Salivary MMP-9 as a Biomarker for the Diagnosis of Oral Potentially Malignant Disorders and Oral Squamous Cell Carcinoma

Objective: To compare the salivary MMP – 9 concentration among subjects with oral squamous cell carcinoma (OSCC), oral potentially malignant disorders (OPMD), tobacco users, and control groups. Materials and methods: A total of 88 subjects were enrolled and divided into four study groups viz., OSCC (n=24), OPMD (n=20), tobacco habits (n=22), and healthy controls (n=22). All subjects gave unstimulated saliva samples for the evaluation MMP – 9 by ELISA kit. Demographic information like age, gender, type of tobacco, and duration of the habit were recorded. Results: Subjects with OSCC and OPMD had significantly higher mean MMP-9 levels than subjects with tobacco habits and control groups (P<0.001). Also, poorly differentiated OSCC group had significantly higher mean saliva MMP-9 than moderate and well-differentiated OSCC. The optimal cut-off point was 214.37 ng/mL with a sensitivity of 100% and specificity of 59% for OSCC versus the control group. The optimal cut-off point was as 205.87 ng/mL with a sensitivity of 100% and a specificity of 54% for OPMD versus the control group. Conclusion: The data obtained from this study indicated that OSCC and OPMD had an increased level of salivary MMP-9. Salivary MMP-9 could be a useful, non-invasive adjunct technique in the diagnosis, treatment, and follow-up of OSCC and OPMD.     

Salivary Tumour Necrosis Factor-α as a Biomarker in Oral Leukoplakia and Oral Squamous Cell Carcinoma

Background: Oral cancer is one of the life threatening disease which requires an availability of a biomarker for itsearly detection and also for effective treatment strategies. The current study is done to evaluate the efficacy of one suchbiomarker i.e. TNF- α as an indicator for oral precancer and oral cancer. Objectives: To evaluate the efficacy of Tumournecrosis factor - alpha (TNF)-α as a salivary biomarker in histopathologically diagnosed cases of oral leukoplakia andOral squamous cell carcinoma. To correlate the levels of TNF- α with varying histologic grading in Oral SquamousCell Carcinoma and dysplasia grading in Oral leukoplakia or Hyperkeratosis. Materials and Methods: The studygroup included 90 subjects that were divided into three groups. OSCC (n=30), leukoplakia (n=30) and controls (n=30).Cases were selected based on inclusion and exclusion criteria of the study. Salivary samples were then collected fromall three groups. Salivary levels of TNF-α were estimated using Enzyme Linked Immunosorbent Assay (ELISA). Thedata on concentration gradients obtained were subjected to appropriate statistical analysis. Results: The results of thepresent study demonstrated higher levels of salivary TNF-α in individuals with OSCC compared to leukoplakia andhealthy control subjects with a high level of statistical significance. ROC curve analysis along with diagnostic parametercalculation also revealed that salivary TNF-α to be a better medium for detecting OSCC. There is also an increase inthe salivary TNF-α levels with increase in the histological grade of differentiation in OSCC as well as leukoplakia.Conclusion: The present study concludes that salivary TNF – α can be used as a prognostic biomarker of OSCC. Inview of the elevated levels of TNF – α in saliva of individuals with severe dysplasia, it can also be used to monitor themalignant transformation to leukoplakia to OSCC.     

Diagnostic Accuracy of Salivary Biomarkers in Detecting Early Oral Squamous Cell Carcinoma: A Systematic Review and Meta-Analysis

