Cervical Squamous Cell Research Articles

Clinicopathologic Significance of Atypical Glandular Cells in Cervical Cytology Smears: A five years Retrospective Study in a Tertiary Care Hospital

Introduction: Cervical cancer is the fourth most common cancer among women. Recently the incidence of cervicaladenocarcinoma has been increased significantly on the contrary to the decreasing incidence of cervical squamouscell carcinoma. Routine screening of cervical cancer by cervical smear cytology study to detect the pre-invasivelesions carries a significant role in reducing the incidence and mortality rate of cervical cancer.Objective: The aim of our study was to analyze the prevalence of Atypical Glandular Cells (AGCs) detected incervical smear screening, the follow up histopathological outcome and their clinical significance.Materials and Methods: This was a retrospective study done in College of Medicine and Sagore Dutta Hospital,Kolkata over a period of 5 years from June 2017 to June 2022. Patients detected with Atypical Glandular cells(AGCs) in cervical smears and having records of follow up histopathology reports were included in this study.The cases of AGCs in which the follow up histopathological reports were not available were excluded from thestudy.Result: The prevalence of atypical glandular cells were 0.57%. Out of the total 10,950 cervical smears examinedAGCs were found in 63 cases (0.57%). Out of 63 cases of AGCs subcategorization was done(according to TBS2015) which showed AGC not otherwise specified (AGC-NOS) in 46 cases (0.43%) and AGC Favoring Neoplasia(AGC-FN) were seen in 15 cases (0.13%). Both AGC -NOS and AGC-FN showed various pathology ranging fromreactive, metaplastic, benign to, in situ/Invasive carcinomas.Discussion: One of the major concern of AGC is that the cytological findings which characterize it have poorreproducibility between observers leading to interobserver variability. Due to the reported risk of premalignantand malignant lesions in AGCs it bear a great clinical significance and need for further histopathological correlation.
 Conclusion: Due to very low incidence of AGCs and the risk of malignancy associated with it multiple histologicalevaluation methods required for earlier diagnosis and management of the malignancies detected on histologicfollow up

Abstract 2442: Seminal fluid enhances cervical squamous carcinoma cell proliferation, epithelial mesenchymal transition (EMT), migration & invasion

Abstract Cervical cancer is the leading cause of cancer related deaths & the second most common cancer amongst South African women. Though there are various risk factors, the most common risk factor for cervical cancer development is sexual transmission & persistent infection with high-risk Human Papillomavirus (HPV). Although most HPV infections are cleared naturally in women, persistent infection with high-risk HPV types promotes neoplastic transformation & tumorigenesis. Furthermore, exposure of neoplastic cervical epithelial cells to inflammatory mediators may drive the progression of cervical cancer. Seminal fluid (SF), a biological fluid rich in inflammatory mediators & growth factors, has been implicated in cervical cancer progression. SF has been shown to induce the expression of proinflammatory mediators which promotes cell proliferation in culture & growth of explants in mice injected with HeLa cervical adenocarcinoma cells. Since majority of cervical cancer cases are squamous cell carcinoma, in this study, we examined the effect of SF on cell proliferation, EMT, cell migration & invasion employing SiHa and Me180 squamous cell carcinoma cell lines. Our results showed that SF significantly enhanced cell proliferation in both cell lines. Using Confocal microscopy and phalloidin staining we further showed that SF caused morphological changes and stress fibre formation. RT-qPCR showed that SF up regulated EMT transcription factors Snail, ZEB1 & TWIST mRNA expression. EMT induction was further confirmed by the increase of N-cadherin & a subsequent decrease in E-cadherin protein expression. Additionally, we showed that the induction of EMT transcription factors by SF occurs via the EGF/ERK/COX pathway. To investigate the effect of SF on migration and invasion, transwell migration assays were used. Results showed that SF significantly enhanced cell migration & invasion of the SiHa and Me180 cells. Our results show that SF plays a role in enhancing the migratory & invasive potential of squamous cell carcinoma cells & that this associates with increased expression of EMT transcription factors. These findings together implicate SF as a possible factor that may promote cervical cancer progression. Citation Format: Nonkululeko Mkwanazi, Virna D. Leaner, Arieh A. Katz. Seminal fluid enhances cervical squamous carcinoma cell proliferation, epithelial mesenchymal transition (EMT), migration & invasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2442.

Evaluating the utility of fluorine-18 fluorodeoxyglucose (18F-FDG)-positron emission tomography (PET)/ computed tomography (CT) scan in cancer of unknown primary.

