Integrating Cutting-Edge Methods to Oral Cancer Screening, Analysis, and Prognosis.

Oral cancer (OC) has become a significant barrier to health worldwide due to its high morbidity and mortality rates. OC is among the most prevalent types of cancer that affect the head and neck region, and the overall survival rate at 5 years is still around 50%. Moreover, it is a multifactorial malignancy instigated by genetic and epigenetic variabilities, and molecular heterogeneity makes it a complex malignancy. Oral potentially malignant disorders (OPMDs) are often the first warning signs of OC, although it is challenging to predict which cases will develop into malignancies. Visual oral examination and histological examination are still the standard initial steps in diagnosing oral lesions; however, these approaches have limitations that might lead to late diagnosis of OC or missed diagnosis of OPMDs in high-risk individuals. The objective of this review is to present a comprehensive overview of the currently used novel techniques viz., liquid biopsy, next-generation sequencing (NGS), microarray, nanotechnology, lab-on-a-chip (LOC) or microfluidics, and artificial intelligence (AI) for the clinical diagnostics and management of this malignancy. The potential of these novel techniques in expanding OC diagnostics and clinical management is also reviewed.

Human Papilloma Virus-Associated Oral Pharyngeal Squamous Cell Carcinoma: Prevalence, Prevention, and Awareness of Vaccination in the Indian Population.

Human papilloma virus (HPV), one of the most common sexually transmitted infections, plays a pivotal role in head and neck cancer, primarily oral and oropharyngeal squamous cell carcinomas. HPV is a vaccine-preventable disease that also contributes to cervical cancer. Although HPV vaccination effectively protects the individual against all HPV-associated human carcinomas, the awareness of HPV vaccination and its acceptance is poor in developing nations like India. India has a very high burden of oral cancer, and, unfortunately, the morbidity and mortality rates are also high as the cancer is often detected at an advanced stage. In this review, we explore the prevalence of HPV-associated head and neck squamous cell carcinoma among the Indian population and the awareness of HPV vaccination among Indian youth. Since the prognosis for HPV-associated head and neck squamous cell carcinoma is good, early diagnosis of the cancer is crucial in improving the outcome of the treatment modalities. Efforts are needed to create and increase awareness of HPV vaccination. Routine screening for HPV infection in oral mucosa can prevent the silent epidemic from taking the lives of many young people.

Melatonin: Emerging Player in the Management of Oral Cancer.

Oral cancer (OC) has emerged as a major medical and social issue in many industrialized nations due to the high death rate. It is becoming increasingly common in people under the age of 45, although the underlying causes and mechanisms of this increase remain unclear. Melatonin, as a pleiotropic hormone, plays a pivotal role in a wide variety of cellular and physiological functions. Mounting evidence supports melatonin's ability to modify/influence oral carcinogenesis, help in the reduction of the incidence of OC, and increase chemo- and radiosensitivity. Despite its potential anti-carcinogenic effects, the precise function of melatonin in the management of OC is not well understood. This review summarizes the current knowledge regarding melatonin function in anti-carcinogenesis mechanisms for OC. In addition, clinical assessment and the potential therapeutic utility of melatonin in OC are discussed. This review will provide a basis for researchers to create new melatonin-based personalized medicines for treating and preventing OC.

Association of Interactions between Metabolic 'Caretaker' Genes, p53, MDM2, and Tobacco Use with the Risk of Oral Cancer: A Multifactor Dimensionality Reduction Approach.

BACKGROUND: The present study investigated the association of interactions between gene polymorphisms in metabolic 'caretaker' genes (Phase I: CYP1A1, CYP2E1; Phase II: GSTM1, GSTT1), the cell cycle regulatory gene, p53, along with its negative controller, MDM-2, and the environment variable (tobacco). A nonparametric model, multifactor dimensionality reduction (MDR), was applied to analyse these interactions. MATERIALS AND METHODS: This case-control study was carried out on 242 subjects. Genomic DNA was extracted from peripheral blood lymphocytes.11 gene variants with an exposure variable (tobacco use) were analysed using MDR to identify the best locus model for gene-gene and gene-environment interactions. Statistical significance was evaluated using a 1000-fold permutation test using MDR permutation testing software (version 1.0 beta 2). The value of p<0.05 was considered statistically significant. RESULTS: The best three-locus model for gene-gene interaction included two of the p53 gene polymorphisms; rs17878362 (intron 3) and rs1042522 (exon 4) and rs6413432 in the Phase I gene, CYP2E1(DraI). The three-locus model to evaluate the gene-environment interaction included two intronic polymorphisms of the p53 gene, that is, rs17878362 (intron 3) and rs1625895 (intron 6), and rs4646903 in the Phase I gene CYP1A1*2C. The interaction graphs revealed independent main effects of the tobacco and p53 polymorphism, rs1042522 (exon 4), and a significant additive interaction effect between rs17878362 (intron 3) and rs1042522 (exon 4). CONCLUSIONS: The nonparametric approach highlighted the potential role of tobacco use and variations in the p53 gene as significant contributors to oral cancer risk. The findings of the present study will help implement preventive strategies in both tobacco use and screening using a molecular pathology approach.

