposcope group (n=97) and conventional colposcope group (n=98). The accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of fluorescence colposcope on a per-patient basis were 61.9%, 66.7%, 61.0%, 23.8%, 90.9%, respectively. The above data corresponded to 73.9%, 69.6%, 74.2%, 15.5%, and 97.3% on a per-site basis in fluorescence colposcope group. In the conventional colposcope group, the above diagnostic indicators corresponded to 59.2%, 54.2%, 60.8%, 31.0%, 80.4%, respectively. Conclusions: The fluorescence colposcope with dye TMTP1-PEG4-ICG was comparable to the conventional colposcope with acetic acid and Lugol’s iodine, and exhibited better accuracy, sensitivity and excellent NPV on the basis of per cervical sites.