3062 Background: Cancer of unknown primary (CUP) represents a heterogeneous group of metastatic tumours where standardized diagnostic work-up fails to identify the tissue of origin (TOO). Small studies, to date mainly focused on cervical lymph node squamous cell CUP patients, have shown 18F-FDG-PET/CT can change patient management and identify the TOO. We aimed to describe the Peter MacCallum Cancer Centre experience with 18F-FDG PET/CT in CUP with respect to detection of a TOO and its impact on management. A secondary aim was to compare the overall survival (OS) in patients where TOO is detected with those without TOO detection. Methods: Retrospective analysis of CUP patients treated between 2014-2020. Patients were identified from medical oncology clinics and PET/CT records. Information regarding demographics, clinicopathological details, CUP subtype as per ESMO guidelines, genomic analysis (If known), suspected TOO as per clinician pre- and post-FDG PET/CT, treatment details pre- and post FDG PET/CT and follow-up were collated from electronic medical records. Clinical details and genomic analysis were used to determine the clinically suspected TOO and compared against independent blinded nuclear medicine specialist FDG-PET/CT reads to determine sensitivity, specificity, accuracy and detection rate of TOO. Results: One hundred and forty-seven patients were identified of whom 65% had undergone molecular profiling. The median age at diagnosis was 61 years (range 20-84) and the median follow-up time was 69 months (range, 26–83). The predominant histological subtype was adenocarcinoma (54%). Eighteen percent of patients had a prior cancer history and 29% had a 1st degree relative with a history of cancer. Ninety-three percent were ECOG 0-1, and the dominant metastatic site was lymph nodes (35%). Eighty-one percent were classified as unfavourable CUP subtype as per ESMO guidelines. FDG PET/CT demonstrated a TOO detection rate of 34% with high specificity (98%) and moderate accuracy (78%). FDG PET/CT resulted in a change in management in 22% of patients and identified occult disease sites in 37% of patients. The median OS for all patients was 17.8 months. Median OS was not reached and 12.5 months for favourable and unfavourable CUP subtypes, respectively (p < 0.0001). Median OS when a potential TOO was identified on an FDG-PET/CT scan was 25.4 months compared with 9.1 months when a TOO remained elusive. (p < 0.0001). Multivariable analysis of survival adjusted for age and sex remained significant for FDG-PET identification of TOO (p = 0.004), favourable CUP (p < 0.001) and ECOG ≤ 1 (p < 0.001). Conclusions: 18F-FDG PET/CT plays a complementary role in CUP diagnostic work-up and was able to determine the likely TOO in a third of cases. OS is improved with TOO identification, demonstrating the value of access to a diagnostic PET/CT scan for CUP patients.

Role of APOA1 in the resistance to platinum-based chemotherapy in squamous cervical cancer

BackgroundTo investigate the mechanism by which apolipoprotein A1 (APOA1) enhances the resistance of cervical squamous carcinoma to platinum-based chemotherapy.MethodsTwo cervical squamous carcinoma cell lines (SiHa and Caski) overexpressing APOA1 were constructed, treated with carboplatin, and compared to normal control cells.ResultsIn both SiHa and Caski cell lines, the clone-forming ability of CBP-treated cells was lower than that of untreated cells, and the change in the number of clones of overexpressing cells was lower than that of normal control cells (p < 0.05), indicating that APOA1 overexpression enhanced chemoresistance. A screen for APOA1 downstream proteins affecting platinum-based chemoresistance using Tandem Mass Tag revealed 64 differentially expressed proteins in SiHa cells, which were subjected to Gene Ontology (annotation, Kyoto Encyclopedia of Genes and Genomes enrichment, subcellular localization, structural domain annotation and enrichment, clustering, and interaction network analyses. Sixty-four differentially expressed proteins matching cancer-relavent association terms were screened and parallel response monitoring identified 29 proteins as possibly involved in the mechanism of platinum-based chemoresistance.ConclusionsOur analysis suggested that the mechanism may involve numerous regulatory pathways, including promoting tumor growth via the p38 MAPK signaling pathway through STAT1, promoting tumor progression via the PI3K signaling pathway through CD81 and C3, and promoting resistance to platinum-based chemotherapy resistance through TOP2A. The present study aimed to preliminarily explore the function and mechanism of APOA1 in platinum-based chemoresistance in cervical cancer, and the detailed mechanism needs to be further studied.

