Expression Of Human Papillomavirus Research Articles

Identification of proteins' expression pathway and the effective miRNAs for the treatment of human papillomavirus-induced cervical cancer: in-silico analyses-experimental research.

Cervical cancer is the fourth most common cancer in women. The risk factors for cervical cancer include human papillomavirus (HPV) infection, age, smoking, number of pregnancies, use of oral contraceptives, and diet. However, long-term HPV infection appears to be the main risk factor for developing cervical cancer. This in-silico analysis aims to identify the expression network of proteins and the miRNAs that play a role in the development of HPV-induced cervical cancer. The critical proteins and miRNAs were extracted using the DisGeNET and miRBase databases. String and Gephi were applied to the network analysis. The GTEx web tool was utilized to Identify tissue expression levels. The Enrichr website was used to explore the molecular function and pathways of found genes. Ten proteins, TP53, MYC, AKT1, TNF, IL6, EGFR, STAT3, CTNNB1, ESR1, and JUN, were identified as the most critical shared gene network among cervical cancer and HPV. Seven miRNAs were found, including hsa-mir-146a, hsa-mir-27, hsa-mir-203, hsa-mir-126, hsa-mir-145, hsa-mir-944, and hsa-mir-93, which have a common expression in cervical cancer and HPV. Overall, the gene network, including TP53, MYC, AKT1, TNF, IL6, EGFR, STAT3, CTNNB1, ESR1, and JUN, and Also, hsa-mir-145, hsa-mir-93, hsa-mir-203, and hsa-mir-126 can be regarded as a gene expression pathway in HPV-induced cervical cancer.

High rate of high-risk human papillomavirus among benign and breast cancer patients in Ethiopia.

There have been numerous studies that showed the presence of human papillomavirus (HPV) in breast cancer; nonetheless, there is ongoing debate regarding their association. Given few studies in Ethiopia, we aimed to investigate the magnitude of HPV infection in Ethiopian breast cancer patients. A total of 120 formalin-fixed paraffin-embedded (FFPE) tissue blocks were obtained, and basic demographic, clinical, and histological data were collected from medical records. DNA was extracted from archived FFPE breast tissue specimens using GeneRead DNA FFPE Kit. The AnyplexTM II HPV28 Detection Kit (Seegene, Korea) was used to detect HPV by following the manufacturer's instructions. The SPSS Version 25 was used to enter and analyze data. Among the 120 study participants; HPV (both high-risk and low-risk) was detected in 20.6% of breast cancer and 29.6% of non-malignant breast tumors. The most common genotype was the high-risk HPV 16 genotype. The frequency of HPV was nearly 10-fold higher in estrogen receptor-positive than ER-negative breast cancer. The percentage of HPV in the luminal (luminal A and luminal B) breast cancer subtypes was also much higher than in the non-luminal subtypes (HER-2 enriched and triple-negative breast cancer). This study did not find a significant difference in HPV expression between breast cancer and non-malignant breast tumors; however, the higher percentage of HPV in ER-positive compared to ER-negative breast cancer warrants further attention.

Characterization of a Diverse Set of Conditionally Reprogrammed Head and Neck Cancer Cell Cultures.

To establish and characterize a diverse library of head and neck squamous cell cancer (HNSCC) cultures using conditional reprogramming (CR). Patients enrolled on an IRB-approved protocol to generate tumor cell cultures using CR methods. Tumor and blood samples were collected and clinical information was recorded. Successful CR cultures were validated against banked reference tumors with short tandem repeat genotyping. Cell morphology was archived with photodocumentation. Clinical and demographic factors were evaluated for associations with successful establishment of CR culture. Human papilloma virus (HPV) genotyping, clonogenic survival, MTT assays, spheroid growth, and whole exome sequencing were carried out in selected cultures. Forty four patients were enrolled, with 31 (70%) successful CR cultures, 32% derived from patients who identified as Black and 61% as Hispanic. All major head and neck disease sites were represented, including 15 (48%) oral cavity and 8 (26%) p16-positive oropharynx cancers. Hispanic ethnicity and first primary tumors (vs. second primary or recurrent tumors) were significantly associated with successful CR culture. HPV expression was conserved in CR cultures, including CR-024, which carried a novel HPV-69 serotype. CR cultures were used to test cisplatin responses using MTT assays. Previous work has also demonstrated these models can be used to assess response to radiation and can be engrafted in mouse models. Whole exome sequencing demonstrated that CR cultures preserved tumor mutation burden and driver mutations. CR culture is highly successful in propagating HNSCC cells. This study included a high proportion of patients from underrepresented minority groups. Not Applicable Laryngoscope, 134:2748-2756, 2024.