Background: Biopsy is the gold standard for oral squamous cell carcinoma (OSCC) diagnosis. Salivary biomarkers provide promising complementary alternative diagnostic adjunct for its simple non- invasive collection and technique and to screen large population. Objective: To summarize and compare the existing evidence on diagnostic accuracy of salivary biomarkers with their estimation method in detecting early oral squamous cell carcinoma. Methods: The review protocol is registered under PROSPERO(CRD42021225704). PubMed, Google Scholar, EBSCOhost were searched from 2000 to 2020 to identify the screening potential of eight salivary biomarkers: mRNA, miRNA, DUSP100, s100P, IL-8, IL-1B, TNF-a and MMP-9. True-positive, false-positive, true-negative, false-negative, sensitivity, specificity values were extracted or calculated if not present for each study. Quality of selected studies was evaluated based on QUADAS 2 tool. Meta-analysis was performed using a bivariate model parameter for the sensitivity and specificity and summary points, summary receiver operating curve (SROC), confidence region, and prediction region were calculated. Results: Eighteen studies were included for qualitative synthesis and out of that 13 for meta-analysis. Sensitivity and specificity were calculated with AUC. For mRNA it was 91% and 90% with 0.96 AUC, miRNA had 91% and 91% with 0.95 AUC for PCR. IL-1B had 46% and 60% with 0.61 AUC, S100p had 45% and 90% with 0.57 AUC for ELISA. IL-8 had 54% and 74% for ELISA and 89% and 90% for PCR with 0.79 AUC and DUSP1 had 32% and 87% for ELISA and 76% and 83% for PCR with 0.83 AUC respectively. Conclusion: Early detection of OSCC was best achieved by screening for salivary mRNA and miRNA estimated by PCR. Further investigation is required into salivary RNA as novel biomarkers and these salivary biomarkers may be potentially used for non-invasive diagnosis of early OSCC.     

Protein-Protein Interaction Network Analysis of Salivary Proteomic Data in Oral Cancer Cases

Background: Oral cancer is a frequently encountered neoplasm of the head and neck region, being the eight mostcommon type of human malignancy worldwide. Despite improvement in its control, morbidity and mortality rates haveimproved little in the past decades. Therefore, prevention and/or early detection are a high priority. Proteomics withnetwork analysis have emerged as a powerful tool to identify important proteins associated with cancer developmentand progression that can be potential targets for early diagnosis. In the present study, network- based protein- proteininteractions (PPI) for oral cancer were identified and then analyzed for use as key proteins/potential biomarkers.Material and Methods: Gene expression data in articles which focused on saliva proteomics of oral cancer werecollected and 74 candidate genes or proteins were extracted. Related protein networks of differentially expressed proteinswere explored and visualized using cytoscape software. Further PPI analysis was performed by Molecular ComplexDetection (MCODE) and BiNGO methods. Results: Network analysis of genes/proteins related to oral cancer identifiedkininogen-1, angiotensinogen, annexin A1, IL-8, IgG heavy variable and constant chains, CRP, collagen alpha-1 andfibronectin as 9 hub-bottleneck proteins. In addition, based on clustering with the MCODE tool, vitronectin, collagenalpha-2, IL-8 and integrin alpha-v were established as 5 distinct seed proteins. Conclusion: A hub-bottleneck proteinpanel may offer a potential /candidate biomarker pattern for diagnosis and treatment of oral cancer disease. Furtherinvestigation and validation of these proteins are warranted.     

Oral Cancers in Mumbai,India: a Fifteen Years Perspective with Respect to Incidence Trend and Cumulative Risk

Objective: We estimated the time trends in the incidence and the risk of developing an oral cancer in Mumbai, ‍Indian population using the data collected by the Bombay Population Based Cancer Registry during the 15 year ‍period from 1986 to 2000. ‍Methods: A total of 9,670 oral cancers (8.2% of all neoplasms) were registered, of which 6577 were in males and ‍3093 in females (10.7% and 5.4% of the respective totals for the two genders) . For evaluation of the trend, we ‍applied a linear regression model based on the logarithm of the observed incidence rates. The annual percentage ‍changes were also computed for the incidence rates to evaluate the time trend. ‍Results: In males, a statistically significant decreasing trend in the overall age-adjusted incidence rates were ‍observed during the period 1986 to 2000, with an yearly decrease of 1.70%. This decrease was significant for men ‍above the age of 40, but for young adult men below the age of 40, there was no significant decrease, the level being ‍stable. In females, the overall decreasing trend in the age-adjusted incidence rates of oral cancers was not significant, ‍but in the age group 40-59, a significant decline was observed. The probability estimates indicated that one out of ‍every 57 men and one out of every 95 women will contract any oral cancer at some time in their whole life and 97% ‍of the chance is after he or she completes the age of 40. ‍Conclusion: The observed decreasing trend in oral cancers in Indian men may be attributed to a decrease in the ‍usage of pan and tobacco. The high prevalence of the usage of smokeless tobacco among young adult men and ‍women may explain the stable trend in oral cancer incidence in this group. These findings help to strengthen the ‍association between tobacco use and oral cancer risk. ‍     

