176 J Contemp Med Sci | Vol. 4, No. 3, Summer 2018: 176–178 Short Communication Introduction Head and neck cancer including oral cavity, pharynx, hypopharynx, and larynx is one of the most common malig- nancies around the world.1 Moreover, 90% of head and neck cancers have been diagnosed as squamous cell carcinoma (SCC).2 Cancer is a major cause of mortality worldwide and a lot of new cases occur every year.3 The process of oral carcinogenesis is multifactorial and multistep, and the exact sequence is almost unknown.4,5 In spite of presence of risk factors such as tobacco and alcohol in some patients, most of them ultimately are not diagnosed as malignancies. This implies that genetic has an important role in susceptibility to cancers.6 Unfortunately, despite the advanced methods in surgery, radiotherapy and chemotherapy, the prognosis of oral SCC (OSCC) is not good yet. But in case of early detection, the prognosis could be better. Biopsy followed by histopathological evaluation is gold standard for diagnosis of OSCC, but it is more helpful in late stages of disease. Biomarkers are measurable indicators which can be useful for early diagnosis of some lesions.7 The most common laboratory diagnostic procedures involve the chemical and cellular analysis of blood. Other biologic fluids like saliva are also used in diagnostic tests.8 Salivary biomarkers in oropharyngeal carcinoma are cost-effective. Saliva con- tains a wide range of components and is easy access, so patients feel more comfortable. The method is also non-inva- sive and handling is safe. There are also other advantages for using saliva instead of blood.7 Therefore, nowadays there is a tendency to use of salivary biomarkers as diagnostic and prognostic factors. The matrix metalloproteinase (MMP) family involves diverse substrates.9 They are large family of zinc-dependent endopeptidase, and they have ability to digest extracellular matrix.10 Generally, MMPs are made up of a prodomain, a catalytic domain, a hinge region, and a hemopexin domain. They are secreted from the cell or anchored to the plasma membrane. MMPs have six separated groups: Collagenases, gelatinases, stromelysins, matrilysins, membrane-type MMPs and others.11 Matrix metalloproteinase-3 (stromelysin-1) is a secretory enzyme which several growth factors regulate its expression and the process of wound healing can stimulate its secretion.12 It is expressed by keratinocytes, fibroblasts, and chondrocytes.13 It has found that overexpression of MMP3 is in association with developing malignancies including head and neck carcinomas.14,15 The P53 protein is activated after DNA damage or onco- genic signals. It has an important role in cell cycle control, DNA repair, and apoptosis.16 Altered expression and func- tional loss of P53 are common genetic changes in human malignancies.17,18 Until now, conflicting results have been obtained from the study on MMP-3 and P53 markers in OSCC patients, which could be due to differences and limitations in the immuno- histochemical and RT-PCR methods in semi- quantitative cytokine assessments. In addition, these two markers have not been evaluated simultaneously in patients with OSCC. There- fore, in this study, we decided to use a completely quantitative method to examine the levels of salivary P53 and MMP-3 in OSCC patients to find their real changes in OSCC affected patients. Among the available methods, studies can perform on the tissue blocks, blood, and other biologic fluids. Saliva is believed to be a reliable tool for diagnosis of OSCC because it is in direct contact with cancerous tissue.7 Is salivary evaluation of P53 and MMP-3 a good tool for early detection of oral squamous cell carcinoma? Shima Nafarzadeh,a Mohsen Emamgholipour,b Fatimah Bijani,c Hamed Hosseinkazemi,a Amrollah Mostafazadeh,d Oveis Khakbaz,a Fatemeh Baladi,d and Soraya Khafrie Objective Oral squamous cell carcinoma (OSCC) is one of the most common malignancies around the world. Despite the advancement in treatment methods, the prognosis is still not good. Based on clinicians’ idea, the early diagnosis of the lesion can lead to better prognosis. Some salivary biomarkers such as matrix metalloproteinase-3 and P53 may detect OSCC in early stages. In this study, we wanted to compare salivary MMP-3 and P53 levels in OSCC patients and control group. Methods Fifteen patients with OSCC (9 male and 6 female) were selected from Oral Pathology Department, Babol, Iran. Salivary MMP-3 and P53 were measured by ELISA and compared with control group. Data was analyzed by ANOVA, t-test, and Mann–Whitney. Result There was no significant differences between salivary MMP-3 and P53 concentration in patients with OSCC and healthy individuals. Conclusion Based on our findings and other similar studies, salivary MMP-3 and P53 levels might not be accurate enough to detect early stages of OSCC. But there are controversial statements about these questions. So, supplementary studies are needed to be done in future. Keywords OSCC, saliva, P53, MMP3, ELISA9 aOral Health Research Center, Institute of Health, Babol University of Medical Sciences, Babol, Iran. bStudent Research Committee, Babol University of Medical Sciences, Babol, Iran. cDepartment of Oral and Maxillofacial Pathology, Dentistry School, Babol University of Medical Sciences, Babol, Iran. dBabol University of Medical Sciences, Babol, Iran. eDepartment of Biostatistics and Epidemiology, Faculty Member of Biostatistics Sciences, Babol University of Medical Sciences, Babol, Iran. Correspondence to Mohsen Emamgholipour (email:mohsenemamgholipour@gmail.com). (Submitted: 24 June 2018 – Revised version received: 02 July 2018 – Accepted: 05 August 2018 – Published online: 26 September 2018) ISSN 2413-0516 Shima Nafarzadeh et al. 177J Contemp Med Sci | Vol. 4, No. 3, Summer 2018: 176–178 Short