Squamous Lines Research Articles

A dermoid cyst of the caecum

We present a case of a 39-year-old female who was found to have an adnexal mass during routine obstetric review. On ultrasound there was a 9 cm cyst superior to the right ovary, which intraoperatively was localised to the caecum. On excision, a uniloculated cyst was seen deep to the muscularis propria of the caecum, adjacent to the appendix. The cyst contained soft creamy material admixed with hair. Microscopically, the cyst had a mature stratified squamous cell lining with scattered underlying pilosebaceous elements. In the absence of ovarian and lymphoid tissue, and without mesodermal or endodermal cellular components, a diagnosis of a dermoid cyst was made.

Caffeic Acid and Metformin Inhibit Invasive Phenotype Induced by TGF-β1 in C-4I and HTB-35/SiHa Human Cervical Squamous Carcinoma Cells by Acting on Different Molecular Targets.

During the progression of epithelial cancer, the cells may lose epithelial markers and gain mesenchymal phenotype via Epithelial-Mesenchymal Transition (EMT). Such transformation of epithelial cancer cells to mesenchymal-like characteristic benefits plasticity and supports their ability to migrate. The aim of this study was to evaluate the influence of natural compound Caffeic Acid (CA) alone and in combination with antidiabetic drug Metformin (Met) on metastatic progression of two human cervical squamous cell cancer lines, C-4I and HTB-35/SiHa cells. EMT program was triggered by exposition of both epithelial cell lines to TGF-β1. Gene expression patterns related to epithelial/mesenchymal phenotype were evaluated by Real-Time PCR analysis and the protein amount was detected by western blot. The treatment of human squamous cancer cells with CA and with Met, suppressed the motility of cells and the effect depended on a particular cell line. Both compounds regulated the EMT process in C4-I and HTB-35 cells by interfering with different molecular targets. In TGF-β1-stimulated C4-I cells, CA suppressed the expression of mesenchymal transcription factor SNAI1 which resulted in enhanced expression of epithelial markers E-cadherin, Occludin and Claudin. Additionally, CA blocked MMP-9 and upregulated TIMP-1 expression, a specific inhibitor of MMP-9. In HTB-35 cells stimulated with TGF-β1, Met decreased the expression of Vimentin. By suppressing hypoxia master regulator HIF-1α, Met caused downregulation of CAIX, an enzyme involved in metastasis of aggressive malignant cells. In this study we showed that CA and Met inhibited EMT process in cancer cells via different mechanisms. However, when applied together, compounds exerted the greater effect on EMT than each compound alone. This is the first report revealing that CA alone and co-treated with Met may reverse mesenchymal phenotype of TGF-β1-treated cervical tumor cells and we believe that the use of the two small molecules may be considered as a potential therapeutic approach for metastatic cervical cancer.

Orlistat treatment induces apoptosis and arrests cell cycle in HSC-3 oral cancer cells

The present study was aimed to investigate the effect of orlistat on an oral squamous cancer line HSC-3 as well as the underlying mechanism. Cell Counting Kit-8 (CCK-8) (Dojindo, Shanghai, China) was used for the analysis of proliferation, Annexin V-FITC and propidium iodide staining for apoptosis and flow cytometry for cell cycle distribution. Western blot assay was used to determine the alteration in the expression of cyclin D1, B1, E and CDK1. The results revealed a concentration and time-dependent decrease in the proliferation of HSC-3 cells by orlistat. The viability of HSC-3 cells was reduced to 23.4 ± 2.5 and 15.7 ± 1.6% at 40 and 50 μM concentration of orlistat after 48 h. Treatment of HSC-3 cells with orlistat resulted induction of apoptosis significantly (p < 0.05). Orlistat treatment led to the increase in proportion of apoptotic cells to 38.6 ± 2.5% after 48 h compared to 0.85 ± 0.34% in the control. Analysis of cell cycle showed that population of cells in G2/M phase in the cultures treated with orlistat for 48 h increased to 59.7 ± 5% compared to 10.2 ± 1.2% in the control. However, the population of cells in the G0/G1 and S phases was subsequently decreased. The expression of cyclin D1 and E was decreased and phosphorylation of CDK1 was increased by orlistat treatment in HSC-3 cells. Thus, orlistat induces apoptosis and cell cycle arrest in G2/M phase in HSC-3 cells through decrease in expression of cyclin D1 and E and increase in phosphorylation of CDK1. Therefore, orlistat can be used for the treatment of oral squamous cancer.