Single-Cell and Spatial Transcriptomics Revealing the Role of IDO1 in HPV+ Cervical Cancer Tumor Immune Microenvironment and Its Implications in Radiotherapy and Immunotherapy

Persistent infection of human papillomavirus (HPV) is one major etiology of cervical cancer (CC). By now, anti-PD-1 immunotherapy is approved for advanced CC patients, but the response rate was just about 10-20%, tumor immune microenvironment (TIME) might be one factor that affect the efficacy. The indoleamine 2,3-dioxygenase (IDO), a metabolic immune checkpoint, is recently shown to have a correlation-ship with HPV carcinogenesis in CC, with unknown mechanism. This study, using the single cell transcriptomic single-cell sequencing and spatial transcription sequencing analysis/immunologic technology, aimed to exam the role of IDO1 expression in HPV+ CC TIME and explore the changes after radiotherapy. Newly diagnosed advanced HPV- CC and HPV+ CC patients were tested for the tumor and tumor immune microenvironment (TIME) heterogeneity and their changes after fractionated radiation therapy. Tumor tissues were collected, single cell suspension was made for Single-cell RNA sequencing (SCRNAseq) using the 10 × Genomics, while frozen tissue was embedded for spatial transcriptome sequencing (STRNAseq). Seurat 4.0 was used to cluster and annotate cell clusters and map SCRNAseq data to the STRNAseq data. The specific characters of cell clusters were computed by Gene Set Enrichment Analysis (GSEA). SPOTLight and CellChat were used to analyze cell location and interaction respectively. A total of 28631 cells were clustered into 31 cell subsets in HPV- CC and HPV+ CC tissues, including baseline (Pre HPV- CC and Pre HPV+ CC) and 3-week after radiotherapy (Post 3w HPV- CC and Post 3w HPV+CC). There were 10431 epithelial cells (Epi) in all these 4 tumor tissues, with heterogenous IDO1 expression, including IDO1-high Epi, IDO1-low Epi, and IDO1-neg Epi. Interestingly, more than 99% of Epi in Pre HPV- CC tissues were IDO1-neg cells, while more than 99% in Pre HPV+ CC tissue were IDO1-high. Furthermore, the proportion of IDO1-high Epi in Pre HPV+ CC patient dropped to 16.7% after radiotherapy, while the proportion of IDO1-low Epi rase to 63.3%. Using GSEA, the characters of IDO1-high Epi group was shown to have positive regulation of leukocyte chemotaxis and negative regulation of cell adhesion and differentiation. IDO1-high Epi cells also had the hallmark of interferon gamma response. These cells could mainly receive regulative information of interferon gamma pathway from exhausted CD8 T cells, which could affect the apoptosis of tumor cells. This study comprehensively analyzed the immune suppressive role of IDO1-high Epi cells in HPV+ CC TIME at the single-cell transcriptional scale and explored their functional characters in CC radiotherapy. This would be able to provide more evidence to combine with radiotherapy and immunotherapy to improve patients' prognosis.

Identification of HPV oncogene and host cell differentiation associated cellular heterogeneity in cervical cancer via single-cell transcriptomic analysis.

Human Papillomaviruses (HPVs) are associated with around 5%-10% of human cancer, notably nearly 99% of cervical cancer. The mechanisms HPV interacts with stratified epithelium (differentiated layers) during the viral life cycle, and oncogenesis remain unclear. In this study, we used single-cell transcriptome analysis to study viral gene and host cell differentiation-associated heterogeneity of HPV-positive cervical cancer tissue. We examined the HPV16 genes-E1, E6, and E7, and found they expressed differently across nine epithelial clusters. We found that three epithelial clusters had the highest proportion of HPV-positive cells (33.6%, 37.5%, and 32.4%, respectively), while two exhibited the lowest proportions (7.21% and 5.63%, respectively). Notably, the cluster with the most HPV-positive cells deviated significantly from normal epithelial layer markers, exhibiting functional heterogeneity and altered epithelial structuring, indicating that significant molecular heterogeneity existed in cancer tissues and that these cells exhibited unique/different gene signatures compared with normal epithelial cells. These HPV-positive cells, compared to HPV-negative, showed different gene expressions related to the extracellular matrix, cell adhesion, proliferation, and apoptosis. Further, the viral oncogenes E6 and E7 appeared to modify epithelial function via distinct pathways, thus contributing to cervical cancer progression. We investigated the HPV and host transcripts from a novel viewpoint focusing on layer heterogeneity. Our results indicated varied HPV expression across epithelial clusters and epithelial heterogeneity associated with viral oncogenes, contributing biological insights to this critical field of study.