Multiple Primary Cancers in Patients with Squamous Cell Carcinoma of the Oral Cavity

During the 27 years between 1962 and 1988,984 patients visited the National Cancer Center Hospital for previously untreated squamous cell carcinoma of the oral cavity and lip. The records of all these 984 patients were reviewed to determine the incidence of additional primary carcinoma. 1) One hundred and thirty-five additional carcinomas developed in 111 patients (11.2%) during 5,689.2 person-years of observation. The incidence of additional primary carcinoma was 23.7 per 1,000 person-years. 2) The cumulative rate of additional primary carcinoma during the first five years of observation showed a tendency to increase in the most recently treated patients (from 1980 to 1988). 3) The observed-to-expected ratio (O/E ratio) for all sites was 2.77 and this is significantly high ( P < 0.01). The calculation of the O/E ratio for each site revealed significantly high risks in the oral cavity and pharynx, esophagus and skin. The O/E ratio for the oral cavity and pharynx was extremely high (79.45). 4) The O/E ratio for all sites in each year of follow-up was the highest in the first year, stayed nearly constant from the second to 14th years, and decreased gradually afterwards. Significantly high risk was observed until the 13th year of follow-up. Patients with oral squamous cell carcinoma must be under frequent and regular examination for almost 15 years.     

Intratumoral Heterogeneity in Breast Cancer: A Comparison of Primary and Metastatic Breast Cancers

Intratumoral heterogeneity presents challenges in the management of cancer. To gain deeper insight in intratumoral heterogeneity at different levels and tumor sites for common biomarkers in breast cancers, this report examines seven cases of invasive breast cancer with multiple axillary nodal metastases and/or recurrences for immunohistochemical expression of estrogen receptors, progesterone receptors, human epidermal growth receptor 2, and Ki67 on all tissue blocks in both primary and metastatic tumors.     

Frequency of Delayed Diagnosis of Oral Squamous Cell Carcinoma in Pakistan

Objectives: To determine the frequency of delayed diagnosis of oral squamous cell carcinoma in our setup; highlighting factors responsible for any delay and their possible relevance to demographic and diagnostic features. Methods: This cross sectional study of six months duration was conducted in the Oral and Maxillofacial Surgery Department of the Armed Forces Institute of Dentistry, Rawalpindi, Pakistan. A total of 246 patients, both male and female, having a biopsy proven definitive diagnosis of OSCC were included using a consecutive sampling technique. Delay in diagnosis was assessed from the stated period of time from when the patient first noticed symptoms of disease until a definitive diagnosis was made. We concluded delayed diagnosis if this was more than 40 days. Results: The ages of patients ranged from 27 to 60 years with a mean of 46.7 ± 10.2 years and a marked male predominance (3.7:1). Delayed diagnosis was observed in 91.5% of cases. However, statistically no significant differences were found with age, gender, marital, education status, household income and time of biopsy. Conclusion: Our primary finding of delayed diagnosis with no prior contact with any health care professional clearly reflects a need of taking urgent measures to avoid serious impacts on morbidity and mortality associated with OSCC.     

Stathmin! An immunohistochemical analysis of the novel marker in Oral Squamous Cell Carcinoma and Oral Leukoplakia.

