Squamous Cell Carcinoma SCC25 Research Articles

The Immunohistochemical expressions and diagnostic values of E-cadherin and β-Catenin in suspected cervical lesion or cervical cancer

Background: Cervical lesions are the cellular changes of a pathologic process with a bacterial and viral etiology ranging from cervicitis, cervical intraepithelial neoplasm and invasive cancer. The integrity of the cervical epithelium is maintained by complex cell adhesion molecules. This work is aimed at determining the expression pattern diagnostic value of the adhesion molecules to enhance early detection of cancer. Methods: The study population was from 10 years retrospective archival tissue blocks of cervical cancer among women in Abuja metropolis from 2005 to 2015 Histopathological diagnosis made, from the 80 cases examined 20(25%) cases were cervicitis (CC), 15(18.8%) were cervical intraepithelial neoplasia (CINI), 15(18.8%) were cervical intraepithelial neoplasia (CIN2) while 30(37.3%) cases were squamous cell carcinoma SCC. They were examined immunohistochemically by Avidin Biotin Complex (ABC) method. E-Cadherin and Beta-Catenin, (cell adhesion molecules). Results: Statistically significant rate of positivity were found in the expression of β-catenin 100% among all the categories of the studied cases. β-catenin expression was significantly higher in CINI than in SCC, while in E-cadherin, therewere no statistically significant difference (p<0.05) between the distribution of high expression and low expression of E-cadherin in both CINI and SCC. In expression of E-cadherin among the cases studied, there was a significant difference (p<0.05) between rate of positivity and negativity for the expression of E-cadherin. A statistically significant rate of positivity was found in the expression of β-catenin among all the categories of the studied cases. In the expression E-cadherin, there was a gradual decrease in high expression from cervicitis to CINI, CIN2 and SCC. There was Significant (p<0.05) high degree of expression of E-cadherin in cervicitis and CINI against CIN2 and SCC, while a significant (p<0.05) low degree expression of E-cadherin in CIN2 and SCC against Cervicitis and CINI. In β-catenin expression, there as a gradual decrease in high expression from cervicitis to CIN2 and SCC expect in CINI were all had high expression. Conclusion: This study showed that most of the immunological markers examined can be used for identification of different stages involved in cancer progression, also for diagnosis of cancer, predictive prognosis and can be explore as therapy targets for cervicitis and Squamous cell carcinoma.

Pyrrolidinedione-thiazolidinone hybrid molecules with potent cytotoxic effect in squamous cell carcinoma SCC-15 cells

The hybrid heterocyclic molecules are perspective materials in the development of anticancer drugs. Here, the pyrrolidinedione-thiazolidinone hybrid molecules were designed as potent anticancer agents. This study aimed to investigate the cytotoxic effect of three derivatives 1-(4-hydroxyphenyl)-, 1-(4-chlorophenyl)- and 1-(4-bromophenyl)-3-[5-[2-chloro-3-(4-nitrophenyl)prop-2-enylidene]-4-oxo-2-thioxothiazolidine-3-yl]pyrrolidine-2,5-diones (Les-6287, Les-6294, and Les-6328, respectively), their effect on the production of the reactive oxygen species (ROS), apoptosis induction, and expression of genes - PPARγ, AHR, and NRFL2 – whose products are important in metabolism in human tongue squamous cell carcinoma cells of SCC-15 line. The results of resazurin reduction and lactate dehydrogenase (LDH) release assays proved the toxicity of the tested derivatives for the SCC-15 cells. Les-6287, Les-6294, and Les-6328 inhibited the viability of SCC-15 cells with the half-maximal effective concentration (EC50) in the range of 10.18–32.75 µM at 24 and 48 h treatment. These derivatives reduced the metabolism of SCC-15 cells with the half-maximal inhibitory concentration (IC50) of 6.72–39.85 µM at 24 and 48 h treatment. Les-6287, Les-6294, and Les-6328 reduced the metabolism of normal human keratinocytes of HaCaT line murine fibroblasts of Balb/c 3T3 line to a lesser extent. The compounds used in a range from 50 to 100 µM concentrations decreased ROS production in the SCC-15 cells. The derivatives Les-6287 and Les-6328 decreased the level of expression of mRNA of PPARγ, AHR, and NRFL2 genes in these cells at PPARγ siRNA knockdown and without it. Thus, the anticancer effect of studied hybrid pyrrolidinedione-thiazolidinones in the SCC-15 carcinoma cells is accompanied by a reduction of their metabolic activity and ROS level, and increase in caspase 3 activity. However, these changes are not the result of direct interaction of Les-6287, Les-6294, and Les-6328 with the PPARγ molecule.