Hinokitiol Dysregulates Metabolism of Carcinoma Cell Lines and Induces Downregulation of HPV16E6 and E7 Oncogenes and p21 Upregulation in HPV Positive Cell Lines

Background: Hinokitiol (β-thujaplicin), isolated from the wood of Chamaecyparis taiwanensis, has a wide variety of biological properties including anti-inflammatory, anti-microbial, and anti-tumor effects. Therefore, hinokitiol has become a frequent additive in oral and other healthcare products. Objectives: Our goal was to determine the anti-tumor activity of hinokitiol on human papillomavirus (HPV) positive (n = 3) and negative (n = 2) cell lines derived from cervical or head and neck squamous cell carcinoma (HNSCC) and keratinocyte cell lines (n = 3) transformed spontaneously or with HPV16E6 and E7 oncogenes. Methods: The cell-lines were exposed to hinokitiol at different concentrations (0–200 µM) for 24 h. Cell metabolism, proliferation, and the cell cycle distribution were assessed by MTT- and 3H-thymidine incorporation and flow cytometry. Expressions of p21 and on HPV16E6 and E7 oncogenes were assessed by qPCR. Results: In all carcinoma cell lines, hinokitiol treatment declined the metabolic activity irrespective of the HPV status. This decline was statistically significant, however, only in HPV-positive cell lines CaSki and UD-SCC-2 when exposed to hinokitiol concentrations at 100 and 200 µM, respectively (p &lt; 0.05). Immortalized cell lines, HMK and HPV-positive IHGK, were more sensitive as a similar metabolic effect was achieved at lower hinokitiol concentrations of 3.1, 6.25, and 50 µM, respectively. Hinokitiol blocked DNA synthesis of all carcinoma cell lines without evident association with HPV status. G1 cell cycle arrest and p21 upregulation was found in all cell lines after hinokitiol treatment at higher concentration. However, when the p21 results of all HPV-positive cells were pooled together, the increase in p21 expression was statistically significantly higher in HPV-positive than in HPV-negative cell lines (p = 0.03), but only at the highest hinokitiol concentration (200 µM). In HPV-positive cell lines hinokitiol declined the expression of HPV16E7 and E6 along the increase of p21 expression. The dose-dependent inverse correlation between p21 and E7 was statistically significant in SiHa cells (r = −0.975, p-value = 0.03) and borderline in UD-SCC-2 cells (r = −0.944, p-value = 0.06), in which p21 and E6 were also inversely correlated (r = −0.989). Conclusions: Our results indicate that hinokitiol might have potential in preventing the progress of immortalized cells toward malignancy and the growth of malignant lesions. Hinokitiol can also influence on the progression of HPV-associated lesions by downregulating the E6 and E7 expression.

Synthesis of 3-(2-(subsituted-(trifluoromethyl)phenylamino)acetyl)-2H-chromen-2-one derivatives as new anticancer agents

Under solvent free conditions and in presence of a base 3-(2-(subsituted-(trifluoromethyl)phenylamino)acetyl)-2H-chromen-2-one derivatives were synthesized by grinding technique. Structural investigations were carried out with IR studies, HRMS, 1HNMR and 13CNMR. The compounds were checked for their in vitro anticancer activities against three different human cancer cell lines viz human breast cancer cell line (MCF-7), human cervical cancer cell line (HeLa) and human oral squamous cell carcinoma (SCC-40) using SRB method. All the title compounds showed low toxicity towards non-malignant PBMC cells indicating their tumour selectivity. The compounds exhibited good in vitro anti-proliferative potency at lower concentrations against HeLa and MCF-7 cell lines and remain moderately active against SCC-40.

A case of cervical and retroperitoneal squamous cell carcinoma thought to be a G-CSF-producing tumor

A case of cervical and retroperitoneal squamous cell carcinoma thought to be a G-CSF-producing tumor

The prognostic role of radiotherapy and radiotherapy target in cervical lymph node metastatic squamous cell carcinoma with unknown primary: a retrospective study.

Aim to analyze the effect of radiotherapy for cervical lymph node metastatic carcinoma with unknown primary (CCUP) and compare the survival benefits between Comprehensive radiotherapy and Involved Field radiotherapy. The patients diagnosed with CCUP between 2009 and 2019 in our institution were analyzed retrospectively. The categorical variables were tested by χ2 test. Kaplan-Meier method was used for survival analysis. Log-rank test and Cox proportional hazards regression were performed with overall survival (OS) and disease-free survival (DFS) as the primary outcome variables. Of 139 patients, 64.7% (90/139) of them received radiotherapy. Of the 90 patients who underwent radiotherapy, 45.6% (41/90) received Involved Field radiotherapy and the rest 49 patients received Comprehensive radiotherapy. The median follow-up of 139 patients is 69months. The 1-year, 3-year, and 5-year OS rates are 87%, 62%, and 39%, respectively, and the DFS rates are 73%, 45%, and 29%, respectively. Multivariate analysis of 139 patients with CCUP shows that differentiation grade, N stage, radiotherapy, and the length of the largest lymph node (DmaxLN) are the independent prognostic factors for both OS and DFS. Subgroup analysis of 90 patients who received radiotherapy shows that the Comprehensive radiotherapy group has a better OS (P < 0.001) and DFS (P < 0.001) compared with Involved Field radiotherapy. Radiotherapy is the independent prognostic factor for CCUP. Comprehensive radiotherapy may be superior to Involved Field radiotherapy in survival benefits.