Introduction of E-Score: A gene expression score to characterize immunogenic versus oncogenic HPV expression in HNSCC.

6057 Background: Human Papillomavirus (HPV) integration in the human genome occurs in a majority of HPV(+) head and neck squamous cell carcinomas (HNSCC), with direct and indirect effects on host gene expression and tumorigenesis. Although previous studies showed that HPV integration in HNSCC is overrepresented in genes often somatically mutated in head and neck, lung, and urogenital cancers, reports are mixed for whether patients with HPV integration have worse clinical outcomes. These disparate results may be due to differences in how integration is defined. Integration can often lead to partial loss of the viral genome (e.g. HPV genes E1, E2, or E5) with retention of the oncogenes E6 and E7, which led to a low E2/E7 expression ratio being used as a biomarker for integration. Conversely, others have used the direct detection of DNA or RNA HPV-human breakpoints to identify tumors harboring integrated HPV. HPV integration and the retention or deletion of HPV genes can dramatically change the expression of HPV genes, and we hypothesize it also affects the host immune response. Hypothesis: Loss of non-oncogenic HPV genes relative to E6 and E7 levels reduces immunogenicity, leading to weakened immune cell infiltration and worse clinical outcomes. Methods: Bulk RNA sequencing data from HPV(+) HNSCC tumors (n = 136) was used to characterize the differences in retention or loss of HPV genes and calculate the E-Score as the sum of the logCPM of HPV genes E1, E2, E4, and E5 relative to E7. Cell type deconvolution was performed using ComBat to correct for batch effects, and CIBERSORTx to quantify cell type proportions within the tumor samples. Kaplan-Meier estimates of recurrence and overall survival (OS) were calculated, and Cox proportional hazards tests with covariates were used to test for association with recurrence and OS. To test the correlation between immune cell type inferred proportions and E-Score, T-Tests were performed between samples with high and low E-scores. Results: Low E-Score was significantly associated with recurrence (p = 0.012) but not with OS (p = 0.39) in the multivariate model accounting for age, tumor stage (AJCC8), smoking status, drinking status, BMI, marital status, and molecular tumor subtype (defined as IMU or KRT). For integration positive samples (n = 66), higher E-Score, suggesting lower rates of HPV gene loss, was correlated with higher percentages of dendritic cells (p = 0.044), macrophages (p = 0.021), and CD8+ T Cells (p = 0.024), but not CD4+ T cells (p = 0.14). This significance did not hold when looking at integration negative samples. Conclusions: These results suggest that HPV+ HNSCC patients with a loss of E1, E2, E4 and E5 tend to have less immune cell infiltration and are more likely to have recurrence. We conclude that E-Score is a potential prognostic biomarker of recurrence in HPV+ HNSCC and potential predictive biomarker for immunotherapy.

Cellular states are coupled to genomic and viral heterogeneity in HPV-related oropharyngeal carcinoma.

Head and neck squamous cell carcinoma (HNSCC) includes a subset of cancers driven by human papillomavirus (HPV). Here we use single-cell RNA-seq to profile both HPV-positive and HPV-negative oropharyngeal tumors, uncovering a high level of cellular diversity within and between tumors. First, we detect diverse chromosomal aberrations within individual tumors, suggesting genomic instability and enabling the identification of malignant cells even at pathologically negative margins. Second, we uncover diversity with respect to HNSCC subtypes and other cellular states such as the cell cycle, senescence and epithelial-mesenchymal transitions. Third, we find heterogeneity in viral gene expression within HPV-positive tumors. HPV expression is lost or repressed in a subset of cells, which are associated with a decrease in HPV-associated cell cycle phenotypes, decreased response to treatment, increased invasion and poor prognosis. These findings suggest that HPV expression diversity must be considered during diagnosis and treatment of HPV-positive tumors, with important prognostic ramifications.