The Effect of MMP-9 Inhibitors on the Biological Behavior of Human Oral Squamous Cell Carcinoma SCC15 Cell Line Through PI3K/Akt Signaling Pathway

Objective: To investigate the effect of MMP-9 inhibitor (Mki67) on the biology of human oral squamous cell carcinoma SCC15 cell line and to explore its mechanism of action through PI3K/Akt signaling pathway. Methods: SCC15 cells were extracted, and the supernatant was discarded. The cells were then rinsed twice with PBS, and 0, 2.5, 5, and 10 μL of Mki67 (50 mg/mL) were added to the culture respectively. The inhibition rate of cell proliferation was detected by 3-(4, 5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) method, and the cell migration was measured by Transwell chamber test. The cell apoptosis rate was detected by cytometry, and the p-Akt protein content in the cells of each group was determined by a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) kit. Results: The cell proliferation rates of the 2.5 μL, 5 μL, and 10 μL dose groups were all lower than the 0 μL group (P < 0.05) before treatment, and the cell proliferation rates in the 2.5 μL, 5μL, and 10μL dose groups decreased overtime (P < 0.05). After 24 h, with the increase of Mki67 concentration, the number of migration and invasion gradually decreased (P < 0.05), and the number of apoptosis gradually increased (P < 0.05); besides, the relative expression of MMP-9, PI3K, and Akt mRNA decreased gradually (P < 0.05), and the expression level of Akt mRNA was not statistically significant (P > 0.05). Conclusion: MMP-9 inhibitor (Mki67) can inhibit the proliferation and migration of SCC15 cell line and induce apoptosis, and its mechanism of action may be related to the inhibition of PI3K/Akt signaling pathway.

Urinary Bladder Cancer-Epidemiological and Histopathological Study

Urinary bladder cancer is the main reason for morbidity and mortality throughout the world. Urothelial carcinoma is a common primary tumor of urinary bladder (90%). To investigate the epidemiological and histopathological aspects in Libyan patients suffering from urinary bladder cancer, we performed a study on 39 bladder cancer biopsies from transurethral resection of bladder tumor (TURBT) and cystoscopic samples obtained from Oncology Department- Al Hawari Hospital, over the period of (2021- May 2022), Tiba Medical Laboratory (2021- July 2022), and Alnoon Medical laboratory (2019- January 2022) at Benghazi, Libya. Of the 39 cases, 34 (87.2%) were diagnosed in men, and 5 (12.8%) were found in women, the men/ women ratio being of 6.8/1. The highest incidence of bladder tumors was recorded in individuals aged between 61 and 80 years old. In this age group, there were admitted 24 patients with bladder cancer, representing 61.5%. The most common pathological types of urinary bladder carcinoma found was Invasive Urothelial Carcinoma TCC (48.72%) followed by noninvasive Urothelial Carcinoma TCC (46.15%), and Squamous Cell Carcinoma SCC in (5.13%). A case of SCC on top of Schistosomiasis was also seen. Grade III was found in 51.2% of our patients and 43.5% of them had stage II. This study has highlighted upon the importance of histopathological study in evaluating urinary bladder cancer. Cystoscopic studies and biopsies help in early detection of bladder neoplasms and they form the mainstay of the diagnosis and follow up. The objects of this work were to assess epidemiological features of the bladder cancer in Benghazi - Libya with regard to frequency, age and sex distribution. Furthermore, to describe the Histopathological features of the urinary bladder cancer, particularly Transitional Cell Carcinoma (TCC) in urinary bladder biopsies.

Biotin Transport-Targeting Polysaccharide-Modified PAMAM G3 Dendrimer as System Delivering α-Mangostin into Cancer Cells and C. elegans Worms

The natural xanthone α-mangostin (αM) exhibits a wide range of pharmacological activities, including antineoplastic and anti-nematode properties, but low water solubility and poor selectivity of the drug prevent its potential clinical use. Therefore, the targeted third-generation poly(amidoamine) dendrimer (PAMAM G3) delivery system was proposed, based on hyperbranched polymer showing good solubility, high biocompatibility and low immunogenicity. A multifunctional nanocarrier was prepared by attaching αM to the surface amine groups of dendrimer via amide bond in the ratio 5 (G32B12gh5M) or 17 (G32B10gh17M) residues per one dendrimer molecule. Twelve or ten remaining amine groups were modified by conjugation with D-glucoheptono-1,4-lactone (gh) to block the amine groups, and two biotin (B) residues as targeting moieties. The biological activity of the obtained conjugates was studied in vitro on glioma U-118 MG and squamous cell carcinoma SCC-15 cancer cells compared to normal fibroblasts (BJ), and in vivo on a model organism Caenorhabditis elegans. Dendrimer vehicle G32B12gh at concentrations up to 20 µM showed no anti-proliferative effect against tested cell lines, with a feeble cytotoxicity of the highest concentration seen only with SCC-15 cells. The attachment of αM to the vehicle significantly increased cytotoxic effect of the drug, even by 4- and 25-fold for G32B12gh5M and G32B10gh17M, respectively. A stronger inhibition of cells viability and influence on other metabolic parameters (proliferation, adhesion, ATP level and Caspase-3/7 activity) was observed for G32B10gh17M than for G32B12gh5M. Both bioconjugates were internalized efficiently into the cells. Similarly, the attachment of αM to the dendrimer vehicle increased its toxicity for C. elegans. Thus, the proposed α-mangostin delivery system allowed the drug to be more effective in the dendrimer-bound as compared to free state against both cultured the cancer cells and model organism, suggesting that this treatment is promising for anticancer as well as anti-nematode chemotherapy.