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

Reflux esophagitis and its role in the pathogenesis of Barrett's metaplasia.

Reflux esophagitis damages the squamous epithelium that normally lines the esophagus, and promotes replacement of the damaged squamous lining by the intestinal metaplasia of Barrett's esophagus, the precursor of esophageal adenocarcinoma. Therefore, to prevent the development of Barrett's metaplasia and esophageal adenocarcinoma, the pathogenesis of reflux esophagitis must be understood. We have reported that reflux esophagitis, both in a rat model and in humans, develops as a cytokine-mediated inflammatory injury (i.e., cytokine sizzle), not as a caustic chemical injury (i.e., acid burn), as traditionally has been assumed. Moreover, reflux induces activation of hypoxia inducible factor (HIF)-2α, which enhances the transcriptional activity of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) causing increases in pro-inflammatory cytokines and in migration of T lymphocytes, an underlying molecular mechanism for this cytokine-mediated injury. In some individuals, reflux esophagitis heals with Barrett's metaplasia. A number of possibilities exist for the origin of the progenitor cells that give rise to this intestinal metaplasia including those of the esophagus, the proximal stomach, or the bone marrow. However, intestinal cells are not normally found in the esophagus, the stomach, or the bone marrow. Thus, the development of Barrett's intestinal metaplasia must involve some molecular reprogramming of key developmental transcription factors within the progenitor cell, a process termed transcommitment, which may be initiated by the noxious components of the gastric refluxate. This review will highlight recent studies on the pathogenesis of reflux esophagitis and on reflux-related molecular reprogramming of esophageal squamous epithelial cells in the pathogenesis of Barrett's metaplasia.

Nodal Metastasis in Neck: A Clinical Study of 60 Cases

Introduction: An enlarged neck node is frequently the first clinical manifestation of a neoplastic process in the head and neck region. The earlier the diagnosis is made, the greater the chance of improved survival. A common denominator to all head-neck malignancy is their ability to metastasize. Although metastasis is not a random event, some tumours have the propensity to extensive local invasion without metastasis whereas others metastasize early in their development.&#x0D; Objective: To evaluate the different primary sites metastasizing to cervical lymph nodes.&#x0D; Materials and Methods: A prospective observational study was carried out from July 2007 to March 2009 in the department of Otolaryngology and Head-Neck Surgery, Combined Military Hospital, Dhaka among 60 patients who had metastatic neck nodes with known primary in head-neck region and those with unknown primary were included in this study. Metastatic neck disease involving left supraclavicular or scalene nodes with a possible primary arising within the chest or abdomen were excluded from the study. All cases were advised for Fine Needle Aspiration Cytology (FNAC) of enlarged neck nodes. Tissue samples were taken from primary sites for histopathology.&#x0D; Results: Metastatic neck disease is commonly encountered in Bangladesh. A primary lesion was identified in 53(88.33%) cases however primary sites found undetected in 7(11.67%) cases. Among the primary sites 43(81.13%) cases were seen to arise from squamous lining of upper aero-digestive tract and 10(18.87%) cases were having a nonsquamous origin arising from thyroid gland (13.33%) and parotid gland (3.33%). Among the known primary sites highest incidence of metastatic neck nodes was found with carcinoma- larynx 22(36.66%) and pyriform fossa in 9(15%) cases. Forty seven (78.33%) patients were male and 13(21.67) were female with ratio being 3.6:1 of ages ranging from 20 years to 90 years.&#x0D; Conclusion: Metastatic neck disease in commonly encountered in Bangladesh. Enlarged cervical nodes in an elderly patient should always be considered as metastatic until proved otherwise.&#x0D; Journal of Armed Forces Medical College Bangladesh Vol.12(2) 2016: 56-59