Communication P53 and MMP-3 in saliva of OSCC This study was designed based on the evaluation of P53 and MMP-3 in whole unstimulated saliva of patients with OSCC using ELISA method. Materials and Methods Fifteen patients with OSCC were selected from Oral and Maxillofacial Pathology Department of Dentistry Faculty of Babol University of Medical Sciences, Babol, Iran. A total of 9 patients were male and the rest of them were female. Their age range was between 51 and 89. Their diagnosis were proven by biopsy and histopathological evaluation. Invasion of dys- plastic tumoral epithelial cells into the connective tissue was essential for diagnosis. All slides were re-evaluated to confirm the diagnosis. All of them were in grades I and II. We informed the patients and asked them to participate in our study. Oral and written consents were obtained from participants. Moreover, 30 healthy individuals without any dysplastic or malignant lesions were selected as control group. Systemic diseases, smoking, chronic periodontitis, use of NSAIDs, corticosteroids, and antihistamines, history of chronic lung diseases or any treatment for malignancy, and pregnancy for women were our other exclusion criteria. Salivary specimens were collected at the morning between 9 and 12 AM. They were banned from eating, drinking, chewing gum, and smoking for at least 2 h before sampling. Saliva samples were obtained in a dry and sterile tube of 50 ml (1–5 ml for each factor). All samples were immediately centri- fuged at 4°C for 20 min at a speed of 6000 rpm to separate the cells and then were stored at −80°C until final analysis. Matrix metalloproteinase-3 and P53 salivary concentra- tion were measured by ELISA according to the manufacturer’s instructions (E1711Hu and E0907Hu, Shanghai Crystal Day Biotech Co., China). T-test and Mann–Whitney test were used for statistical analysis. Results About 9 men and 6 women participated in our study with the diagnosis of OSCC. The tongue was the most site of involvement. T-test showed no significant difference in saliva P53 con- centration among control (mean ± SD: 1011.136 ± 655.323) and OSCC (mean ± SD: 709.215 ± 454.961) groups (P = 0.171). Mann–Whitney analysis showed no significant differ- ences in saliva concentration of MMP-3 among control (mean ± SD: 45.406 ± 67.349) and SCC (mean ± SD: 35.706 ± 43.844) groups (P = 0.268). T-test showed no significant differences in saliva P53 concentration among men (mean ± SD: 755.808 ± 499.032) and women (mean ± SD: 639.326 ± 413.945) with SCC (P = 0.654). Mann–Whitney analysis showed no significant differ- ences in saliva concentration of MMP-3 among men (mean ± SD: 45.177 ± 53.002) and women (mean ± SD: 21.499 ± 22.004) with SCC (P = 0.906) (Fig. 1). Discussion Oral squamous cell carcinoma is one of the most common malignancies in head and neck region. Despite the advancement in therapeutic techniques, the prognosis has not been improved significantly. To achieve the goal, early diagnosis and treatment is essential. Nowadays, tendency to use biomarkers is increasing. Scientists are in favor with salivary biomarkers because their stimulation is safe and simple. In recent years, many researches have been done on serum and salivary biomarkers to find diagnostic and prog- nostic markers in malignancies. In the present research, we compared salivary MMP-3 and P53 levels in OSCC patients and control group. Zhang et al.19 in a meta-analysis study on Asian and European societies showed that there was no significant association between MMP-3 level and head and neck cancer risk. In contrast, in European subgroup MMP-3 polymor- phism was significantly in association with HNC risk.19 Our study took place in Iran and our result was the same as Asian subgroup in their research. About these findings we hypoth- esize that MMP-3 could be a diagnostic marker in some nations like Europeans. On the other hand, some authors stated in their article that the concentration of salivary MMP-3 level elevated in patients with oral SCC in compared to control group.20 But our examination did not show significant differences in these groups. Agha-Hosseini et al.21 in their study showed that serum and unstimulated salivary levels of MMP-3 are significantly correlated with each other and both of them increase in OSCC patients with higher stages of tumor. We did not work on patients’ serum biomarkers but there was no relation between salivary MMP-3 and risk of malignancy. Moreover, all of our patients were in low grade. Taghavi et al.22 in an immunohistochemical study on eso- phageal squamous cell carcinoma have not found a significant association between tumor and adjacent normal tissues based on P53 positive expression, but they stated that there was significant positive expression of P53 in esophageal SCC patients in comparison with healthy population. Other authors also showed that P53 antibody could be detected in the saliva of patients with OSCC and might be useful for early detection and screening.23 In other experiments overexpression of muted and unmuted P53 in patients with SCC was detected.24 Some other studies on salivary P53 level showed higher levels in OSCC patients compared to control group.25,26 Conflicting findings in other research showed that there is still no clear relationship found between P53 overexpression and known risk factors for esophageal and gastric cancers.27 However, we Fig. 1 Unstimulated whole saliva concentration of P53 and MMP-3 in patients with SCC in men and women. Data are expressed as mean ± SD. P < 0.05 is significant. 178 J Contemp Med Sci | Vol. 4, No. 3, Summer 2018: 176–178 P53 and MMP-3 in saliva of OSCC Short Communication Shima Nafarzadeh et al. also did not find significant difference in salivary P53 level in OSCC patients and control group. 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