Effect of acidic culture conditions on the proliferation, apoptosis, and migration ability of human tongue squamous cell carcinoma cells and its related mechanism.

This study aims to explore the effect of acidic culture conditions on the proliferation, apoptosis, and migration ability of human tongue squamous cell carcinoma SCC15 and CAL27 cells and its potential molecular mechanism. After acidic culture for different periods, methyl thiazolyl tetrazolium (MTT) method was adop-ted to detect the cell proliferation of SCC15 and CAL27. Flow cytometry was employed to detect the apoptosis level of SCC15 and CAL27 cells. The migration ability of SCC15 and CAL27 after acidic culture was detected by scratch hea-ling test. Real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) was used to detect the mRNA expression of cyclooxygenase 2 (COX-2) and survivin in SCC15 and CAL27 cells after acidic culture. After culture for 24 h under acidic microenvironment, SCC15 and CAL27 cells grew rapidly and reached the stationary phase after adjustment for 3 days. The apoptosis levels of SCC15 and CAL27 cells decreased after acidic culture, but the most significant reduction occurred after 6 h of acidic culture. The scratch healing rates of SCC15 and CAL27 cells increased after acidic culture. The results of FQ-PCR showed that the mRNA expression levels of COX-2 and survivin in SCC15 and CAL27 cells increased after acidic culture. Extracellular acidic microenvironment can inhibit the apoptosis of tongue squamous carcinoma cells, promote their migration, and induce more adaptable and malignant tongue squamous carcinoma cells. The mechanism may be related to COX-2 and survivin and their signal pathways.

Maternally expressed 3 inhibits the biological activity of oral squamous cell carcinoma SCC25 and CAL27 cell lines

The abnormal expression of long non-coding RNA (lncRNA) maternally expressed 3 (MEG3) is closely related to several tumor diagnosis and progression, such as endometrial carcinoma and ovarian cancer. However, the role of MEG3 in oral squamous cell carcinoma (OSCC) is rarely reported. The current study aimed to evaluate the expression of lncRNA MEG3 in OSCC tissues and cell lines and its effect on the biological behavior of OSCC cell lines. The expression of lncRNA MEG3 in the OSCC tissues and cell lines was detected by reverse transcription-quantitative (RT-q) PCR. The relationship between MEG3 expression and the clinicopathologic characteristics and prognosis of patients with OSCC was analyzed. The lncRNA MEG3 overexpression plasmid and control plasmid were transfected into SCC25 and CAL27 cell lines using the lipofectin method. MTT assay was performed to detect the growth and proliferation of the cell lines. Transwell chamber test was used to detect changes in cell migration and invasion. Flow cytometry was employed to detect changes in apoptosis. Western blotting and RT-qPCR were conducted to detect the expression of the p53 gene. The expression of lncRNA MEG3 in the OSCC tissues and cell lines was significantly compared with normal tissues and cell lines, respectively. The expression level of MEG3 was related to clinical stage, lymph node metastasis, distant metastasis and survival status. Overexpression of lncRNA MEG3 inhibited the proliferation, migration, and invasion of SCC25 and CAL27 cell lines, induced apoptosis and promoted the expression of p53 gene. lncRNA MEG3 played the role of a tumor inhibitor gene and significantly inhibited the biological activity of OSCC cell lines, which may provide a novel idea for molecular targeted therapy of OSCC.

MO997INCIDENTAL SKIN CANCER AFTER KIDNEY TRANSPLANTATION: A REGIONAL POPULATION BASED-COHORT STUDY (2015-2020)

Abstract Background and Aims Long-term immunosuppression in solid organ transplant (SOT) recipients could be associated with high risk of cutaneous malignancies. Any suspicious lesion should be excised and sent for pathologic examination. Method We performed a descriptive retrospective single-center study to evaluate the incidence of all skin cancer lesions in 372 kidney transplanted recipients between January 2015 to July 2020 with at least 6 months of follow-up. Results We identified 53 recipients who developed skin diseases during post transplant period. Mean age was 63y, predominantly men (62%), caucasian race, with light phototype (I-II 95%).Mean follow-up were 16 months (45% first year, 23% second year, and 32%> 3 years). 70% of the patients were over 60y with an earlier development of the lesions. Actinic keratosis was the most frequent non- excised lesion. 27 patients were biopsied (51%) in whom 66% showed malignant skin lesions (squamous cell carcinoma-SCC: 39%; Basal cell carcinoma-BCC: 50%; melanoma 11%). The most frequent immunosuppressive treatment (IS) associated between these recipients was the combination of tacrolimus (Tac) and mycophenolate (MMF; 62%) versus tacrolimus (Tac) and everolimus scheme (EVE; 38%). Up to 18.5% had multiple and mixed carcinomas. 4 to 5 patients of them were with MMF regimen. This treatment was also more frequent among patients with SCC (33%). Conclusion

The Comparison of In Vitro Photosensitizing Efficacy of Curcumin-Loaded Liposomes Following Photodynamic Therapy on Melanoma MUG-Mel2, Squamous Cell Carcinoma SCC-25, and Normal Keratinocyte HaCaT Cells.

