Anti-Cancer Effect of Bromelain and Its Combination with Cisplatin on HN5 Cell Line (Squamous Cell Carcinoma).

Statement of the Problem: Squamous cell carcinoma (SCC) comprises over 90% of oral malignancies. Cisplatin, as a selective chemotherapy agent to treat SCC, has many side effects despite its high effectiveness. There are some studies on the effects of bromelain derived from pineapple stems on different malignancies. Purpose: The aim of this study was to investigate the effect of bromelain alone and in combination with Cisplatin on oral squamous cell carcinoma (OSCC) and fibroblast cell lines. Materials and Method: In this interventional study, the HN5 cell line of OSCC and fibroblast cell line were treated with different concentrations of bromelain alone and in combination with cisplatin. Cell viability test was performed after 24, 48 and 72 hours using MTT (3-)4,5-dimethylthiazol-2-yl(-2,5 diphenyl tetrazolium bromide) assay. In the final stage, the drug-treated cells underwent flow cytometry to assess apoptosis patterns. Data were analyzed using SPSS 17, ANOVA (for general comparison of groups) and LSD post hoc tests (for comparison two groups). p< 0.05 was considered statistically significant. Results: The findings suggested that although bromelain showed toxic effects on HN5 cancer cells, its combination with Cisplatin resulted in little improvement in its effectiveness. Bromelain alone and in combination with Cisplatin presented cytotoxic effects against fibroblasts, which depended on the dosage and time exposure (p< 0.05). The flow cytometry results did not support the superior effect of the combination of two medications over Cisplatin alone (p> 0.05). Conclusion: According to the findings, although adding bromelain to Cisplatin reduced toxicity on normal tissues, the combination of these two drugs did not increase the anticancer effect of Cisplatin. Thus, bromelain in combination with Cisplatin is not recommended as an adjuvant drug for OSCC.

Evaluation of apoptotic effect of crocin, cisplatin, and their combination in human oral squamous cell carcinoma cell line HN5.

BACKGROUND: Squamous cell carcinoma (SCC) is the most common oral malignancy with high rate of mortality. Cisplatin, as the most effective chemotherapy drug, has side effects. Considering the studies on the use of crocin in saffron in the treatment of various malignancies, this study aimed at investigating the effects of crocin and cisplatin and their combination on SCC and fibroblast cell lines. MATERIALS AND METHODS: In this interventional study, HN5 and fibroblast cell lines were treated with different concentrations of crocin (12.5-50 µg/mL) and cisplatin (2, 4, 8, 16, and 32 µg/mL), and the cells were counted after 24, 48, and 72 h by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Data were analyzed with SPSS Version 17, and P < 0.05 was considered the level of significance. In the final stage, flow cytometry after 24 h in terms of the pattern of cell death was done. RESULTS: Both drugs had a toxic effect on malignant cells. One point was the high toxic effect of 8 µg/mL cisplatin not only on cancer cells (P < 0.001) but also on fibroblasts. However, combination with 12.5 µg/mL of crocin had the same effect on HN5 cell line, despite the less toxic effect in fibroblasts in comparison with cisplatin alone (P = 0.012). Apoptosis was the pattern of cell death showed by flow cytometry. CONCLUSION: Crocin in high concentrations can have not only significant toxicity in cancer cells but also side effects in healthy tissue. It seems that lower doses of crocin, in combination with cisplatin, besides having anticancer effect, can reduce the toxicity of cisplatin in healthy tissue.

Angiogenesis and Lymphangiogenesis in Salivary Gland Adenoid Cystic Carcinoma and Mucoepidermoid Carcinoma.