Human papilloma virus and tobacco-associated modulation of NOTCH1 pathway during development of head and neck squamous cell carcinoma: Clinical importance

Background: Modulation of NOTCH1 pathway during development of head and neck squamous cell carcinoma (HNSCC) was evaluated with human papillomavirus (HPV) infection and/or tobacco habit to define the molecular mechanism of transformation of normal oral epithelium into neoplastic tissue. Materials and Methods: Comprehensive messenger RNA (mRNA) expression profile of NOTCH1 pathway genes (NOTCH1, JAG1/2, HES1, HEY1, and FBXW7) in HNSCC samples was mined from five different datasets followed by validation at genetic/expression level (RNA/protein). Results: Differential expression pattern of the NOTCH1 pathway genes was seen in different datasets. In our HNSCC samples, high mRNA expression of NOTCH1, JAG1/2, HES1, HEY1 was seen with a significant increase particularly in NOTCH1 and HES1 expression in HPV+ ve (H+) samples. However, no such change in mRNA expression of FBXW7 was noted. In immunohistochemical analysis, high/medium expressions of activated NOTCH1 (NICD)/FBXW7/HES1 proteins were seen in basal/parabasal layers of adjacent normal oral epithelium than in the spinous layers with prevalent expression in H+ samples, followed by increased expression with the progression of the disease. A similar trend was seen in the H+ and/or tobacco+ve (T+) samples, except H-T- group. Infrequent copy number variation was seen in NOTCH1/JAG1/JAG2 loci in HNSCC samples. High/medium expression of NICD/HES1 protein showed overall poor patient outcome, though patients under H+ T– group showed better survival. Conclusion: Our data showed that activated NOTCH1 pathway in basal/parabasal layers of the oral epithelium was gradually aggravated during the development of HNSCC with a distinct association with HPV infection and tobacco habit.

Expression of human papillomavirus 16 and 18 DNA in oral lichen planus using polymerase chain reaction.

Oral lichen planus (OLP) is a chronic inflammatory disease with cell-mediated immune dysregulation. The aetiology of OLP has been studied extensively for decades. Viruses like Hepatitis C Virus (HCV), human papillomavirus (HPV), herpes simplex virus (HSV), and stress have been hypothesized to play a role in the pathogenesis and malignant transformation of OLP. HPV has been proved to be an etiological agent in oropharyngeal cancers and non-tobacco-associated leukoplakia. The role of human papillomavirus in the pathogenesis of OLP has to be studied extensively. This study aims to detect the presence of HPV 16 and HPV 18 DNA in the biopsy samples of OLP and also to determine the role played by the virus in the pathogenesis and malignant transformation of OLP. Biopsy samples comprising 30 OLP tissues were collected. The DNA was extracted by the cetyltrimethylammonium bromide method. Polymerase chain reaction was performed by using general primers to amplify the HPV E6 gene. Twelve out of 30 (40%) OLP cases were positive for HPV DNA. A significant relation was found between HPV, site (buccal mucosa) and the type (reticular) of the lesion (P = 0.007). However, the difference between the percentage of HPV positive males and females was statistically insignificant (P = 0.852). This study confirmed the presence of high-risk HPV 16 and HPV 18 DNA in OLP. The study showed a significantly higher expression of HPV in erosive OLP when compared to reticular OLP, suggesting a possible role of HPV in the malignant transformation of OLP.

Association of Cervical Dysbacteriosis, HPV Oncogene Expression, and Cervical Lesion Progression.