The research focused on the investigation of curcumin encapsulated in hydrogenated soy phosphatidylcholine liposomes and its increased photoactive properties in photodynamic therapy (PDT). The goal of this study was two-fold: to emphasize the role of a natural photoactive plant-based derivative in the liposomal formulation as an easily bioavailable, alternative photosensitizer (PS) for the use in PDT of skin malignancies. Furthermore, the goal includes to prove the decreased cytotoxicity of phototoxic agents loaded in liposomes toward normal skin cells. Research was conducted on melanoma (MugMel2), squamous cell carcinoma (SCC-25), and normal human keratinocytes (HaCaT) cell lines. The assessment of viability with MTT (3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide) evaluated cell death after exposure to blue light irradiation after 4 h of pre-incubation with free and encapsulated curcumin. Additionally, the wound healing assay, flow cytometry, and immunocytochemistry to detect apoptosis were performed. The malignant cells revealed increased phototoxicity after the therapy in comparison to normal cells. Moreover, liposome curcumin-based photodynamic therapy showed an increased ratio of apoptotic and necrotic cells. The study also demonstrated that nanocurcumin significantly decreased malignant cell motility following PDT treatment. Acquired results suggest that liposomal formulation of a poor soluble natural compound may improve photosensitizing properties of curcumin-mediated PDT treatment in skin cancers and reduce toxicity in normal keratinocytes.

Effect of miR-204-5p on the proliferation, migration, and invasion on tongue squamous cell carcinoma SCC25 cells by targeting bromodomain-containing protein 4

This study aimed to explore the target relationship between miR-204-5p and bromodomain-containing protein (BRD) 4, as well as their effects on cell proliferation, migration, and invasion in tongue squamous cell carcinoma SCC25. Real-time quantitative polymerase chain reaction (RT-qPCR) was performed to detect miR-204-5p and BRD4 expression levels in tongue squamous cell carcinoma and different cell lines. TargetScan and dual luciferase reporter assay were used to confirm the target relationship between miR-204-5p and BRD4. The effects of miR-204-5p on SCC25 cell proliferation were examined by cell counting kit (CCK) 8 assay, whereas those on SCC25 cell migration and invasion were determined by Transwell assay. RT-qPCR and Western blot were used to detect the effects of miR-204-5p mimics and inhibitors on BRD4 expression. Transwell and CCK8 assays were used to detect the effects of miR-204-5p on proliferation, migration, and invasion through BRD4 regulation. miR-204-5p was significantly downregulated in the tissues and cells of squamous cell carcinoma, and BRD4 showed the opposite result. The increase in miR-204-5p expression can inhibit the proliferation, migration, and invasion of SCC25 cells. TargetScan and luciferase test confirmed that miR-204-5p and BRD4 had a negative regulatory relationship with BRD4, respectively. Moreover, miR-204-5p mimics can inhibit BRD4 expression, and miR-204-5p inhibitors can promote BRD4 expression upregulation. When miR-204-5p and BRD4 were overexpressed in SCC25 cells, BRD4 can make up for the inhibitory effect of miR-204-5p on SCC25 cells. miR-204-5p could inhibit proliferation, migration and invasion in tongue squamous cell carcinoma SCC25 cells by targeting BRD4 gene.

The association between nonsteroidal anti-inflammatory drugs and skin cancer: Different responses in American and European populations

ObjectiveTo conduct a comprehensive systematic meta-analysis investigating the association of nonsteroidal anti-inflammatory drugs (NSAIDs) and their subtypes with skin cancer (SC) and its subclasses (basal cell carcinoma BCC; squamous cell carcinoma SCC; melanoma; nonmelanoma skin cancer NMSC) in general, American and European populations. MethodsPubMed, Embase, the Cochrane Library, the China National Knowledge Infrastructure and ClinicalTrials.gov were searched up to 24 February 2019. Pooled effect sizes and 95% confidence intervals were used to estimate associations. ResultsResults based on 26 original studies including 223,619 cases and 1,398,507 controls showed both NSAIDs and nonselective Cyclooxygenase (COX) inhibitors to be statistically significantly associated with a reduced risk of SC, BCC, SCC and NMSC but not with melanoma. Conversely, no association was observed between selective Cyclooxygenase 2 (COX-2) inhibitors and SC or its subclasses. Further subgroup analysis showed that the results analyzed for American populations were almost the same as those for the general population. For European populations, neither NSAIDs nor its subtypes correlated significantly with susceptibility to SC or its subclasses. ConclusionsThe use of NSAIDs might reduce the risk of SC, but many factors including study population, drug subtype, and disease subclass affect the significance of the association.