BACKGROUND AND PURPOSE: This study aimed to investigate the immunohistochemical expression of CD31 and podoplanin in order to examine angiogenesis and lymphangiogenesis, respectively in common malignant tumors of salivary glands. MATERIALS AND METHODS: Forty formalin-fixed, paraffinated blocks (20 adenoid cystic carcinoma and 20 mucoepidermoid carcinoma blocks) were selected from the medical archives of Amir A'lam Hospital of Tehran, Iran. Sections from the blocks were stained by CD31 and D2-40 markers via immunohistochemistry. Clinical and demographic information was extracted from the patients' records. FINDINGS: There was a significant difference between tumors in terms of intratumoral microvessel density (MVD) (P< 0.001), total MVD (P< 0.001), and intratumoral lymphatic vessel density (LVD) (P= 0.011). In mucoepidermoid carcinoma, intratumoral MVD and LVD were greater than peritumoral MVD and LVD (P= 0.001 and P< 0.001, respectively). In mucoepidermoid carcinoma, there was no relationship between histological grade with MVD (total, intratumoral or peritumoral) or LVD (total, intratumoral or peritumoral) (P> 0.05). A similar finding was reported with respect to the histopathological grade of adenoid cystic carcinoma (P> 0.05). CONCLUSION: The higher level of angiogenesis and lymphangiogenesis in mucoepidermoid carcinoma, specifically at the center of tumor, compared to adenoid cystic carcinoma, may be attributed to differences in the clinical behaviors and metastasis of tumors. Moreover, considering the high LVD at the center of tumor in mucoepidermoid carcinoma and infrequency of metastasis to regional lymph nodes in adenoid cystic carcinoma, it can play a significant role in metastasis to regional lymph nodes.

The Core Needle and Surgical Biopsy Concordance to Detect Estrogen, Progesterone, and Her-2 Receptors in Breast Cancer: A Comparative Study.

BACKGROUND &OBJECTIVES: Evaluation of estrogen receptor (ER), progesterone receptor (PR), and (human epidermal growth factor receptor-2) Her-2 on core needle biopsies (CNBs) is increasingly in use to diagnosis early breast cancer, but its concordance with surgical excision (SE) is not well documented. METHODS: The study included 100 formalin fixed, paraffin-embedded specimens of invasive breast carcinoma archived in Pathology Department of Cancer Institute, Tehran, Iran, from 2011 to 2014. Immunohistochemistry was applied to detect ER, PR, and Her-2. RESULTS: The current study findings indicated a significant correlation of 90% between CNB and SE specimens for ER expression. The correlation between CNB and SE specimens was estimated as 81% and 97.3% for PR and Her-2, respectively. DISCUSSION: CNB can be performed confidently to determine ER and Her-2. For PR, results obtained from CNB should be considered.

A comparative study of PCNA and Ki-67 expression in dental follicle, dentigerous cyst, unicystic ameloblastoma and ameloblastoma.

Various cell proliferation markers are used as diagnostic and prognostic tools in oral lesions. Simultaneous evaluation of these markers can increase the precision of estimation of the proliferative status of different tissues. In this study we investigated the expression of PCNA and Ki-67 as markers of cell proliferation in 15 paraffin embedded samples of each dental follicle, dentigerous cyst, unicystic ameloblastoma and ameloblastoma belonging to a total of 30 male and 30 female paients using immunohistochemistry method. Expression levels based on the intensity and the percentage of stained cells was separately analyzed for each marker with chi-square test, the results of which were significant for the two markers (P<0.05). The correlation coefficient between the two markers was found to be 0.88. A significant difference in the expression of Ki-67 and PCNA was observed in the four types of studied lesions.

Assessment of bax and bcl-2 immunoexpression in patients with oral lichen planus and oral squamous cell carcinoma.

Lichen planus (LP) is a chronic inflammatory disease of probable immune-based etiology. The pathogenesis of LP is unclear, but apoptotic changes in epidermal (epithelial) cells have been reported. Destruction of the basal cell layer is observed and many changes in cell proliferation, cell repair and cell death occur in the injured mucosal epithelium. The aim of this study was to evaluate and compare the expression of bax and bcl-2 in oral lichen planus (OLP), well differentiated oral squamous cell carcinoma (WOSCC) and normal mucosa. Sixty one paraffin-embedded biopsy including 11 cases of WOSCC, 30 cases of OLP (n=15 erosive OLP [OLP-E], n=15 reticular OLP [OLP-R]) and 20 normal mucosa were entered in our research. We used immunohistochemistry staining method for assessing bax and bcl-2 expression in epithelial layers. The percentage of stained cells was estimated in 5 randomized microscopic fields and classified as (-): 0%, (+) :< 10%, (++): 10-25%, (+++): 26-50%, (++++): > 50% positive cells. The data were analyzed with Mann-Whitney, Chi Square, and Kruskal-Wallis tests. Significant differences in bax expression were observed among OLP, WOSCC compared to normal mucosa (P=0.008). No significant difference in bax expression between OLP-E and OLP-R compared to WOSCC was seen (P>0.05). Bcl-2 was negative for all OLP and normal mucosa samples, and weak positivity was observed in WOSCC samples. According to the findings of our study, it may be possible to correlate the difference of bax and bcl-2 expression levels among the mentioned lesions to the malignant potential of OLP.