Potential of Cytochrome P450, a Family of Xenobiotic Metabolizing Enzymes, in Cancer Therapy.

Significance: Targeted cancer therapy with minimal off-target consequences has shown promise for some cancer types. Although cytochrome P450 (CYP) consists of 18 families, CYP1-4 families play key role in metabolizing xenobiotics and cancer drugs. This eventually affects the process of carcinogenesis, treatment outcomes, and cancer drug resistance. Differential overexpression of CYPs in transformed cells, together with phenotypic alterations in tumors, presents a potential for therapeutic intervention. Recent Advances: Recent advances in molecular tools and information technology have helped utilize CYPs as cancer targets. The precise expression in various tumors, X-ray crystal structures, improved understanding of the structure-activity relationship, and new approaches in the development of prodrugs have supported the ongoing efforts to develop CYP-based drugs with a better therapeutic index. Critical Issues: Narrow therapeutic index, off-target effects, drug resistance, and tumor heterogeneity limit the benefits of CYP-based conventional cancer therapies. In this review, we address the CYP1-4 families as druggable targets in cancer. An emphasis is given to the CYP expression, function, and the possible mechanisms that drive expression and activity in normal and transformed tissues. The strategies that inhibit or activate CYPs for therapeutic benefits are also discussed. Future Directions: Efforts are needed to develop more selective tools that will help comprehend molecular and metabolic alterations in tumor tissues with biological end-points in relation to CYPs. This will eventually translate to developing more specific CYP inhibitors/inducers. Antioxid. Redox Signal. 38, 853-876.

Alterations in p53 Influence hTERT, VEGF and MMPs Expression in Oral Cancer Patients.

BACKGROUND: Mutant p53 is the crucial molecule in the etiopathogenesis of oral cancer. Therefore, we aimed to evaluate the impact of alterations of the p53 gene and its negative feedback regulator, MDM2, on the expression of hTERT, VEGF, and MMPs; the critical genes involved in oral cancer progression. MATERIAL AND METHODS: p53 and MDM2 genotyping were done by PCR-RFLP. p53 mutation analysis was performed using PCR-SSCP and sequencing. hTERT, VEGFA isoforms, MMP2, and MMP9 mRNA levels were analyzed by semi-quantitative Reverse Transcriptase PCR. RESULTS: Arg allele at p53 exon 4 was significantly associated with overexpression of hTERT, MMP2, and MMP9 individually. Expression of hTERT, VEGF A isoforms, MMP2 and MMP9 were significantly altered in the presence of p53 and MDM2 polymorphisms and p53 mutations in a specific combination. Mutant p53, Arg allele at p53 exon 4 locus, and G/G/or T/T genotype at MDM2revealed increased expression of hTERT, VEGF A isoforms, and MMP2/9. CONCLUSION: This study provides evidence that apart from mutant p53, naturally occurring sequence variants in p53codon 72 (Arg72Pro) (rs1042522) and MDM2 (rs2279744) significantly alter the expression of hTERT, VEGF-A isoforms, and MMP2/9 in a specific combination. The differential interaction of codon 72 variants with MDM2, hTERT, VEGF-A isoforms and MMP2/9 play a role in the aggressiveness of oral cancer. The results have important implications for oral cancer progression and should be explored for innovative treatment options.

CRISPR/Cas9: A Molecular Tool for Ovarian Cancer Management beyond Gene Editing.