Effect of chloroquine on the apoptosis of tongue squamous cell carcinoma SCC25 cells induced by bleomycin and nedaplatin

<p><strong>Objective:</strong> To study the effect and mechanism of chloroquine-induced chemotherapy drugs bleomycin and nedaplatin on apoptosis of tongue squamous cell carcinoma SCC25 cells. <strong>Methods</strong>: The inhibitory effect of different drugs on the growth of SCC25 cells was screened by MTT assay. MTT, Hoechst33258 staining and FCM were used to observe the changes of morphology, cell cycle and apoptosis of SCC25 cells after different treatments. The quantitative analysis of chloroquine induced by Western blot The effect of chemotherapy drugs on the expression of Bax, Bcl-2 and NFkB apoptosis- related proteins in SCC25 cells. <strong>Results</strong>: The proliferation inhibition rate of bleomycin and nedaplatin SCC25 cells induced by chloroquine was significantly higher than that of bleomycin and nedaplatin alone (p<0.05); Hoechst33358 fluorescence staining and flow cytometry showed In the bleomycin group, the apoptotic rate of the chloroquine plus bleomycin group was significantly increased, and the apoptosis rate of the chloroquine + nedaplatin group was significantly higher than that of the nedaplatin group (p<0.05). The proportion of G0/G1 phase in the chloroquine + bleomycin group was higher than that in the bleomycin group (p<0.05), and the ratio of cells in the S phase to the G2/M phase was decreased; the same chloroquine + nedaplatin group G0/G1 The proportion of cells in the period was higher than that in the nedaplatin group (p<0.05), and the proportion of cells in the S phase and G2/M phase decreased. WB results showed that the expression levels of Bax and NFkB in chloroquine + bleomycin group were significantly higher than those in bleomycin group alone, and the expression level of Bcl-2 was lower than that in bleomycin group alone. Bax and NFkB in chloroquine + nedaplatin group The expression level was significantly higher than that of the nedaplatin group alone. The expression level of Bcl-2 was lower than that of the nedaplatin group alone, and the difference was significant (p<0.05). <strong>Conclusion:</strong> Chloroquine induced bleomycin and nedaplatin can significantly inhibit the proliferation of tongue squamous cell carcinoma SCC25 cells, and its proapoptotic effect may be related to the activation of mitochondrial apoptosis-related proteins and nuclear transcription factors.</p>

Aloe-emodin induces apoptosis in human oral squamous cell carcinoma SCC15 cells

BackgroundOral and pharyngeal cancer is the most common malignant human cancers. Chemotherapy is an effective approach for anti-oral cancer therapy, while the drug tolerance and resistance remain a problem for oral cancer patients. Aloe-emodin, rhein and physcion are classified as anthraquinones, which are the main pharmacodynamic ingredients of Rheum undulatum L.. This study was undertaken to investigate whether aloe-emodin, rhein and physcion show inhibiting growth and inducing apoptosis in oral squamous cell carcinoma SCC15 cells. We found that aloe-emodin show inhibiting growth and inducing apoptosis in oral squamous cell carcinoma SCC15 cells, we also investigated the underlying mechanisms of apoptosis induced by aloe-emodin.MethodsThiazolyl blue tetrazolium bromide (MTT) test was used to detect cell proliferation. Cell apoptosis was detected by flow cytometry. We also used western blot analysis to detect the potential mechanisms of apoptosis.ResultsAloe-emodin, rhein and physcion inhibit the proliferation of SCC15 cells and the order of inhibition level are aloe-emodin > Rhein > Physcion, the half maximal inhibitory concentrations (IC50) value of aloe-emodin was 60.90 μM at 48 h of treatment. Aloe-emodin treatment resulted in a time- and dose-dependent decrease in cell viability and increased the apoptotic cell ratio. The results of western blotting showed the expression levels of caspase-9 and caspase-3 proteins increased following aloe-emodin treatment.ConclusionsOur results revealed that aloe-emodin treatment could inhibit cell viability of SCC15 cells and the potential mechanism of inhibition might be through the induction of apoptosis by regulation of the expression levels of caspase-9 and caspase-3. This indicates that aloe-emodin may be a good agent for anti-oral cancer drug exploring.