ABSTRACTAs the fourth most common gynecological cancer, cervical cancer has resulted in more than 300,000 deaths worldwide in 2020. The expression of the human papillomavirus (HPV) oncogenes E6 and E7 is significantly involved in the initiation and progression of cervical neoplasia. Additionally, the composition of the vaginal microbiome was also closely associated with the ability of HPV to induce cervical cancer. However, the relationship between the expression of HPV E6/E7 oncogene and the composition of the vaginal microbiome has not been clearly explored. In our present study, to investigate the relationship between the HPV E6/E7 oncogene and vaginal microbiome, cervical swabs from 115 patients were collected, and their vaginal microbiomes were analyzed by using metagenomics sequencing. Along with the progression of cervical lesions, the diversity of cervical flora increased gradually, and the abundance of Lactobacillus decreased. Compared with the HPV group, the prevalence of HPV E6/E7 and oncogene expression level were significantly upregulated in cervical intraepithelial neoplasia (CIN) and cervical cancer (CC) groups. Additionally, a positive correlation between the expression of the HPV oncogene and the genera Sneathia, Salmonella, Leptotrichia, Pseudomonas, and Roseovarius in the HPV group was observed. In the CIN group, the enrichment of the genera Sneathia and Megasphaera was weakly linked with HPV oncogene overexpression. In the CC group, a strong association between the overabundance of the genera Peptostreptococcus and Enterococcus and the high expression of HPV oncogene was also found. The increased diversity of the vaginal microbiota and the decreased Lactobacillus abundance were significantly associated with the severity of cervical disease, and the expression of the HPV oncogene could also be regulated by certain pathogens in different stages of cervical lesions.IMPORTANCE The main findings of this study were that we clarified the associations of the different bacterial species with the expression of human papillomavirus (HPV) oncogenes at different stages of cervical cancer. Along with the severity of cervical lesions, the abundance of the genus and species of Lactobacillus obviously declined, while the aerobic and anaerobic bacteria, as well as the prevalence and expression of HPV E6/E7 oncogene, were increased dramatically.

The Dysregulation of MicroRNAs in the Development of Cervical Pre-Cancer-An Update.

Globally in 2020, an estimated ~600,000 women were diagnosed with and 340,000 women died from cervical cancer. Compared to 2012, the number of cases increased by 7.5% and the number of deaths increased by 17%. MiRNAs are involved in multiple processes in the pathogenesis of cervical cancer. Dysregulation of miRNAs in the pre-stage of cervical cancer is the focus of this review. Here we summarize the dysregulated miRNAs in clinical samples from cervical pre-cancer patients and relate them to the early transformation process owing to human papillomavirus (HPV) infection in the cervical cells. When HPV infects the normal cervical cells, the DNA damage response is initiated with the involvement of HPV’s E1 and E2 proteins. Later, cell proliferation and cell death are affected by the E6 and E7 proteins. We find that the expressions of miRNAs in cervical pre-cancerous tissue revealed by different studies seldom agreed with each other. The discrepancy in sample types, samples’ HPV status, expression measurement, and methods for analysis contributed to the non-aligned results across studies. However, several miRNAs (miR-34a, miR-9, miR-21, miR-145, and miR-375) were found to be dysregulated across multiple studies. In addition, there are hints that the DNA damage response and cell growth response induced by HPV during the early transformation of the cervical cells are related to these miRNAs. Currently, no review articles analyse the relationship between the dysregulated miRNAs in cervical pre-cancerous tissue and their possible roles in the early processes involving HPV’s protein encoded by the early genes and DNA damage response during normal cell transformation. Our review provides insight on spotting miRNAs involved in the early pathogenic processes and pointing out their potential as biomarker targets of cervical pre-cancer.

Association of Human Papilloma Virus in Oral Squamous Cell Carcinoma

Introduction: Oral squamous cell carcinoma is the most common malignant epithelial neoplasm affecting the oral cavity.it is commonly seen in the lip, oral cavity, salivary glands. Worldwide, oral cancer accounts for 2%–4% of all cancer cases. Although tobacco and alcohol are the main etiologic factors in majority of these cancers, the etiological factor in 1/4th of the cases remains unidentified. There is a growing evidence that human papilloma virus (HPV) may act as a cocarcinogen along with tobacco which eventually results in oral cancers.
 Hence, this study is focused on evaluating the expression of HPV in OSCC and to correlate the association of HPV in OSCC patients.
 Aim: The aim of this study is to analyze the presence of human papilloma virus in oral squamous cell carcinoma using p16 tumor biomarker and to compare the presence of human papilloma virus in tobacoo related and non tobacoo related oral squamous cell carcinoma.
 Methodology: A 100 histologically diagnosed OSCC patients were selected for the study and sections were made to evaluate the presence of HPV using P16 tumor marker.
 Results: Out of the total 100 patients the presence of HPV was seen in only 3 patients. Tongue was the common site with a male predilection. Even though non smokers were more in the study, the presence of HPV was seen in patients with smoking habit. Thus correlating the relationship between presence of HPV and smoking habit.
 Conclusion: There is no significant association between OSCC and HPV in our study. Our study also proves that there is an association between HPV and smoking habit. However considering our sample size, a larger sample size can be consider to prove the association between OSCC and HPV.