Ovarian cancer manifests with early metastases and has an adverse outcome, impacting the health of women globally. Currently, this malignancy is often treated with cytoreductive surgery and platinum-based chemotherapy. This treatment option has a limited success rate due to tumor recurrence and chemoresistance. Consequently, the fundamental objective of ovarian cancer treatment is the development of novel treatment approaches. As a new robust tool, the CRISPR/Cas9 gene-editing system has shown immense promise in elucidating the molecular basis of all the facets of ovarian cancer. Due to the precise gene editing capabilities of CRISPR-Cas9, researchers have been able to conduct a more comprehensive investigation of the genesis of ovarian cancer. This gained knowledge can be translated into the development of novel diagnostic approaches and newer therapeutic targets for this dreadful malignancy. There is encouraging preclinical evidence that suggests that CRISPR/Cas9 is a powerful versatile tool for selectively targeting cancer cells and inhibiting tumor growth, establishing new signaling pathways involved in carcinogenesis, and verifying biomolecules as druggable targets. In this review, we analyzed the current research and progress made using CRISPR/Cas9-based engineering strategies in the diagnosis and treatment, as well as the challenges in bringing this method to clinics. This comprehensive analysis will lay the basis for subsequent research in the future for the treatment of ovarian cancer.

Role of the Tumor Microenvironment and the Influence of Epigenetics on the Tumor Microenvironment in Oral Carcinogenesis: Potential Implications.

Oral cancer has become a significant problem throughout the world, particularly in countries that are still developing. Recent literature supports the contribution of components of the tumor microenvironment (TME) and the effect of epigenetic changes happening in the cells of the TME on oral cancer development and progression. In this review, we comprehensively examine the significance of TME in the development of OC along with the current understanding of the epigenetic modifications that regulate the TME and their cohesive impact on tumor traits and their potential as therapeutic targets.

Achieving coronal plane alignment in total knee arthroplasty through modified preoperative planning based on long-leg radiographs: a prospective study.

PURPOSE: This prospective study was undertaken to examine whether the desired coronal plane alignment of limb and prosthetic components in total knee arthroplasty (TKA) could be achieved precisely using conventional jig-based methods by modifying the preoperative planning of bone resection utilizing long-leg radiographs (LLRs). METHODS: The study included consecutive 245 TKA procedures. Pre- and postoperative radiological variables, i.e., the mechanical axis (hip-knee-ankle [HKA] axis), mechanical lateral distal femoral angle (mLDFA), and medial proximal tibial angle (MPTA), and their outliers were evaluated. Statistical analysis was performed using SPSS version 21.0. RESULTS: The mean postoperative HKA axis, mLDFA and MPTA was 179.80 ± 1.81° (p < 0.01; 95% CI: 8.09-9.67), 90.35 ± 1.54° (p < 0.01; 95% CI: 1.33-2.02), and 90.26 ± 1.25° (p < 0.01; 95% CI: 4.41-5.20), respectively. The postoperative HKA axis on the coronal plane was 180 ± 3° in 235 knees (95.92%, 4.08% outliers). Femoral and tibial components were implanted in an acceptable position, withing 90 ± 3° of the mechanical axis of the femur and tibia on the coronal plane in 238 (97.14%, 2.86% outliers) and 243 (99.18%, 0.8% outliers) knees, respectively. CONCLUSION: Modified preoperative planning for TKA on LLRs is a reliable and consistent method to achieve the desired limb and component alignment on the coronal plane without adding financial or logistical costs. LEVEL OF EVIDENCE: II.

"p53 mutation spectrum and its role in prognosis of oral cancer patients: A study from Gujarat, West India".

BACKGROUND AND AIM: p53 mutations are critical players in etiopathogenesis of oral cancer. Interestingly, they show differences in terms of type and codon specificity. These differences might be attributed to geographical variations in tobacco use. We aimed to analyze the frequency of p53 mutations in oral cancer patients from Gujarat, India and their effect on clinico-pathological features, local recurrence and survival. MATERIAL AND METHODS: p53 mutation analysis was performed on 46 paired tissue samples (adjacent normal and primary malignant) using PCR-SSCP and sequencing. RESULTS: Sequencing confirmed 51 p53 mutations in 46 paired tissues. Three novel mutations (frameshift deletion in exon 4; G>T transversion at codon 117 in exon 4 and G>A transition at codon 319 in exon 9) were identified. Distinct pattern of p53 mutations was observed: more common C>T transitions and recurring mutation sites at codon 90 and 116 in exon 4. Interestingly, the probability of developing recurrence was higher in small tumors (<4 cm) with p53 mutations and in cases with p53 mutations in both adjacent normal and malignant tissues. A significant low disease free survival and overall survival was observed in cases harboring truncating and transcriptionally non-active mutations. CONCLUSION: We report a very high frequency and a diverse pattern of p53 mutations in cases from this region. Interestingly, three distinct novel mutations in exons 4 and 9 were also observed. Analyzing p53 mutation status in tumor tissues at an early stage could serve as an important prognostic factor.