Background: Stathmin is an intracellular phosphoprotein that controls the microtubule dynamics by further regulating proper attachment and alignment of chromosomes in a dividing cell. Thus, any mutation or aberrantly expressed protein that reduces the fidelity of spindle assembly will enhance chromosomal instability contributing to aneuploidy. Oral Squamous Cell Carcinoma is an extensively studied malignancy that occurs due to accumulated genetic changes due to carcinogens. The current study is done to evaluate the stathmin role and its expression in OSCC and Oral epithelial dysplasia (OED). Objective: The aim of the present study is to evaluate the role of stathmin in OSCC and Oral dysplasia and also to correlate the expression of Stathmin with respect to the different histopathological grades of OED and OSCC. Materials and Methods: 30 neutral buffered formalin fixed, paraffin embedded (FFPE) tissues of Oral Leukoplakia/OED and 30 FFPE tissues of OSCC were subjected to immunohistochemistry with stathmin antibody. Five fields of each case with 300 cells were examined and a mean percentage of positive–stained slides were determined. The percentages were recorded accordingly with their respective histological grades. The results were analysed statistically. Results: The results of the present study demonstrated higher mean values of stathmin in tissues with OSCC (2.50) compared to leukoplakia (2.11) and normal tissues (0.00) with a high level of statistical significance (0.0001). There is also an increase in the percentage levels of stathmin with increase in the histological grade of differentiation in OSCC as well as leukoplakia. Conclusion: The present study found a statistical correlation between increased grades of the disease with expression levels of stathmin. This confirms that stathmin expression can contribute to disease progression and that stathmin might have a potential role as an early diagnostic biomarker and can be a therapeutic target for OSCC.     

Multiple Primary Cancers with Microsatellite Instability: Report of a Case

We report here a patient who developed a variety of tumors both synchronously and metachronously over a 2-year period. The involved organs were the uterus, ureter, and small and large intestines. The patient underwent open surgery 3 times and polypcctomies 6 times. Postoperative histopathologic analysis showed 2 adenomas and 8 carcinomas. Genetic analysis revealed microsatellite instabilities at the tested loci in all 10 tumors, indicating that replication errors played an essential role in the tumorigenesis. Early identification of microsatellite instability could be useful for predicting development of additional primary cancers.     

1393例乳腺癌雌孕激素受体状况分析

目的:探讨乳腺癌患者雌孕激素受体状态分布及与临床指标的关系。方法:采用SP二步法对1393例乳腺癌患者的术后肿瘤石蜡标本进行雌、孕激素受体检测,并与患者的年龄、月经情况、原发肿瘤病理类型和腋淋巴结转移状况相结合进行分析。结果:1)ER和PR的表达呈显著正相关(P<0.0001);2)ER(+)、PR(+)及PR(-)表达率和年龄分布无关(P>0.05),≤39岁年龄组ER(-)表达率明显高于其他年龄组(P<0.001);3)绝经后ER(+)率高于绝经前患者(P<0.0001),PR(+)率低于绝经前患者(P<0.0001);4)组织病理学类型和ER、PR之间关系无显著差异(P>0.05);5)无腋淋巴结转移患者的ER(+)、PR(+)表达率显著高于有转移者(P<0.0001),ER(-)、PR(-)表达率低于有腋淋巴结转移者(P<0.0001)。结论:乳腺癌患者ER、PR的分布有一定的规律,可作为制定综合治疗方案的依据,同时对判断预后有一定意义。     

Epidemiology of Nasopharyngeal Cancers in Iran: A 6-year Report

Background: Nasopharyngeal cancer is a disease with distinct ethnic and geographical distribution. The aimof this review was to describe the epidemiological characteristics of nasopharyngeal cancer in Iran from 2004to 2009 because no systematic study has been performed to evaluate the trends of its incidence yet. Materialsand Methods: The data were derived from the databases of the National Cancer Data System Registry in theperiod of 2004-2009. Nasopharyngeal cancers were classified according to the International Classification ofDiseases for Oncology. Incidence rates and trends were calculated and evaluated by gender, age decade, andhistopathology types. Results: A total of 1,637 nasopharyngeal cancers were registered in Iran from 2004 to 2009giving an incidence of 0.38 per 100,000. The male-to-female ratio was 2.08:1. The trend of incidence was found tohave increased, with a significant increase observed in males. Undifferentiated carcinoma was the most commonhistopathology type in all the age decades. Conclusions: Because the incidence of nasopharyngeal cancers inIran has increased, especially in males, further studies are recommended for understanding of the etiologicalfactors involved in the rise of the disease.     