Effects of sirolimus and starved culture on autophagy of human A431 cells

Objective To evaluate effects of sirolimus (a classic autophagy inducer) and starved culture on autophagy of a human cutaneous squamous cell carcinoma cell line A431. Methods Cultured A431 cells and HeLa (a human cervical carcinoma cell line) cells were both classified into 5 groups to be treated with DMEM alone (control group) , 0.1% dimethyl sulfoxide alone (DMSO group) , 20 nmol/L sirolimus (20-nmol/L sirolimus group) , 80 nmol/L sirolimus (80-nmol/L sirolimus group) , and Earle′s balanced salt solution (EBSS group) respectively. After 4-hour treatment, Western blot analysis was performed to measure the expressions of autophagy-related markers microtubule-associated protein 1 light chain 3A/3B (LC3A/B) and recombinant gamma-aminobutyric acid receptor associated protein (GABARAP) , and acridine orange staining to determine autophagy levels in these cells. Results As Western blot analysis showed, the ratio of LC3A/B -Ⅱ to LC3A/B -Ⅰ in A431 cells was similar between the control group and DMSO group (P > 0.05) , but significantly higher in the 20-nmol/L sirolimus group, 80-nmol/L sirolimus group and EBSS group than in the control group (all P < 0.05). Western blot results from HeLa cells were similar to those from A431 cells. Bivariate correlation analysis revealed that the protein expression of GABARAP was positively correlated with that of LC3A/B -Ⅰ in both HeLa cells (r = 0.869, 95% CI: 0.807-0.999, P= 0.051) and A431 cells (r= 0.837, 95% CI: -0.173-0.989, P= 0.037) , but negatively correlated with that of LC3A/B-Ⅱ in both HeLa cells (r=-0.742, 95% CI: -0.982-0.406, P = 0.042) and A431 cells (r=- 0.684, 95% CI: -0.977-0.500, P = 0.047). Acridine orange staining showed that the percentages of autophagosome-positive A431 cells and HeLa cells were significantly increased in both the 80-nmol/L sirolimus group (23.750% ± 0.260% and 33.307% ± 0.715% respectively) and EBSS group (32.450% ± 0.488% and 66.097% ± 1.141% respectively) compared with the control group (15.987% ± 0.242% and 14.117% ± 0.295%, respectively, all P < 0.05) . Conclusion The classic autophagy inducer sirolimus and starved culture can upregulate the autophagy level of A431 cells, and GABARAP may be highly correlated with LC3A/B. Key words: Autophagy; Cell line, tumor; HeLa cells; Sirolimus; Hunger; A431 cells; GABARAP

Multiple Cysts Localized to the Vulva: A Case Report

A 61-year-old woman presented to the dermatology outpatient clinic for lesions on the vulva of about 25-years duration. On physical examination the patient exhibited multiple yellow cystic papules and nodules measuring less than 1 cm each on the labia majora (Figure 1). Histopathological examination revealed a keratinous cyst lined by stratified squamous lining consistent with epidermal cysts. The patient was referred to surgeon for excision.

Role of ion transporters in the bile acid-induced esophageal injury

Barrett's esophagus (BE) is considered to be the most severe complication of gastro-esophageal reflux disease (GERD), in which the prolonged, repetitive episodes of combined acidic and biliary reflux result in the replacement of the squamous esophageal lining by columnar epithelium. Therefore, the acid-extruding mechanisms of esophageal epithelial cells (EECs) may play an important role in the defense. Our aim was to identify the presence of acid/base transporters on EECs and to investigate the effect of bile acids on their expressions and functions. Human EEC lines (CP-A and CP-D) were acutely exposed to bile acid cocktail (BAC) and the changes in intracellular pH (pHi) and Ca(2+) concentration ([Ca(2+)]i) were measured by microfluorometry. mRNA and protein expression of ion transporters was investigated by RT-PCR, Western blot, and immunohistochemistry. We have identified the presence of a Na(+)/H(+) exchanger (NHE), Na(+)/HCO3 (-) cotransporter (NBC), and a Cl(-)-dependent HCO3 (-) secretory mechanism in CP-A and CP-D cells. Acute administration of BAC stimulated HCO3 (-) secretion in both cell lines and the NHE activity in CP-D cells by an inositol triphosphate-dependent calcium release. Chronic administration of BAC to EECs increased the expression of ion transporters compared with nontreated cells. A similar expression pattern was observed in biopsy samples from BE compared with normal epithelium. We have shown that acute administration of bile acids differently alters ion transport mechanisms of EECs, whereas chronic exposure to bile acids increases the expression of acid/base transporters. We speculate that these adaptive processes of EECs represent an important mucosal defense against the bile acid-induced epithelial injury.