Combined evaluation of matrix metalloproteinases and their inhibitors has better clinical utility in oral cancer.

BACKGROUND: Oral cancer is a major health hazard worldwide with increasing incidence and mortality. Cervical lymph node metastasis is a major determinant of outcome in oral cancer. The matrix metalloproteinase (MMP) system is critically involved in invasion and metastasis. Assessment of MMPs and tissue inhibitors of MMPs (TIMPs) in certain combinations might have better clinical efficacy given their potential role in the metastatic process. AIM: Plasma concentrations of MMP-2, MMP-9, TIMP-1 and TIMP-2 in 50 controls and 75 oral cancer patients (nonmetastatic, n=54; metastatic, n=21) were evaluated to assess their investigative value and role in predicting the behavior of this malignancy. METHODS: The plasma concentrations of MMP-2, MMP-9, TIMP-1 and TIMP-2 were quantified by ELISA. The best 2- and 3-marker combinations were calculated using the statistical software mROC. The diagnostic values for all the biomolecules as single markers and their combinations were estimated using the measures of diagnostic accuracy, i.e. the area under the ROC curve and the sensitivity and specificity at cutoff limits with the highest diagnostic accuracy and at the 95% limits of sensitivity and specificity, respectively. RESULTS: MMP-9, TIMP-1 and TIMP-2 were significantly elevated (p=0.000, p=0.013 and p=0.005, respectively) in oral cancer patients. MMP-9 emerged as the best single statistically significant marker in plasma for oral cancer detection. It showed an increase in diagnostic performance when tested in combination with MMP-2 and TIMP-2. The median plasma MMP-9 levels were elevated in both the metastatic and nonmetastatic groups compared with controls (p<0.004 and p<0.007, respectively). CONCLUSION: The results indicated that plasma MMP and TIMP levels in relevant combinations may facilitate clinical decisionmaking for improved management of oral cancer patients and may provide important data for selecting patients for treatment with drugs that interfere with MMP and TIMP activities.

Matrix metalloproteinases and their inhibitors: correlation with invasion and metastasis in oral cancer.

Matrix metalloproteinases (MMPs) have been implicated in invasion and metastasis of various malignancies. The study evaluated a comprehensive profile of MMP-2 and MMP-9 and their inhibitors, tissue inhibitor of metalloproteinases-2 (TIMP-2) and tissue inhibitor of metalloproteinases-1 (TIMP-1), respectively in 50 controls and 75 patients with oral squamous cell carcinoma (OSCC). Blood samples from controls and patients as well as malignant and adjacent normal tissues from the patients were collected. The study examined pro, active and total forms of MMP-2 and MMP-9 using zymography. Enzyme-linked immunoassay (ELISA) and reverse transcription polymerase chain reaction were carried out to evaluate protein levels and mRNA expression; respectively, for the MMPs and TIMPs. Plasma pro, active and total MMP-2, MMP-9 as well as TIMP-1 and TIMP-2 levels were significantly higher in oral cancer patients as compared to the controls. mRNA expression of the MMPs and TIMPs was significantly higher in malignant tissues as compared to adjacent normal tissues. A significant positive correlation was observed between levels of proMMP-9 and active MMP-9 with differentiation, stage and infiltration. ProMMP-2 and active MMP-2 exhibited significant positive correlation with differentiation and lymph node involvement. The multivariate analysis of ELISA results revealed a significant positive correlation between MMP-2, TIMP-1 and TIMP-2 levels with lymph node involvement, stage and differentiation. The receiver operating characteristic curve (ROC) analysis showed that the levels of MMPs and TIMPs have significant discriminatory efficacy to differentiate between controls and patients. The results indicate that MMP-2, MMP-9, TIMP-1 and TIMP-2 have significant clinical usefulness for oral cancer patients. Zymographic analysis is a simple, cost effective, rapid and sensitive alternative assay.