AdvanTIG-202: A phase 2 study investigating anti-TIGIT monoclonal antibody ociperlimab plus anti-PD-1 monoclonal antibody tislelizumab in patients with previously treated recurrent or metastatic cervical cancer.

TPS5595 Background: Women with recurrent/metastatic cervical cancer represent a poor prognostic group with high unmet clinical needs. T-cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibition motif domains (TIGIT) is a co-inhibitory, immune checkpoint receptor expressed on immune cells and upregulated on T-cells and natural killer cells in multiple solid tumors, inhibiting anticancer immune responses. Ociperlimab (BGB-A1217) is a novel, humanized, monoclonal antibody that binds TIGIT with high specificity and affinity, blocking the interaction with its ligands on tumor cells. Tislelizumab is an anti-PD-1 antibody engineered to minimize binding to FcγR on macrophages to abrogate antibody-dependent phagocytosis, a mechanism of T-cell clearance and potential resistance to anti-PD-1 therapy. Preclinical and clinical studies suggest that dual targeting with anti-TIGIT and anti-PD-1 antibodies produces synergistic immune cell activation and enhanced antitumor activity. Methods: AdvanTIG-202 is a Phase 2, randomized, multicenter, open-label study (NCT04693234). Approximately 167 pts with cervical squamous cell or adenosquamous carcinoma or adenocarcinoma, recruited from 100 centers, whose disease progressed on or after ≥ 1 prior line of chemotherapy for recurrent/metastatic disease will be included in this 2-Part study. In Part 1, approximately 80 pts will be randomized (1:1) to either ociperlimab 900 mg intravenously (IV) in combination with tislelizumab 200 mg IV every 3 weeks (Q3W) (Arm 1), or tislelizumab monotherapy 200 mg IV Q3W (Arm 2), until disease progression, unacceptable toxicity, or withdrawal of consent. In Part 2, Arm 1 will be expanded by approximately 87 additional pts whose tumors are evaluable for PD-L1 expression. The primary endpoint is overall response rate (ORR) (RECIST v1.1) assessed by Independent Review Committee (IRC) in Arm 1. Secondary endpoints are investigator-assessed ORR in Arm 2, IRC-assessed and investigator-assessed ORR in Arm 1, IRC-assessed and investigator-assessed duration of response, progression-free survival, time to response, disease control rate, clinical benefit rate and overall survival, cancer-specific health-related quality of life (HRQoL), safety, pharmacokinetics and immunogenicity in Arms 1 and 2. Exploratory endpoints are generic HRQoL and the association of biomarkers with patient prognosis, response or resistance. Clinical trial information: NCT04693234.

Snapshot of Clinical Implications of p16 Overexpression in Carcinogenesis

Molecular markers are needed to decide the treatment plans of certain cancer types when the histological and other clinical diagnoses are not sufficient to decide the tumor nodular metastasis (TNM) stage. The ubiquitous p16 gene is one of them gained popularity by fulfilling criteria to be a useful biomarker. Over expression of p16, to compensate the inactivity of another two tumor suppressor genes (TSG)s, pRb and p53 due to the integration of E7 and E6 high risk Human Papilloma viral oncoprotein respectively into the host keratinocytes is useful to consider the clinical impact of p16 biomarker in carcinogenesis. The p16 immunohistochemistry helps the diagnosis as well as prognosis of cervical and oropharyngeal squamous cell carcinoma, though there are ambiguities in the cutoff values of p16 positivity. There is also a re-emerging interest on clinical impact of p16 positivity in lung, breast, and colorectal cancer types. High risk HPV genotypes have been already established as the aetiological agents of cervical, other rare ano-genital and oropharyngeal (especially tonsils and base of the tongue) cancers. The HPV associated subset of head and neck cancers demonstrate a unique tumor biology, when compared with HPV non associated ones thus, most effective treatment planning including counselling is much needed to maximize the overall survival of HPV associated cancer patients, in the era of personalized precision medicine. In the shed of light, this communication glimpses on clinical implications of p16 overexpression in carcinogenesis not limiting to cervical and a subset of head and neck carcinomas (HNSCC).