ERa-36 instead of ERa mediates the stimulatory effects of estrogen on the expression of viral oncogenes HPV E6/E7 and the malignant phenotypes in cervical cancer cells

High risk human papillomavirus (HPV) is the main causative factor of cervical cancer. In addition, estrogen and its receptors are also involved in the development of carcinogenesis. The canonical estrogen receptor ERα is frequently deficient while its variant ERα-36 is highly expressed in cervical cancer cells. The biological significance for this receptor transition from ERα to ERα-36 remains unclear. In the present study, the results of RT-PCR and Western blot demonstrated that ERα and ERα-36 function antagonistically on the expression of the viral oncogenes HPV E6 and E7. At mRNA and protein levels, ERα inhibited HPV E6/E7 expression whereas ERα-36 stimulated HPV E6/E7 expression. Overexpression of ERα-36 promoted cell proliferation while reintroduction of ERα into cervical cancer cells did not significantly affect cell proliferation which is in line with the different effects of . ERα-36 and ERα on the expression of cell cycle regulator, namely p53, p21 and cyclin D1. Furthermore, ERα suppressed whereas ERα-36 promoted the migration and invasion of cervical cancer cells, which should be related to the oppositive roles of ERα and ERα-36 in the Wnt/β-catenin/MRTF-A signaling pathway which is activated by HPV E7. Results of this study suggest that ERα functions as a tumor suppressor whereas ERα-36 is an oncoprotein in cervical cancer cells. ERα deficiency together with ERα-36 overexpression might enhance the expression of HPV E6/E7 genes and facilitate the development of cervical cancer. Targeting ERα-36 with selective antagonists should be a promising strategy for cervical cancer therapy.

Oncogenic E6 and/or E7 proteins drive proliferation and invasion of human papilloma virus‑positive head and neck squamous cell cancer through upregulation of Ror2 expression.

Ror2 (receptor tyrosine kinase like orphan receptor 2) is highly expressed in various types of cancers; in the majority of these cancers, Ror2 expression is associated with more aggressive disease states. Recently, it has been reported that Ror2 is highly expressed in human papilloma virus (HPV)‑positive head and neck squamous cell cancer (HNSCC) cell lines, presumably indicating that Ror2 plays a critical role in HPV‑related cancers. However, the function of Ror2 in HPV‑positive HNSCC is currently unknown. Here, we first examined the expression levels of Ror2 in clinical specimens from patients with HPV‑negative and HPV‑positive oropharyngeal squamous cell cancer (OPSCC) via immunohistochemical analysis. We found that Ror2 was expressed in both HPV‑negative and HPV‑positive OPSCC tissues. We then confirmed that HPV‑positive HNSCC cell line, UPCI:SCC152 cells, express Ror2 higher than HPV‑negative cell lines as previously reported. Suppressed expression of HPV E6/7 resulted in reduced expression levels of Ror2. We also revealed that Ror2 downregulation significantly inhibited the proliferation of UPCI:SCC152 cells without inducing apoptosis. Moreover, Ror2 knockdown decelerated G1/S phase progression and abrogated invasive migration of UPCI:SCC152 cells. These results provide strong evidence that E6 and/or E7 oncoproteins regulate the progression of HPV‑positive HNSCC by upregulating Ror2 expression, suggesting that Ror2 could potentially be a novel target in HPV‑related cancers.

The Expression of E6 HPV, P53 and P16ink4a at Well, Moderately, and Poorly Differentiated Cervical Adenocarcinoma

The objective of this study is to analyze the expression of E6 Humanpapilloma virus (HPV), p53, and p16INK4A in cervical adenocarcinoma grade well differentiated (WD), moderately differentiated (MD), and poorly differentiated (PD). A cross sectional study conducted at Department of Anatomic Pathology, Dr. Soetomo General Academic Hospital Surabaya Indonesia using formalin fix paraffin embedded (FFPE) from cervical normal and cervical adenocarcioma grade WD, MD, and PD. The expression of E6 HPV, p53, and p16INK4A was performed by immunohistochemistry (IHC) staining. Data were analyzed with Kruskal-Wallis and continued with Mann-Withney test. The expression of E6 HPV in the cervical adenocarcinoma showed 35.9% specimens represented negative and 64.1% specimens represented positive. There was no significant difference in the expression of E6 HPV and p53 in cervical adenocarcinoma between grade WD, MD, and PD. The p16INK4A was overexpressed, shown as diffuse appearance in 89.7% of the specimens. There was a significant difference in the expression of p16INK4A between grade WD and MD with PD. In conclusion, some of cervical adecarcinoma were not caused by infection of HPV type 16 or 18 and the expression of p16INK4A might take a role in the developing of malignancy that caused by infection of HPV.