Spontaneous leaking craniopharyngioma causing preoperative chemical meningitis.

Spontaneous leaking craniopharyngioma causing preoperative chemical meningitis.

EXPRESS: BDNF Signaling Contributes to Oral Cancer Pain in a Preclinical Orthotopic Rodent Model.

The majority of patients with oral cancer report intense pain that is only partially managed by current analgesics. Thus, there is a strong need to study mechanisms as well as develop novel analgesics for oral cancer pain. Current study employed an orthotopic tongue cancer model with molecular and non-reflexive behavioral assays to determine possible mechanisms of oral cancer pain. Human oral squamous cell carcinoma cells line, HSC2, was injected into the tongue of male athymic mice and tumor growth was observed by day 6. Immunohistological analyses revealed a well-differentiated tumor with a localized immune response and pronounced sensory and sympathetic innervation and vascularization. The tumor expressed TMPRSS2, a protein previously reported with oral squamous cell carcinoma. ATF3 expression in trigeminal ganglia was not altered by tumor growth. Molecular characterization of the model demonstrated altered expression of several pain-related genes, out of which up-regulation of BDNF was most striking. Moreover, BDNF protein expression in trigeminal ganglia neurons was increased and inhibition of BDNF signaling with a tyrosine kinase B antagonist, ANA-12, reversed pain-like behaviors induced by the oral tumor. Oral squamous cell carcinoma tumor growth was also associated with a reduction in feeding, mechanical hypersensitivity in the face, as well as spontaneous pain behaviors as measured by the conditioned place preference test, all of which were reversed by analgesics. Interestingly, injection of HSC2 into the hindpaw did not reproduce this spectrum of pain behaviors; nor did injection of a colonic cancer cell line into the tongue. Taken together, this orthotopic oral cancer pain model reproduces the spectrum of pain reported by oral cancer patients, including higher order cognitive changes, and demonstrates that BDNF signaling constitutes a novel mechanism by which oral squamous cell carcinoma induces pain. Identification of the key role of tyrosine kinase B signaling in oral cancer pain may serve as a novel target for drug development.

Histopathology and Immunophenotyping of Congenital Lacrimal (Anlage) Fistulae.

To report the histopathological and immuno histochemical features of congenital lacrimal fistulae. Retrospective chart review of all patients who underwent a fistulectomy for congenital lacrimal fistulae, over a 3-year period from a single surgeon's (M.J.A.) database, were included in the study. A detailed lacrimal system evaluation was performed, and intraoperative findings were documented. The excised fistulae were studied using hematoxylin and eosin and periodic acid-Schiff staining. Immunophenotyping was performed using CD3, CD5, CD10, and CD20. Twelve excised fistulae of 12 patients were studied. The mean age at presentations was 5.3 years (range, 3-11 years). The deeper parts of the fistulae were lined with hypertrophied stratified squamous epithelium similar to canalicular tissue in 83.3% (10/12), reflecting its origin from the canalicular tissue, and 16.7% (2/12) originated from the lacrimal sac and were lined with columnar epithelium with areas of squamous metaplasia. The subepithelial areas showed presence of fibrosis and chronic inflammatory infiltrate. The infiltrates were lymphoplasmacytic and were positive for CD3, CD5, and CD20 and negative for CD10 immunostaining. Hypertrophied stratified squamous lining was the commonest finding. Immunophenotyping revealed features of chronic inflammatory infiltrate composed of a mixture of both T and B lymphocytes. Histopathological analysis of fistulae may be of adjunctive value in determining the origin of the anlage.