The role of epidermal growth factor receptor as progression factor in cervical intraepithelial neoplasia and squamous cell carcinoma

Background: Cervical carcinoma is the most common cancer with a prevalence of 28.6% of all cancers in women in Indonesia and is one of the leading causes of death among gynecologic malignancies. Prevention and understanding are needed in terms of initiation and development of precancerous lesions into cervical cancer. About 90% of cervical cancers are squamous cell carcinomas that develop from intraepithelial neoplasia. Epidermal Growth Factor Receptor (EGFR) is an expression product of the proto-oncogene c-erbB-1 (HER-1), which plays a role in the proliferation, differentiation and acceleration of the transformation of malignant cells. This study aimed to prove the role of EGFR as a progression factor of cervical intraepithelial neoplasia and cervical squamous cell carcinoma.Methods: The study design was a cross-sectional analytic observational study, with a total sample of 36, which was taken from biopsy or surgery from patients with cervical intraepithelial neoplasia (CIN) and cervical invasive squamous cell carcinoma (SCC), whose tissues were examined at the Anatomical Pathology Laboratory, Faculty of Medicine, Universitas Udayana/Sanglah General Hospital. Histopathological diagnosis and determination of the type of malignancy were carried out by staining with Hematoxylin and Eosin (H &amp; E). The immunohistochemical stain evaluated EGFR expression.Results: The age range of CIN was 21-48 years and SSC 33-61 years, mean age 41.8 ± 10.55 years. A total of 16 (44.4%) showed CIN namely 9 (25%) low-grade CIN, 7 (19.4%) high-grade CIN, and 20 (55.6%) SCC. There was a significant difference in the expression of EGFR at low-grade CIN, high-grade CIN and SCC (p = 0.004). There was a significant difference in EGFR expression between low-grade CIN and high-grade CIN (p = 0.035) and between low-grade CIN and SCC (p = 0.003). There was no significant difference in EGFR expression between high-grade CIN and SCC (p = 0.441). Conclusion: EGFR has a role as a progression factor in CIN and SCC. There was no difference in the expression of EGFR between high-grade CIN and SCC, probably because in this study, carcinoma in situ was also included in high-grade CIN.

Novel hybrid endoscopy-assisted larynx-preserving esophagectomy for cervical esophageal cancer (with video)

Hybrid endoscopy-assisted larynx-preserving esophagectomy is developed for cervical esophageal squamous cell carcinoma encroaching or extending above the upper esophageal sphincter. First, a cervical incision was surgically performed followed by cervical lymph node dissection. Second, the margin of cervical esophageal squamous cell carcinoma was endoscopically identified with iodine staining and marked endoscopically followed by semi-circumferential or circumferential endoscopic full-thickness excision around the lumen of the esophagus. The distal margin was surgically resected and reconstruction was performed. Among six consecutive patients with cervical esophageal squamous cell carcinoma undergoing hybrid endoscopy-assisted larynx-preserving esophagectomy, proximal surgical margin was histologically negative in five patients. During a median follow-up period of 15.5months, all patients tolerated oral intake and were alive without evidence of recurrence. None of the patients experienced aspiration pneumonia, vocal disorder or postoperative anastomotic stricture. Hybrid endoscopy-assisted larynx-preserving esophagectomy could be a clinically feasible treatment for cervical esophageal squamous cell carcinoma providing accurate proximal resection margin with the benefit of laryngeal function preservation.

UCHL1 Promotes Lymph Node Metastasis in Scnec By Reducing PROX1 Ubiquitination

Study Objective Small cell neuroendocrine carcinomas of the cervix (SCNEC) are rare tumors with highly aggressive clinical behavior and extremely poor prognosis. Here, we investigate its differential genes and explore the roles of crucial gene for aggressive clinical behavior Design Gene expression microarrays and immunohistochemistry were used to screen out candidate genes in SCNEC. Cervical adenocarcinoma and squamous cell carcinoma samples were used as controls. The clinical significance of candidate genes was analyzed by SPSS 22.0, while biological functions were investigated in primary SCNEC cells using siRNA and overexpression. Co-expression network analysis and immunoprecipitation were used to explore molecular mechanisms. Setting Cervical adenocarcinoma and squamous cell carcinoma samples were used as controls. Patients or Participants Prognostic factors of SCNEC were analyzed in 53 SCNEC patients. Interventions None. Measurements and Main Results UCHL1, PROX1, CRMP5, and TM4SF1 were identified as preliminary candidates with different positively rates. According to the requirement of a >85% positive rate in sample size calculation, only UCHL1 was identified as a specific gene for SCNEC, which was positively associated with lymph node (LN) metastasis (p=0.003). LN metastasis was significantly related to a worse prognosis of SCNEC patients (P=0.021, HR =3.49, 95% CI 1.20-10.09). Scratch migration and transwell invasion assays indicated that the migration and invasion abilities of primary SCNEC tumor cells were significantly higher following UCHL1 overexpression and decreased by downregulation (P Conclusion UCHL1 is highly expressed in SCNEC and positively associated with lymph node metastasis. Mechanistically, UCHL1 promotes migration and invasion by reducing PROX1 ubiquitination.