Sonic Hedgehog is a novel prognostic biomarker in patients with oral squamous cell carcinoma

Recent studies revealed that Sonic Hedgehog (SHH) signaling pathway plays an important role in initiation and tumor progression in various malignancies, however, its role in oral squamous cell carcinoma (OSCC) remains unclear. The objective of this study was to investigate the prognostic significance of SHH expression in patients with OSCC in relation to p53 protein expression and HPV (Human Papilloma Virus) presence. SHH, p53, HPV expression was analyzed in surgical specimens from 70 patients with primary OSCC by immunohistochemistry (IHC) and correlated with clinicopathological parameters. The presence of SHH, p53, HPV was found in 51/70 (72.9%), 32/70 (45.7%), 11/70 (15.7%) cases, respectively. No correlation between SHH, p53 overexpression, and HPV presence was found. SHH expression was associated with tumor stage (p = 0.026). P53 immunoreactivity correlated with tumor grade (p = 0.040). By Kaplan-Meier analysis, SHH expression was significantly associated with shorter overall survival (p = 0.005). Multivariate cox regression analysis showed that SHH was an independent prognostic factor for overall survival (HR = 2.93; 95% CI = 1.40-6.13; p = 0.004). Our findings revealed that the SHH signaling pathway contributes to the poor survival of patients with OSCC and should be considered as a new prognostic biomarker in patients with OSCC. Inhibition of the SHH pathway may be used as a new potential target in cancer therapy.

Association of Human Papilloma Virus in Oral Squamous Cell Carcinoma: An Alarming Need for Human Papillomavirus 16 Screening in Cancer Patients.

Background:The percentage of cancers of the tongue and palatine tonsils has continued to increase by 2%–4% among younger men. This increased prevalence of a subsection of oropharyngeal carcinoma can be associated with human papillomavirus (HPV). Among the head-and-neck cancers, a strong association with HPV infection is evident with oropharyngeal cancers, particularly tonsillar and basal tongue cancers.Objectives:Oral carcinoma, with an overall incidence of 16.1 adults per 100,000, is one of the leading malignancies worldwide, presenting a noticeable geographic variation in its distribution. Squamous cell carcinoma (SCC) being the most common of all oral malignancies, the objective of the study is to detect the HPV antigen p16 over-expression in patients with oral SCC using immunohistochemistry (IHC).Materials and Methods:Oral SCC (OSCC) diagnosed formalin-fixed-paraffin embedded blocks were processed for IHC.Results:Out of 50 cases, 3 were deferred due to insufficient tumor sample and 2/47 cases were p16 positive and the site was the lateral border of the tongue.Conclusion:The HPV antigen overexpression in patients with OSCC was investigated to detect the incidence of HPV in SCC of oral cavity. P16 was used as a marker for the detection of OSCC using IHC in HPV-induced OSCC. Positives were detected thus concluding the significance of studying HPV expression during diagnosis.

RPRD1B is a potentially molecular target for diagnosis and prevention of human papillomavirus E6/E7 infection-induced cervical cancer: A case-control study.

The objective of the study is to investigate the biomarkers for diagnosis and prevention of human papillomavirus (HPV) infection-induced cervical cancer. Cervical cancer tissues were collected from patients with cervical cancer, while noncancer tissues were collected from patients diagnosed with cervical lesions or uterine fibroids at the Chinese PLA General Hospital 301 and 309, China from December 2017 to June 2018. The cancer tissues were collected from the site of lesion, while the noncancer tissues were collected from similar anatomical locations. Quantitative real-time PCR, Western blot (WB), and immunohistochemistry (IHC) were used to detect the mRNA and protein levels of HPV E6/E7, RPRD1B (regulation of nuclear pre-mRNA domain containing 1B), cyclin D1, and transcription factor 4 (TCF4) between cervical cancer tissues and noncancer tissues. The correlation of HPV E6/E7, RPRD1B, cyclin D1, and TCF4 expressions was analyzed. Twenty patients with cervical cancer and 27 controls without cervical cancer were included in this study. The mRNA expression of HPV E6/E7and RPRD1B was significantly higher in patients with cervical cancer than controls, while cyclin D1 mRNA expression was significantly lower in patients with cervical carcinoma in situ stage, compared with controls. RPRD1B protein expression was significantly higher in patients compared to controls when analyzed by IHC. TCF4 was significantly lower in clinical stage I and Ib of cervical cancer when analyzed by WB. The mRNA and protein expressions of RPRD1B and cyclin D1 were significantly different between patients younger than 50 years old, compared to patients 50 years and older. HPV E6/E7 expression was associated with RPRD1B level in cervical cancer. The expression of RPRD1B and cyclin D1 in patients with cervical cancer might be affected by age.