Candidate Biomarkers for Oral Squamous Cell Carcinoma: Differential Expression of Oxidative Stress-Related Genes

Background: Alteration in the biotransformation of exogenous compounds can result in production of reactiveoxygen species (ROS), which can predispose cells to malignant transformation in the head and neck. This study aimedto evaluate the expression of genes involved in antioxidant metabolism in the oral squamous cell carcinoma (OSCC).Methods: The expression of eighty-four genes was evaluated in OSCC and non-tumor tissues by quantitative real-timepolymerase chain reaction using the TaqMan Gene Expression Array. The biological mechanisms related tothe differentially expressed genes were investigated using Gene – NCBI, KEGG, UNIPROT and REACTOME databases.Results: Twenty-one genes encoding enzymes involved in antioxidant metabolism were differentially expressed inthe OSCC case. Four genes (ATOX1, PRDX4, PRNP, and SOD2) were up-regulated, and seventeen (ALOX12, CAT,CSDE1, DHCR24, DUOX1, DUOX2, EPHX2, GLRX2, GPX3, GSR, GSTZ1, MGST3, PRDX1, OXR1, OXSR1,SOD1, and SOD3) were down-regulated. We identified 14 possible novel biomarkers for OSCC. The differentiallyexpressed genes appeared related to important biological processes involved in carcinogenesis, such as inflammation,angiogenesis, apoptosis, genomic instability, invasion, survival, and cell proliferation. Conclusions: Our studyidentified novel biomarkers which might warrant further investigation regarding OSCC pathogenesis since the alteredexpression in the genes can modulate biological processes related to oxidative stress and predispose cells to malignanttransformation in the oral cavity.     

Oral cancer prediction by noninvasive genetic screening

Oral squamous cell carcinomas (OSCCs) develop in genetically altered epithelium in the mucosal lining, also coined as fields, which are mostly not visible but occasionally present as white oral leukoplakia (OL) lesions. We developed a noninvasive genetic assay using next-generation sequencing (NGS) on brushed cells to detect the presence of genetically altered fields, including those that are not macroscopically visible. The assay demonstrated high accuracy in OL patients when brush samples were compared with biopsies as gold standard. In a cohort of Fanconi anemia patients, detection of mutations in prospectively collected oral brushes predicted oral cancer also when visible abnormalities were absent. We further provide insight in the molecular landscape of OL with frequent changes of TP53, FAT1 and NOTCH1. NGS analysis of noninvasively collected samples offers a highly accurate method to detect genetically altered fields in the oral cavity, and predicts development of OSCC in high-risk individuals. Noninvasive genetic screening can be employed to screen high-risk populations for cancer and precancer, map the extension of OL lesions beyond what is visible, map the oral cavity for precancerous changes even when visible abnormalities are absent, test accuracy of promising imaging modalities, monitor interventions and determine genetic progression as well as the natural history of the disease in the human patient.     

Effectiveness of a Mass Media Campaign on Oral Carcinogens and Their Effects on the Oral Cavity