The expression of p16 and galectin-3 in cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma (SCC) uterine cervix

Cervical cancer is the most prevalent cancer among women in India. The main cause of cervical cancer is persistent human papilloma viral (HPV) infection. HPV inactivates the pRb tumour suppressor protein; thus p16 expression, which is controlled by a negative feedback mechanism, is relatively increased. Galectin-3 is directly and indirectly connected to cancer cell activity and contributes to oncogenesis, angiogenesis, cancer progression and metastasis. Thus, the aim of this study was to study the expression of p16 and galectin-3 in Cervical Intraepithelial Neoplasia (CIN) and Squamous Cell Carcinoma (SCC) and to correlate p16 and galectin-3 expression. On hundred and eighteen newly-diagnosed untreated cases of CIN and SCC of uterine cervix were included in the study. Expression of p16 and galectin 3 was more pronounced in invasive SCC and High-grade Intraepithelial Lesion (HSIL), as compared to Low-grade Intraepithelial Lesion (LSIL).Thus, it may be used in clinical setting to monitor cervical lesions and to predict their progression. Impact statement What is already known on this subject? p16 overexpression is a surrogate biomarker of HPV infection and useful in evaluating HPV-associated squamous and glandular neoplasia of the lower gynaecologic tract. Increased galectin‐3 expression is seen in SCC cervical, with less consistent results in CIN. What do the results of this study add? The results of our study adds to the growing literature that p16 and galectin-3 expression have direct statistically significant correlation with a degree of dysplasia and SCC cervix. Expression of p16 and galectin-3 was more pronounced in invasive SCC and high-grade intraepithelial lesion (HSIL), as compared to low-grade intraepithelial lesion (LSIL). What are the implications of these findings for clinical practice and/or further research? This correction of p16 and galectin-3 expression with degree of dysplasia and SCC cervix can be used for screening and early detection of cervical lesions and thus aid their early treatment and increased survival.

Understanding Anterior Parametrectomy for Preventing Local and Pelvic Recurrences in Cervical Cancer - an Answer to Failure of LACC Trial

Study Objective This video is a demonstration of the anatomy of parametrium and its significance in radical hysterectomy in cervical cancer. Design Video Demonstration. Setting It includes a case of ca cervix stage IB undergoing nerve sparing radical hysterectomy with complete parametrectomy. The dissection of posterior and lateral parametrium is technically easier as compared to that of anterior parametrium. Anteriorly and posteriorly, protection to the spread of cervical cancer is provided by the cervico-vesical fascia and Denonvilliers fascia respectively. Deficiency or the weakest area facilitating tumor spread lies on the lateral aspects as the cervico- vesical ligaments and their incomplete removal leads to local and pelvic recurrences. Patients or Participants 60-year-old with post-menopausal bleeding & cervical biopsy squamous cell carcinoma. FIGO stage IB2. Interventions Understanding the anatomy of parametrium is an important step to achieve optimum tumor clearance. “Lymphatics follow the veins”, was the dictum that led to the origin for the need of parametrectomy in this radical procedure. Schematic diagrams depict the various extents of the parametrium in the three-dimensional scenario. The inferior vesical vein is ligated around two centimeters lateral to the lateral border of vagina making it the lateral extent on either side enforcing complete parametrial clearance. Meticulous and careful dissection of the ureter, achieving hemostasis and preserving the innervation were the various challenges encountered during the procedure. Laparoscopy with its magnified vision and a thorough knowledge of anatomy can achieve a complete parametrectomy. Measurements and Main Results Mean operative time 80 minutes. Blood loss 100ml. Conclusion Thus, this video makes an effort in understanding the anatomy of anterior parametrium. The failure to recognize it or incompetence to achieve it leads to higher rates of recurrences and lower survival rates in minimally invasive approach for cervical cancer as highlighted in the LACC trial.