Prevent Skin Cancer in the Society, an Evidence Based Integrated Care Approach

Introduction: In the Netherlands, skin cancer is the most common type of cancer KWF, 2016. More than 51.000 new cases of skin cancer are diagnosed every year and over 900 fatalities are reported RIVM, 2017. Skin cancer incidence rates increase dramatically: since 1990 the number of new patients with skin cancer quadrupled RIVM, 2017. This increase is much higher than in other types of cancer IKNL, 2016. It is expected that incidence rates of skin cancer will increase even further, probably with a factor 2 to 5 RIVM, 2017. Direct costs for medical treatment of skin cancer are estimated at 325 million each year 250-400 million RIVM, 2017. On top of that, sick leave and premature mortality are also important indirect costs in melanoma Brochez et al., 1999. Increasing use of more costly treatment, such as Mohs surgery and photodynamic therapy, is contributing to rising treatment costs Hollestein et al, 2017. In addition to the costs, this growing group of patients with skin cancer will represent an important burden on the limited specialized dermatological care in the Netherlands. Subsequent early assessment of skin cancer risk is necessary for timely preventive measures in the general population. Methods: 1- A systematic review of the literature on patient characteristics with a prognostic predictive value for future basal cell carcinoma, squamous cell carcinoma or melanoma such as age, gender, presages, signs and risk behaviour. 2- A Current practice analysis on care concerning risk behaviour, presages and signs of skin cancer. 3- Prognostic modelling using existing data of over 5000 cases from the Netherlands Comprehensive Cancer Organisation IKNL. 4- Developing risk profiles and implement these through internet and social media. Results: It will provide insight in early risk behaviours in the general population. It also provide knowledge of serious presages or signs enabling these patients to be seen by their GP timely. Conclusion: Basal cell carcinoma BCC, approx. 70%, squamous cell carcinoma SCC, approx. 20% and melanoma approx. 10% are the three most common types of skin cancer. Melanoma is the most lethal type, accounting for 90% of all skin cancer fatalities. The incidence of melanoma in the Netherlands is among the highest in Europe RIVM, 2017. Despite the fact that most non-melanoma skin cancers BCC and SCC can be cured, the treated lesions can be disfiguring or even lethal if not treated timely. Lessons learned: Risk profiles will enable healthcare professionals and citizens both with knowledge which persons are at high risk and should be screened and insight in risk behaviour. Limitations: Risk screening is costly, time consuming and it is to be expected that the adherence will be moderate. Suggestions for future research: Behaviour change intervention target groups in the society as implementation research

PS02.130: BRAIN METASTASIS IN ESOPHAGEAL CANCER: SINGLE CENTER EXPERIENCE

Abstract Background Multimodal therapy is the treatment of choice in patients with esophageal cancer. This therapeutic strategy has led to an improvement in disease related survival. Despite the pathological response to therapy, the recurrence rate is not negligible. Brain localization is not an elective site of metastasis of the esophageal tumors but with the new chemoradiotherapy strategies it seems to evidence an increase in incidence in the different series. Objectives of the current study is to assess brain metastases frequency, therapeutic strategy and survival. Methods This is a single centre observational retrospective study: from 2006 to 2017, at Upper G.I. Surgery Division of Verona University, 327 patients (144 squamous cell carcinoma-SCC and 183 adenocarcinoma-ADK) with locally esophageal SCC or ADK have been operated. Nineteen patients (6%) with brain metastases were identified and classified as sinchronous (< 6 months after surgery) and metachronous (> 6 months) We evaluated the timing of metastasis occurrence, the features of the primary tumour, the therapy adopted, the distribution of metastases and survival. Results Of the nineteen metastatic patients, five were SCC and 14 were ADK patients. Ten patients presented synchronous metastasis and 9 metachronous. Eleven had a single metastasis, whilst 8 presented multiple lesions. Metastases were mostly sovratentorial. 7 patients were treated with surgery + stereotactic radiosurgery (SRS), 4 with surgery + whole brain radiotherapy (WBRT), 5 with only WBRT, 2 with only SRS and 1 case was treated with palliative care. Median overall survival of the brain metastasis group was 25 months, significantly lower than non metastatic patients (132 months), local metastasis (34 months) and other distant metastasis (29 months). Conclusion Brain metastasis from oesophageal or oesophago-gastric junction represent a minority of the patients and present a poor prognosis. Despite that, an aggressive treatment for limited lesions can lead to a prolonged overall survival and should therefore be evaluated in specialised centres. Disclosure All authors have declared no conflicts of interest.

A role for the clock period circadian regulator 2 gene in regulating the clock gene network in human oral squamous cell carcinoma cells.