Bile Salts at Low pH Cause Dilation of Intercellular Spaces in In Vitro Stratified Primary Esophageal Cells, Possibly by Modulating Wnt Signaling

BackgroundThe presence of dilated intercellular spaces in the stratified squamous lining of the esophagus is the pathognomonic feature of reflux esophagitis secondary to gastroesophageal reflux disease (GERD). In addition to stomach acid, bile salts are major constituents of gastroesophageal refluxate. The aim of our study was to determine the effect of bile salts cocktail at different pHs on epithelial junctions in an in vitro transwell model of stratified esophageal squamous epithelium. DiscussionHuman telomerase reverse transcriptase (hTERT) immortalized primary esophageal EPC1 cells were grown on polyester transwell surfaces in calcium-enriched media. The cells exhibited gradual stratification into an 11-layered squamous epithelium over 7 days, together with epithelial barrier function as indicated by increased transepithelial electrical resistance (TEER). This stratified epithelium demonstrated well-formed tight junctions, adherens junctions, and desmosomes as visualized by immunofluorescence and electron microscopy. When exposed to short pulses of bile salts at pH 5, but not either condition alone, there was loss of stratification and decrease in TEER, concomitant with disruption of adherens junctions, tight junctions, and desmosomes, leading to the appearance of dilated intercellular spaces. At the cellular level, bile salts at pH 5 activated the Wnt pathway (indicated by increased β-catenin Ser552 phosphorylation). ConclusionIn conclusion, in our in vitro transwell model bile salts at pH 5, but not bile salts or media at pH 5 alone, modulate Wnt signaling, disrupt different junctional complexes, and cause increased permeability of stratified squamous esophageal epithelium. These changes approximate the appearance of dilated intercellular space similar to that found in GERD patients.

Sensitivity of esophageal squamous cell carcinoma cells to rapamycin can be improved by siRNA-interfered expression of p70S6K

To explore the differences in sensitivity to rapamycin of five esophageal squamous cell carcinoma cell lines with different differentiation and the changes of sensitivity of the cells after siRNA-interfered expression of p70S6K. Effects of rapamycin on proliferation of ESCC cell lines with different differentiation, EC9706, TE-1, Eca109, KYSE790 and KYSE450 cells, were investigated using cell counting kit 8 (CCK-8) assay, and according to the above results, the EC9706 cells non-sensitive to rapamycin were chosen to be transfected with p70S6K-siRNA. The changes in sensitivity of cells to rapamycin were measured in vitro and in vivo using CCK-8 kit, flow cytometry and tumor formation in nude mice. CCK-8 results showed that all the five cell line cells were sensitive to low concentration of rapamycin (<100 nmol/L), but TE-1 and EC9706 cells, which were with poor differentiation, showed resistance to high concentration of rapamycin. After EC9706 cells were treated with 50, 100, 200, 500 and 1 000 nmol/L rapamycin and p70S6K-siRNA, the proliferation rates of EC9706 cells were (48.67 ± 1.68)%, (15.45 ± 1.54)%, (14.00 ± 0.91)%, (10.97 ± 0.72)% and (2.70 ± 0.32)%, respectively, and were significantly lower than that of cells treated with 50, 100, 200, 500 and 1 000 nmol/L rapamycin and control siRNA [(74.53 ± 1.71)%, (68.27 ± 1.35)%, (71.74 ± 2.44)%, (76.23 ± 1.02)% and (80.21 ± 2.77)%] (P<0.05 for all). The results of flow cytometry showed that the ratios of cells in G1 phase of the p70S6K-siRNA, rapamycin and p70S6K-siRNA+ rapamycin groups were (53.82 ± 1.78)%, (57.87 ± 4.01)% and (73.73 ± 3.68)%, respectively, significantly higher than that in the control group (46.09 ± 2.31)% (P<0.05 for all). The results of tumor formation test in vivo showed that the inhibitory effect of rapamycin on tumor growth was stronger after the cells were transfected with p70S6K-siRNA, and the inhibition rate was 96.5%. ESCC cells with different differentiation have different sensitivity to rapamycin, and p70S6K-siRNA can improve the sensitivity of cells to rapamycin in vitro and in vivo.