Abstract 2012: Recurrent rearrangements of NAALADL2 in prostate, breast, cervical, head and neck and lung squamous cell carcinoma

Abstract Prostate cancer is a heterogeneous disease with unique molecular aberrations present in patient sub-groups. Distinct prostate cancer molecular changes have been shown to associate with specific clinical outcomes, suggesting the potential of molecular markers as diagnostic and prognostic biomarkers for prostate cancer. Recurrent ETS family gene fusions, BRAF and SPINK1 overexpression account for about 50-60% of the prostate cancer cases. Genetic aberrations in the remaining 40-50% of the cases is not known. Our attempt to identify new molecular markers in prostate cancer led to the identification of NAALADL2 gene shown to be associated with aggressive prostate cancer. In this study, we carried out a comprehensive analysis of genomic changes of NAALADL2 in prostate and other solid cancers by analyzing copy number changes and gene expression using TCGA next generation RNA sequencing data. We observed recurrent amplification, rearrangement, deletion, mutations and over-expression of NAALADL2 in prostate cancer cases. Notably, we observed mutually exclusive aberrations in NAALADL2 when compared to other cases with known prostate cancer aberrations including ETS gene fusions, indicating NAALADL2 as a distinct molecular sub-set of prostate cancer. Independent validation by fluorescent in situ hybridization (FISH) analysis using break apart probe for NAALADL2 on 874 prostate cancer revealed recurrent amplification and rearrangements in about 8% (71/874) [PN1] of the cases with higher prevalence in Caucasian American than African American cases. Based on these results, we explored prostate cancer TGCA gene fusion database and identified additional cases with gene fusions involving NAALADL2. We selected one of the gene fusions identified in the TCGA database involving NAALADL2-PIK3CA and conducted in vitro functional characterization studies and showed its oncogenic properties. Gene expression microarray analysis of RWPE1 cells transfected with NAALADL2-PIK3CA showed dysregulation of genes involved in cancer related pathways, further suggesting a role for NAALADL2-PIK3CA in prostate cancer development. Based on these studies, we explored the incidence of NAALADL2 gene fusion in other solid cancers including lung squamous cell cancer (LUSC), ovarian cancer, head and neck cancer, cervical cancer and breast cancer shown to have recurrent gene fusions identified in the TCGA gene fusion database. Notably, recurrent rearrangements and amplification are seen in a large subset (45%) of LUSC patients, but not in lung adenocarcinoma, suggesting that NAALADL2 could be developed as a novel biomarker in LUSC. Further validation studies using FISH in our independent cohort of LUSC and breast cancer patients including 79 patients from Ghana confirmed recurrent rearrangements and amplification in a subset of cases. In conclusion, similar to ERG, BRAF and FGFR genes, we show recurrent gene fusions of NAALADL2 across multiple solid cancer with potential applications as a pan cancer molecular marker for cancer diagnosis and a potential target for drug development. Citation Format: Pavithra D. Arachchige, Shannon Carskadon, James Hu, Justin Fernando, Darshan S. Chandrashekar, Nilesh S. Gupta, Sean R. Williamson, Dhananjay A. Chitale, Craig G. Rogers, James O. Peabody, Mani Menon, Tarek A. Bismar, Evelyn Jiagge, Sooryanarayana Varambally, Nallasivam Palanisamy. Recurrent rearrangements of NAALADL2 in prostate, breast, cervical, head and neck and lung squamous cell carcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2012.

Clinical significance of the distance between the cricoid cartilage and upper edge of the tumor using PET-CT in cervical esophageal cancer.

Cervical esophageal squamous cell carcinoma (CESCC) is less common compared with thoracic esophageal cancer, and few studies have investigated the clinicopathological features of CESCC. The present study analyzed 69 patients with CESCC who underwent various therapies at the University Hospital of Kyoto Prefectural University of Medicine between January 2000 and December 2016. The distance between the inferior border of the cricoid cartilage and upper edge of the tumor was evaluated using positron emission tomography and computed tomography. Positive and negative values indicated oral and anal directions, respectively. Using receiver operating characteristic curves, the cut-off value for laryngeal preservation was calculated as −5 mm. According to this value, the patients were divided into two groups: The short group (distance from the cricoid cartilage ≥-5 mm) and long group (distance from the cricoid cartilage <-5 mm). There were no significant differences in clinicopathological factors between the two groups except for body mass index. In univariate analysis, the 3-year overall survival rate was significantly lower in short group (45.4 vs. 79.6%; P=0.009). In multivariate analysis, short group was an independent prognostic risk factor (hazard ratio=2.65; P=0.039). This may be due to lymphatic flow around the cervical esophagus.