Clock genes are the core of the circadian rhythms in the human body and are important in regulating normal physiological functions. To date, research has indicated that the clock gene, period circadian clock 2 (PER2), is downregulated in numerous types of cancer, and that it is associated with cancer occurrence and progression via the regulation of various downstream cell cycle genes. However, it remains unclear whether the decreased expression of PER2 influences the expression of other clock genes in cancer cells. In the present study, short hairpin RNA interference was used to knockdown PER2 effectively in human oral squamous cell carcinoma SCC15 cells. Quantitative polymerase chain reaction was used to assess the mRNA expression levels of various clock genes and revealed that, following the knockdown of PER2 in SCC15 cells, the mRNA expression levels of PER3, brain and muscle ARNT-like 1, deleted in esophageal cancer (DEC)1, DEC2, cryptochrome circadian clock (CRY)2, timeless circadian clock, retinoic acid receptor-related orphan receptor-alpha and neuronal PAS domain protein 2 were significantly downregulated, while the mRNA expression levels of PER1 and nuclear receptor subfamily 1 group D member 1 were significantly upregulated. In addition, flow cytometric analysis demonstrated that proliferation was enhanced and apoptosis was reduced following PER2 knockdown in SCC15 cells (P<0.05). To the best of our knowledge, the present study is the first to report that PER2 is important for the regulation of other clock genes of the clock gene network in cancer cells. This is of great significance in elucidating the molecular function and tumor suppression mechanism of PER2.

Effect of ionizing radiation on the NKG2D ligand expression on the tumor cell SCC25 surface and the killing effect on tumor cells

Objective To investigate the effect of ionizing radiation on the expression of NKG2D ligand on the surface of oral squamous cell carcinoma cell line SCC25 and its cytotoxicity to tumor cells. Methods When SCC25 cells were cultured into logarithmic growth phase, they were randomly designed as control (without treatment) and experimental group (2 Gy ionizing radiation treatment) by drawing lots. Flow cytometry was used to detect the expressions of NKG2D ligands major histocompatibility complex class Ⅰ chain-related molecule (MIC)A, MICB, UL16 binding protein (ULBP)1 on the surface of SCC25 in the control group and the experimental group cultured for 24 h. Real-time fluorescence quantification polymerase chain reaction (RT-PCR) was used to detect the changes of NKG2D ligand mRNA expression on SCC25 cell surface after 24 h culture in the experimental group and the control group. The cells were prepared and divided into blank control group (NC), 2 Gy ionizing radiation group (R), NK1 group (target ratio was 5∶1), NK2 group (target ratio was 20∶1), NK1+ R group (target ratio was 5∶1, 2 Gy ionizing radiation), NK2+ R group (target ratio was 20∶1, 2 Gy ionizing radiation). After each group was cultured for 24 h, the killing abilities of ionizing radiation and natural killer (NK) cells to oral squamous cell carcinoma SCC25 cells were detected by CCK8. Results Flow cytometry experiment showed that, among the NKG2D ligands, the MICA fluorescence values of experimental group and control group were respectively 21.04±0.39, 22.90±0.40 (t=2.465, P=0.069), MICB fluorescence values were 27.58±0.50, 29.83±1.05 (t=1.936, P=0.125), and ULBP1 fluorescence values were 21.04±0.40, 21.78±0.50 (t=1.154, P=0.313). This indicated that after ionizing radiation on SCC25, the NKG2D ligand MICA, MICB, ULBP1 expression increased slightly, but the differences were not statistically significant. RT-PCR indicated that mRNA expressions of MICB, ULBP1 were significantly different between the control group and the experimental group (t=18.334, P=0.000; t=6.381, P=0.008). The expressions of the experimental group were respectively 6.49, 1.64 times as those of the control group. The results of CCK8 showed that, there was a significant difference in cell killing ability among NK1 group, NK2 group and NC group (F=344.600, P=0.000), suggesting that NK cells could kill tumor cells, and the higher ratio of NK cells and SCC25, the stronger killing effect. The comparison between R group and NC group showed that the difference in cell killing ability was not statistically significant (P=0.567). NK1+ R group and NK1 group were compared and the difference was not statistically significant (P=0.915). There was no significant difference between NK2+ R group and NK2 group (P=0.678). This showed that the killing effect of ionizing radiation was weak. Conclusion Ionizing radiation can increase the mRNA expression of NKG2D ligands MICB and ULBP1. This may provide a new way for tumor immunotherapy. The killing effect of ionizing radiation on cells is not obvious. It may be related to low radiation dose and only 24 h for cell culture. Key words: Radiation, ionizing; Killer cells, natural; NKG2D ligands

Hsa-miR-485-5p reverses epithelial to mesenchymal transition and promotes cisplatin-induced cell death by targeting PAK1 in oral tongue squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is currently a highly prevalent disease worldwide. Cisplatin (CDDP) is widely used for the chemotherapy of OSCC. Yet, the molecular mechanisms responsible for cisplatin resistance have not been fully elucidated. In this study, we showed that overexpression of p21 (RAC1) activated kinase 1 (PAK1) induced epithelial to mesenchymal transition (EMT) and significantly promoted the invasion and migration of oral squamous cell carcinoma SCC25 cells. Emerging evidence indicates a strong link between resistance to therapy and the induction of EMT in cancer. We showed that overexpression of PAK1 induced cisplatin resistance in SCC25 cells. ERCC1 and YAP can promote cisplatin resistance in human OSCC. We showed that ERCC1 and YAP protein were upregulated by PAK1 in SCC25 cells. We found that miR-485-5p inhibited PAK1 protein expression in the SCC25 cells. Contrary to PAK1, we demonstrated that overexpression of miR-485-5p reversed EMT and significantly inhibited invasion and migration. Moreover, its overexpression sensitized SCC25-CR cells (cisplatin-resistant cells) to cisplatin. Thus, we conclude that miR-485-5p reverses EMT and promotes cisplatin-induced cell death by targeting PAK1 in oral tongue squamous cell carcinoma. This study suggests that PAK1 plays an essential role in the progression of OSCC and it is a potential therapeutic target for OSCC.