Clinical features of intratarsal keratinous cysts

Intratarsal keratinous cysts (IKCs) have been frequently misdiagnosed as chalazia or epidermal cysts. We reviewed a series of cases of IKCs to identify clinical features that distinguish IKCs from other eyelid diseases. We retrospectively reviewed the medical records of 17 suspected IKC patients between January 2004 and September 2014. Seventeen patients who were clinically suspected to have IKC were enrolled. All patients presented with non-inflamed eyelid nodules fixed to the tarsus. Among them, 12 biopsy specimens were available and 11 patients (91.7%) were diagnosed with IKC, with a pathological finding of stratified squamous cell lining with keratin material. The mean patient age was 55.1 years (31-71). Six patients had a surgical history of incision or incomplete excision of the lesion, followed by recurrence. On eyelid eversion, five patients showed a white-yellow nodule, and three patients had a bluish cystic lesion. The diameter of the nodules ranged from 4 to 10 mm. The intracystic material was a milky white fluid. Ten patients underwent a complete surgical excision including partial tarsectomy and there was no recurrence. IKC can be distinguished from other intratarsal lesions by a characteristic tarsal nodule fixed to the tarsus. To prevent recurrence, complete excision with partial tarsectomy is needed.

Adenovirus-mediated IL-24 gene expression induces apoptosis in a human squamous cell carcinoma cell line COLO 16

Objective To investigate the effect of adenovirus-mediated IL-24 (Ad-IL-24) gene expression on the apoptosis in a human squamous cell carcinoma cell line COLO 16, and to explore the underlying molecular mechanism. Methods Cultured COLO 16 cells were divided into two groups to be transfected with an adenovirus vector carrying the IL-24 gene (Ad-IL-24 group) or green fluorescent protein (Ad-GFP group), while those receiving no treatment served as the control group. After culture for different durations, qPCR was performed to quantify IL-24 gene expression, methyl thiazolyl tetrazolium (MTT) assay to evaluate the proliferative activity of COLO 16 cells, flow cytometry to detect the apoptosis of COLO 16 cells, laser scanning confocal microscopy (LSCM) to observe the morphological changes of COLO 16 cells, Western blot to determine the levels of Bax and Bcl-2 proteins and to evaluate the activation of caspase-3, qPCR to determine the levels of Bax and Bcl-2 mRNAs, an immunofluorescence assay to observe the expression of Bax and Bcl-2 proteins. Statistical analysis was carried out by a two-sample t-test with the SPSS 19.0 software. Results MTT assay showed that the proliferation of COLO 16 cells in the Ad-IL-24 group was significantly inhibited as early as 4 days after the transfection; thereafter, the inhibitory effect increased in a time-dependent manner, and peaked on day 6 (P 0.05). Flow cytometry revealed that the apoptosis rate was significantly higher in the Ad-IL-24 group (13.10% ± 0.92%) than in the control group (3.69% ± 0.36%, P 0.05). LSCM demonstrated that the apoptosis of COLO 16 cells was accelerated in the Ad-IL-24 group. The immunofluorescence assay, Western blot and qPCR all showed that the mRNA and protein expressions of Bax were increased, but those of Bcl-2 were decreased in the Ad-IL-24 group compared with the Ad-GFP group and control group. Moreover, Western blot showed a protein band that could specifically bind to the anti-cleaved caspase-3 antibody in the Ad-IL-24 group, but not in the Ad-GFP group or control group. Conclusions Ad-IL-24 can induce apoptosis in human COLO 16 squamous cell carcinoma cells, probably by up-regulating Bax expression, down-regulating Bcl-2 expression, and activating caspase 3. Key words: Neoplasms, squamous cell; Interleukin-24; Apoptosis; Caspase 3; COLO 16 cell