PD-L1 Expression and a High Tumor Infiltrate of CD8+ Lymphocytes Predict Outcome in Patients with Oropharyngeal Squamous Cells Carcinoma

Carcinogenesis of human papillomavirus (HPV)-related (+) oropharyngeal squamous cell carcinoma (OPSCC) differs from HPV-negative (–) OPSCC. HPV-related immune-escape-mechanism could be responsible for the development and progression of HPV+ tumors and an immunophenotype different from HPV– OPSCC is expected. The purpose of this study was to analyze the expression of programmed cell death protein 1 ligand 1 (PD-L1) and its prognostic relevance in relation to CD8+ tumor infiltrating lymphocytes (TILs) and the major histocompatibility complex (MHC) I expression in OPSCC. We quantified PD-L1 expression on tumor cells (TC) and macrophages and MHC I expression in association to CD8+ TILs by immunohistochemistry on tissue microarray derived from 171 HPV+/-OPSCC. HPV-status was determined by p16INK4a immunohistochemistry/HPV-DNA detection. Presence of CD8+ TILs, PD-L1 expression on TC, and a more frequent loss of MHC I in HPV+ compared to HPV- OPSCC was detected. A high amount of CD8+ TILs in the whole cohort and in HPV+ OPSCC and PD-L1 expression on TC in HPV- OPSCC was associated with favorable overall survival. There was a trend for an improved outcome according to PD-L1 expression (macrophages) in HPV+ OPSCC without reaching statistical significance. CD8+ TILs and PD-L1-expression have prognostic impact in OPSCC and might present useful biomarkers for predicting clinical outcome and personalized therapy concepts.

Early posttherapy clearance of human papillomavirus and treatment response in cervical carcinoma.

Among patients with cervical cancer, little is known about the significance of persistent human papillomavirus (HPV) expression after chemoradiation (CRT). This study evaluated associations between early posttreatment HPV clearance and patient outcomes with an added focus on the value of posttherapy positron emission tomography (PET) imaging. Included patients underwent pretreatment testing indicating a high-risk HPV infection and posttreatment testing with a messenger RNA (mRNA)-based genital swab after CRT. Posttherapy responses were stratified on the basis of HPV mRNA detection into an early clearance (EC) group (no mRNA) and a persistent expression (PE) group (detectable mRNA) on the basis of an evaluation at a median of 6weeks after therapy. The Kaplan-Meier method was used to compare outcomes, and multivariable analysis was used to identify predictors of outcomes. Seventy-two of the 97 eligible patients (74.2%) had EC. The mean follow-up time was 25months (range, 4-56months), and 2-year pelvic control (76.9% vs 50.2%; P=.01) and overall survival (OS; 80.9% vs 52.2%; P<.01) were superior among EC patients. In the multivariable analysis, EC predicted for improved survival (hazard ratio [HR] for mortality, 0.46; 95% confidence interval [CI], 0.21-0.96; P=.047), as did a complete response (CR) on posttherapy PET (HR for less than a CR on PET, 6.17; 95% CI, 2.58-14.72; P<.01). In a subset analysis of patients with a posttherapy PET CR, HPV clearance retained prognostic significance (2-year OS, 95.6% with EC vs 66.7% with PE; P=.04), whereas PE patients without a PET CR had the worst survival (35.9%; P<.01 for trend). Early posttherapy clearance of HPV is associated with improved survival in cervical cancer. Evaluating HPV expression at this 6-week time point provides prognostic information beyond posttherapy PET imaging and may aid in risk stratification and decisions for treatment escalation.