Inhibition of the PI3K/AKT Pathway Sensitizes Oral Squamous Cell Carcinoma Cells to Anthracycline-Based Chemotherapy In Vitro.

Anthracycline-based chemotherapy, such as doxorubicin (Dox), while effective against many solid tumors, is not widely used for head and neck cancers. In this study, we evaluated the efficacy of Dox, and its derivative AD198 in human, canine, and feline oral squamous cell carcinomas cells (OSCC) in vitro. Dox and AD198 had significant an anti-proliferative effect on human, canine, and feline OSCC cells in dose-dependent manner. AD198 inhibited cell proliferation more effectively than Dox in tested OSCC cells. In the human oral squamous cell carcinoma SCC25 cells, Dox and AD198 increased the production of reactive oxygen species and subsequently increased apoptosis through activation of caspase signaling pathway. Dox and AD198 increased activation of AKT, ERK1/2, and p38 MAPK signaling pathways in tested OSCC cells by dose-dependent manner. The efficacy of Dox and AD198 treatments in inhibition of cell proliferation was increased in tested OSCC when combined with PI3K/AKT inhibitor, LY294002 treatment. Inhibition of PI3K/AKT reduced Dox- and AD198-induced activation of ERK1/2 and further increased Dox- and AD198-induced phosphorylation of p38 MAPK in OSCC. Our results suggest that the anthracycline therapies, such as Dox or AD198, can be more effective for treatment of OSCC when combined with inhibitors of the PI3K/AKT pathway. J. Cell. Biochem. 118: 2615-2624, 2017. © 2016 Wiley Periodicals, Inc.

Acacia catechu Ethanolic Seed Extract Triggers Apoptosis of SCC-25 Cells.

Background:Acacia catechu Willd (Fabaceae), commonly known as catechu, cachou, and black cutch, has been studied for its hepatoprotective, antipyretic, antidiarrheal, hypoglycemic, anti-inflammatory, immunomodulatory, antinociceptive, antimicrobial, free radical scavenging, and antioxidant activities.Objective:We evaluated the cytotoxic activity of ethanol extract of A. catechu seed (ACS) against SCC-25 human oral squamous carcinoma cell line.Methods:Cytotoxic effect of ACS extract was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, using concentrations of 0.1–1000 μg/mL for 24 h. A. catechu ethanol seed extract was treated SCC-25 cells with 25 and 50 μg/mL. At the end of treatment period, apoptotic marker gene expressions such as caspase 8, 9, Bcl-2, Bax, and cytochrome c were evaluated by semiquantitative reverse transcription-polymerase chain reaction. Morphological changes of ACS treated SCC-25 cells was evaluated by acridine orange/ethidium bromide (AO/EB) dual staining. Nuclear morphology and DNA fragmentation was evaluated by propidium iodide (PI) staining.Results:A. catechu ethanol seed extract treatment caused cytotoxicity in SCC-25 cells with an IC50 value of 100 μg/mL. Apoptotic markers caspases 8 and 9, cytochrome c, Bax gene expressions were significantly increased upon ACS extract treatment indicate the apoptosis induction in SCC-25 cells. This treatment also caused significant downregulation of Bcl-2 gene expression. Staining with AO/EB and PI shows membrane blebbing, and nuclear membrane distortion further confirms the apoptosis induction by ACS treatment in SCC-25 cells.Conclusion:The ethanol seed extracts of A. catechu was found to be cytotoxic at lower concentrations and induced apoptosis in human oral squamous carcinoma SCC-25 cells.SUMMARY Acacia catechu ethanolic seed extract contains phytochemicals such as epicatechin, rutin, and quercetinAcacia catechu seed (ACS) extract significantly (P < 0.001) inhibits the active proliferation of human oral squamous carcinoma (SCC-25) cellsACS extract treatment to SCC-25 cells significantly modulated the gene expressions pertaining to apoptosis and propidium iodide and acridine orange/ethidium bromide staining also confirm the apoptosis inductionAntiproliferative and apoptosis inducing activities of ACS extract is correlated with phytochemical contents. Abbreviations used: ACS: Acacia catechu seed extract; MTT: 3 (4,5 dimethylthiazol 2 yl) 2,5 diphenyltetrazolium bromide; DMSO: Dimethyl sulfoxide; AO/EO: Acridine orange/ethidium bromide; LC MS: Liquid chromatography mass spectrometry.