Metastatic Neck Node - A Study of 60 Cases

Objectives: To find out the incidence, distribution, level and staging of cervical nodal metastasis.Methods: This is a retrospective study carried out in the Department of Otolaryngology Head and Neck Surgery, BSMMU and Dhaka Medical College Hospital, Dhaka, from January 2005 to December 2005. Total 60 cases were studied. The diagnosis was made by detailed history, clinical examination, investigation. Analyzed data presented by various tables, graphs and figures.Results: In this study primary lesion was identified in 88.33% cases and undetected cases were 11.7%, 81.13% were squamous lining and 18.87% non-squamous. Highest frequency of metastatic node was found with carcinoma larynx (36.66%). Male and age of 41-50 years was had highest incidence. About 43.33% had dysphagia, 38.33% had hoarseness of voice, 13.33% respiratory distress, 8.33% strider and 8.33% had referred otalgia. The disease was unilateral 81.67%, 63.33% multiple node involvement, highest involvement in level II &amp; III and stage N2 larynx was commonest site of involvement.Bangladesh J Otorhinolaryngol; April 2015; 21(1): 17-22

Effect of trypsin on the growth of a skin squamous cell carcinoma cell line A431

Objective To evaluate the effect of trypsin on the proliferation, migration and adhesion of a skin squamous cell carcinoma cell line A431. Methods Cultured A431 cells were divided into several experimental groups treated with trypsin at concentrations of 0.1, 1, 10 and 100 nmol/L for 24, 48 and 72 hours respectively, and a control group treated with DMEM complete medium only. Cell counting kit-8 (CCK8) assay was conducted to evaluate cellular proliferative activity to select the optimal concentration of trypsin. Then, some A431 cells treated with trypsin at the selected concentration for 24, 48 and 72 hours respectively (or 48 hours only) served as the experimental groups (or group), and other A431 cells treated with DMEM complete medium served as the control group. Flow cytometry was performed to assess cell cycle distribution and proliferation index, fibronectin-based adhesion assay to estimate cell adhesive capacity, and wound healing assay and Transwell assay were conducted to evaluate the migratory capacity of cells in two- and three-dimensional space. Statistical analysis was carried out by using analysis of variance, paired samples t test and chi-square test. Results The proliferative activity of A431 cells increased along with the increase of trypsin concentrations, with the strongest increasing effect observed at 100 nmol/L. After treatment with 100 nmol/L trypsin, the experimental group showed a decrease in the percentage of G1-phase cells, but an increase in the percentage of S-phase cells, proliferation index, migratory and adhesive capacity compared with the control group (all P < 0.05). Conclusion Trypsin can promote the proliferation, migration and adhesion of A431 cells. Key words: Carcinoma, squamous cell; Trypsin; Cell growth processes; Cell cycle; Cell movement; Cell line, tumor

Fistule congénitale médiane de la lèvre supérieure associée à un syndrome de Goldenhar : à propos d’un cas

We present the case of a patient with Goldenhar syndrome associated with congenital fistula in the middle of philtrum at the upper lip. The patient was supported from birth for Goldenhar syndrome. Several procedures were performed in childhood to correct a macrostomia and mandibular hypoplasia right. At 14 years old, she has a rhinoplasty to correct a complex nasal malformation. During the procedure, a hole in the upper central incisor inter-space is found. It is extended by a fistula which runs through the front palate towards the vomer rail at the base of the partition. This fistula is blind behind the vomer and can be completely resected. Pathological examination of the resection reveals a squamous lining. The median fistula of the upper lip are extremely rare : less than 30 cases reported in the literature. The unusual run of the fistula and the recent appearance of a pituitary syndrome in this patient makes us look for a continuity between it and Rathke's pocket by computed tomography imaging type. The association of Goldenhar syndrome and median congenital fistula of the upper lip was never yet described in the literature. The association with hypopituitarism was suspected with the posterior extension of the malformation, not confirmed by the scanner, but strongly suspected jointly by the neurosurgical and plastic surgery team.

Mechanism Underlying Shikonin-induced Apoptosis and Cell Cycle Arrest on SCC25 Human Tongue Squamous Cell Carcinoma Cell Line

Mechanism Underlying Shikonin-induced Apoptosis and Cell Cycle Arrest on SCC25 Human Tongue Squamous Cell Carcinoma Cell Line