p53 and PCNA Expression in Keratocystic Odontogenic Tumors Compared with Selected Odontogenic Cysts.

p53 and PCNA expression in keratocystic odontogenic tumors compared with selected odontogenic cysts Summary: The aim of this study was to evaluate p53 and PCNA expression in different odontogenic lesions regarding their different clinical behaviors. Slices prepared from 94 paraffin-embedded tissue blocks (25 radicular cysts (RC), 23 dentigerous cysts (DC), 23 keratocystic odontogenic tumors (KCOT) and 23 calcifying cystic odontogenic tumors (CCOT)) were stained with p53 and PCNA antibodies using immunohistochemistry procedure. The highest level of p53 expression was in the basal layer of RC, and the highest level of PCNA expression was in the suprabasal layer of KCOT. The differences of p53 expression in basal and suprabasal layers as well as PCNA expression in the suprabasal layer were significant but there was no significant difference in PCNA expression in the basal layer of these lesions. The expression of p53 in the basal layer of RC was higher than in other cysts. This may be due to intensive inflammatory infiltration. Also, the high level of PCNA expression in the suprabasal layer of KCOT may justify its neoplastic nature and tendency to recurrence. KCOT and calcifying cystic odontogenic tumors did not show similar expression of studied biomarkers.

Comparative study of expression of smad3 in oral lichen planus and normal oral mucosa.

Oral lichen planus (OLP) is a chronic inflammatory disease of the oral mucosa which is considered by the World Health Organization (WHO) as a premalignant condition. One step in malignant development is so called epithelial mesenchymal transition (EMT), a process whereby epithelial cells acquire mesenchymal characteristics. A factor known to induce EMT is the transforming growth factor-ß (TGF-ß), which uses the Smad proteins as mediators for its signaling. The aim of this study was to compare the expression of Smad 3 in Oral Lichen Planus and normal oral mucosa. This descriptive analytic study was performed on 30 patients with OLP (21 women and 9 men with mean age of 45.23± 2.44 years) and 20 normal oral mucosa (14 women and 6 men with mean age of 46.95± 2.21 years). The samples were studied by immunohistochemical staining. Data were analyzed with paired T-test and Wilcoxon test by SPSS software. Expression of Smad3 in OLP samples and normal oral mucosa was different. This difference was statistically significant (P<0.001). The apparently higher expression of Smad 3 in oral lichen planus compared to normal oral mucosa might help to discuss its higher potential for malignant transition.

Relation between pulpal neuropeptides and dental caries.

INTRODUCTION: Dental pulp has neural fibers that produce neuropeptides like Substance P (SP) and calcitonin gene-related peptide (CGRP). The inflammation of dental pulp can lead to an increase amount of SP and CGRP release, especially in symptomatic irreversible pulpitis. Therefore, it can be assumed that neuropeptides have some role in the progression of inflammation of the dental pulp. The aim of this study was to determine the relation between the presence and concentration of neuropeptides in dental pulps of carious teeth caries. MATERIALS AND METHODS: For this purpose, pulpal tissues were collected from 40 teeth (20 carious and 20 intact). Pulpal samples were cultured for 72 hours. ELISA reader was used for the detection of SP and CGRP in supernatant fluids. Statistical analysis was made by Mann-Whitney U and Chi square tests. RESULTS: SP and CGRP were present in 65% and 20% of inflamed pulpal samples, respectively and 40% and 5% of normal pulpal samples, respectively. Level of SP was significantly higher in inflamed pulp samples compared to intact pulps; however, there was no statistical difference when the other groups and neuropeptides were compared. The mean concentration of SP in normal pulps was 3.4 times greater than that of CGRP; interestingly in inflamed pulps the concentration of SP was 22.3 times greater than CGRP. CONCLUSION: We can conclude that in inflamed dental pulps, the concentration of SP is higher than CGRP. It can be hypothesized that CGRP has less effect on the inflammatory changes of dental pulps.

Extramedullary plasmacytoma of the gingiva.

Extramedullary plasmacytoma is defined as neoplastic proliferation of plasma cells in soft tissue. It accounts for up to 3% of all plasma cell tumors. Approximately, 90% of extramedullary plasmacytomas are found in the head and neck region commonly affecting the nasal cavity, paranasal sinuses, tonsillar fossa, and oral cavity. Radiotherapy is the common modality of treatment with or without adjuvant chemotherapy. We report a case of extramedullary plasmacytoma of the gingiva.