Objective: To develop a mass media campaign on oral carcinogens and their effects on the oral cavity in orderto increase awareness among the general population. Methods: Documentary and public service announcementshighlighting the effects of tobacco and its products were designed and developed based on principles of behavior change. Aquestionnaire, designed to determine the knowledge, attitude and practice of people regarding oral carcinogens, was usedto conduct a baseline survey at various sites in eastern Nepal. Local television channels and radio stations broadcastedthe documentary and public service announcements. An evaluation survey was then performed to assess the effectivenessof the campaign. Results: Baseline and evaluation surveys covered 1,972 and 2,140 individuals, respectively. A thirdof the baseline population consumed quid, 22% chewing tobacco, 16% gutka (commercial preparation of arecanut,tobacco, lime and chemicals) and 25% cigarettes. Tobacco consumption differed significantly between 3 ecologic regionswith greater use in the Terai region. The knowledge prevalence regarding the oral carcinogens quid (70%), chewingtobacco (82%), gutka (58%) and cigarettes (93%) significantly increased in the evaluation population. Females weremore aware about the various tobacco products and their effects on health. More people knew about the harmful effectsof tobacco on their health and oral cavity, and had their mouth examined and the frequency of consumption of theseproducts reduced significantly after the campaign. Attitudes towards production, sale and advertisements of tobaccoalso improved significantly. Conclusions: The mass media campaign was an effective tool for increasing awarenessamong the population.     

Dilemmas of Oral Cancer Screening: An Update

Oral cancer is a global health burden with high mortality and morbidity. Advances in treatment have failedto improve the relatively poor survival rate due to late-stage diagnosis. Early detection and screening have beenshown to be effective in reducing mortality and morbidity of most common cancers. Several studies have evaluatedthe effectiveness of oral cancer screening programs but clear results were not obtained. This narrative commentaryaimed to give a critical insight into the dilemma of oral cancer screening and to suggest recommendations forfuture trends. Conventional oral examination still constitutes the gold standard screening tool for potentiallymalignant oral lesions and cancer. Interestingly, the findings of the most lasting (15-year) randomized controlledtrial on oral cancer screening using visual examination (Kerala) supported the introduction of a screeningprogram in high-risk individuals. Several screening adjuncts exist but are still not at the introduction stage.Further research to find an appropriate adjunct reliable tool for oral cancer screening is needed. In conclusion,oral cancer fulfills most of the essential principles of cancer screening but still many points need to be clarified.Therefore, there is a striking need to establish a global consortium on oral cancer screening that will overseeresearch and provide recommendations for health authorities at regular intervals.     

Saliva-Based Screening of High-Risk Human Papillomavirus Strains: Detection in Female Indonesian and Thai Dental Students

Background: Currently it is believed that human papillomaviruses (HPV) are associated with the developmentof some oral/oropharyngeal cancers. It has been suggested that these viruses influence carcinogenesis in bothsmokers and non-smokers. Data on the prevalence of HPV in healthy adults are thus needed to estimate therisk of oral/oropharyngeal cancer. The aim of this study was to assess the prevalence of oral HPV in healthyfemale adults in Indonesia and Thailand. Materials and Methods: Healthy female students from the Facultiesof Dentistry of Universitas Indonesia and Chiang Mai University were asked to participate in this pilot study.DNA was extracted from saliva specimens and screened for HPV16 and HPV18 using PCR. Results: The age,marital status and sexual experience of the subjects between the two countries were not significantly different.Eight (4%) and 4 (2%) samples were positive for HPV16 and HPV18, respectively. Fisher’s Exact test founda significant difference between HPV16 positivity in subjects who were married and had sexual intercoursebut not for HPV18. Conclusions: This study successfully detected presence of HPV16 and HPV18 DNA in anumber of saliva samples from female dental school students. Marital status, experience of sexual intercourseand safe sexual practice are related to the possibility of finding HPV DNA finding in saliva. Dentists, physiciansand other health care professionals may gain significant value from the findings of this study, which provide anunderstanding of the nature of HPV infection and its risk to patient health and disease.     

前列腺神经内分泌癌诊治1例报告

1例巨大前列腺神经内分泌癌患者应用紫杉醇＋卡铂方案全身化疗2个周期,CT检查可见前列腺肿瘤几乎完全消失。总结认为前列腺神经内分泌癌常不伴有前列腺特异性抗原（PSA）升高,对内分泌治疗无效：结合病理光镜及免疫组化法作NSE、Syn、CgA染色可明确诊断．有条件的医院可进一步施用电镜检查。采用紫杉醇＋卡铂化疗有效。