Multiplex HPV RNA in situ hybridization/p16 immunohistochemistry: a novel approach to detect papillomavirus in HPV-related cancers. A novel multiplex ISH/IHC assay to detect HPV

BackgroundHigh-risk human papillomavirus (HR-HPV) is notoriously associated with tumor progression in a broad spectrum of malignancies. Detection of HR-HPV is clinically important in the management of HPV-related carcinomas, particularly in cervical cancer and oropharyngeal squamous cell carcinoma (OPSCC). Several methods for HPV detection are currently available including Polymerase chain reaction (PCR)-based techniques, DNA in situ hybridization (ISH), RNA ISH, and p16 immunohistochemistry (IHC). Currently, the guidelines for HPV detection in cervical carcinoma are available, while no clear consensus has not yet been reached on the gold standard for HPV testing in OPSCC. Multimodality testing could help to reliably identify patients with transcriptionally active high-risk HPV-positive.MethodsWe propose a multiplex approach carrying out HPV RNA ISH and p16 IHC on the same slide to detect simultaneously HPV E6/E7 transcripts and p16INK4a overexpression. We tested this assay in two different series one of the cervical cancers with p16-positive, as control, and the other of oropharyngeal squamous cell carcinomas with blind p16 status.ResultsThe multiplex HPV RNA ISH /p16 IHC results in the series both of the cervical cancers and the oral-oropharyngeal cancers were fully concordant with the previous results achieved through the classic p16 IHC and HPV RNA scope carried out on two different slides.ConclusionsOur results suggesting several advantages of this technical approach, namely an easy interpretation fully in the light field, the feasibility in formalin-fixed paraffin-embedded tissue sections, complete automation and a potential wide spreadable for routine testing in several clinical laboratories.

High Risk Human Papilloma Virus Genotype Distribution in Cervical Intraepithelial and Invasive Carcinoma

Background: High-risk (HR) Human Papillomavirus (HPV) is an established cause of cervical cancer. HPV genotype detection is significant in preventing cervical cancers through targeted vaccination our study aimed to identify HRHPV16/18 and non 16/18 in cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma (SCC). Methods: A retrospective study was performed at Pathology Department, BMSI, JPMC, Karachi. About 96 cases of CIN and SCC were included. Analysis of HPV genotypes was performed by DNA extraction, PCR amplification and flow-through hybridization technique. The probes used had a cluster of 13 HRHPV into a group of 3 as HPV HR 1, 2 and 3. Chi square/ Fischer Exact test were applied to observe the association of morphological types of the lesion and expression of HPV genotypes. Results: HPV DNA positivity was 44% in our series. HPV HR 1 was observed in majority of cases (61.9%), followed by HPV 16 in 23.8%, HPV HR 3 in 9.5%, and HPV HR 2 in 4.7% cases respectively. The unique finding was absence of HPV 18 in the series. High grade lesions and invasive cancers showed positivity for HPV HR 1 and HPV 16, while low grade lesions were positive for HPV HR 1, 2 and 3 respectively. Conclusion: HPV HR1 are major causative agents for low and high grade intraepithelial and invasive SCC, followed by HPV 16. Absence of HPV 18 was the novel finding. Our results differ from studies within and outside the region, suggestive of diversified genetic makeup and impact of detection techniques on results.

Cancer-Selective Treatment of Cancerous and Non-Cancerous Human Cervical Cell Models by a Non-Thermally Operated Electrosurgical Argon Plasma Device.

Cold atmospheric plasma (CAP) treatment is developing as a promising option for local anti-neoplastic treatment of dysplastic lesions and early intraepithelial cancer. Currently, high-frequency electrosurgical argon plasma sources are available and well established for clinical use. In this study, we investigated the effects of treatment with a non-thermally operated electrosurgical argon plasma source, a Martin Argon Plasma Beamer System (MABS), on cell proliferation and metabolism of a tissue panel of human cervical cancer cell lines as well as on non-cancerous primary cells of the cervix uteri. Similar to conventional CAP sources, we were able to show that MABS was capable of causing antiproliferative and cytotoxic effects on cervical squamous cell and adenocarcinoma as well as on non-neoplastic cervical tissue cells due to the generation of reactive species. Notably, neoplastic cells were more sensitive to the MABS treatment, suggesting a promising new and non-invasive application for in vivo treatment of precancerous and cancerous cervical lesions with non-thermally operated electrosurgical argon plasma sources.