Lymphonode Metastasis Research Articles

749. GENDER DIFFERENCE IN ESOPHAGEAL SQUAMOUS CELL CARCINOMA

Abstract In Japan, esophageal cancer is often more common in men, with a lower incidence in women than in men, accounting for 12% of the total number of cancers. In Japan and the West, SCC is involved in alcohol and tobacco. These habits are characteristically more common among men. Women are more likely to be diagnosed at an earlier stage than men. Therefore, we examined the prognosis, stage, and pathology of 145 men and 30 women who underwent radical surgery for esophageal cancer over the past 5 years. (Result) p stage is male: stage 0:7(5%), stage 1:35(25%), stage II: 58(40%), stage III:35(25%), stage IV:10(7%) female: Stage 0: 2 (7%), Stage 1: 12 (40%), Stage II: 6 (20%), Stage III: 9 (30%), Stage IV: 1 (3%). Significantly more men had advanced cancer. No difference was observed regarding Ly and V. Although there were no differences in location and histology, depth of invasion and lympho node metastasis were significantly enhanced in males. (table 1) Kaplanmier survival rate after 5 years was significantly better for women than for men. (Fig 1).(Conclusion) Compared to men, women had less advanced cancer at the time of discovery and a better 5-year survival rate.

Intraoperative Radioguided Localization of an Occult Neuroendocrine Tumor of the Ileum.

We report the intraoperative radioguided localization of an occult well-differentiated neuroendocrine tumor of the ileum in a patient with a known neuroendocrine lymphonodal metastasis in the mesentery. Preoperative imaging included CT, PET/CT with 68 Ga-DOTATOC, and SPECT/CT with 99m Tc-HYNICTOC. These scans revealed morphological and functional properties of the primary tumor localized in the ileum. The day after IV 99m Tc-HYNICTOC injection, the patient underwent surgery, and the target lesion was intraoperatively detected by a collimated handheld γ-probe. Postoperative pathology examination confirmed the target lesion to be a neuroendocrine tumor of the ileum.

Prognostic value of pretreatment inflammatory markers in localised prostate cancer before radical prostatectomy

PurposeThere is growing evidence of an association between inflammatory processes and cancer development and progression. In different solid tumor entities, a pronounced inflammatory response is associated with worse oncological outcome. In this study, we aim to evaluate the prognostic role of clinically established pretreatment inflammatory markers in patients with localised prostate cancer (PCa) before radical prostatectomy (RP).MethodsA total of 641 men met our inclusion criteria and were followed prospectively for a median of 2.85 years. Univariable logistic and Cox regression analysis were performed to analyse associations between preoperative inflammatory markers and tumor characteristics, and biochemical recurrence free survival (BRFS).ResultsMedian age at RP was 64 years. Gleason Score (GS) 7a (263, 41%) was the most prevalent histology, whereas high-risk PCa (≥ GS 8) was present in 156 (24%) patients. Lympho-nodal metastasis and positive surgical margin (PSM) were detected in 69 (11%) and 180 (28%) patients, respectively. No statistically relevant association could be shown between pretreatment inflammatory markers with worse pathological features like higher tumor stage or grade, nodal positive disease or PSM (for all p > 0.05). Additionally, pretreatment inflammatory markers were not associated with a shorter BRFS (p > 0.05). Known risk factors (tumor grade, tumor stage, nodal positivity and positive surgical margins) were all associated with a shorter BRFS (for all p < 0.0001).ConclusionIn this large prospective cohort, preoperative inflammatory markers were not associated with worse outcome.

Prognostic Potential of the Expression of Podoplanin (D2-40) Within Cells of Squamous Cell Carcinoma of the Larynx and Hypopharynx.

BackgroundPodoplanin (D2-20) stains immunohistochemically lymphatic vessels, regular mesothelium and tumor cells of different tumors, e.g. malignant mesothelioma or seminoma. In squamous cell carcinoma (SCC), the marker has been described as variously expressed.MethodsThis study has investigated the value of the immunohistochemical analysis for the prognostic relevance of the expression in 119 SCCs of the larynx and hypopharynx. The clinical data and documentation of follow-up for at least 5 years were available.ResultsThe collective showed the expected distribution of patient age with accentuation of the male sex and a balanced spread of tumor stages including nodal status. The immunohistochemical stain intensity (negative, weak or strong) and the distribution (equal versus focal) were evaluated. In addition, the accentuation of the staining reaction was separately examined at the border of invasion. SCCs with a strong expression of podoplanin were associated with an unfavorable prognosis. A comparison of grouped cases showed a trend emerging with borderline results (negative to weakly positive, P = 0.51; negative to strongly positive, P = 0.054; weakly positive to strongly positive, P = 0.17). The staining at the border of invasion had no statistical effect on overall survival. Multivariate survival statistics however showed that lymphonodal metastasis and a reaction with podoplanin in tumor cells are associated with significant worse prognosis.ConclusionIn summary, regardless of the exact function of podoplanin in the process of cell migration and tumor progression, an immunohistochemical identification of expression in tumor cells of SCC of the larynx and hypopharynx can give additional information about the expectable prognosis.

MRNA-Expression of KRT5 and KRT20 Defines Distinct Prognostic Subgroups of Muscle-Invasive Urothelial Bladder Cancer Correlating with Histological Variants.

Recently, muscle-invasive bladder cancer (MIBC) has been subclassified by gene expression profiling, with a substantial impact on therapy response and patient outcome. We tested whether these complex molecular subtypes of MIBC can be determined by mRNA detection of keratin 5 (KRT5) and keratin 20 (KRT20). Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) was applied to quantify gene expression of KRT5 and KRT20 using TaqMan®-based assays in 122 curatively treated MIBC patients (median age 68.0 years). Furthermore, in silico analysis of the MD Anderson Cancer Center (MDACC) cohort (GSE48277 + GSE47993) was performed. High expression of KRT5 and low expression of KRT20 were associated with significantly improved recurrence-free survival (RFS) and disease-specific survival disease specific survival (DSS: 5-year DSS for KRT5 high: 58%; 5-year DSS for KRT20 high: 29%). KRT5 and KRT20 were associated with rates of lymphovascular invasion and lymphonodal metastasis. The combination of KRT5 and KRT20 allowed identification of patients with a very poor prognosis (KRT20+/KRT5−, 5-year DSS 0%, p < 0.0001). In silico analysis of the independent MDACC cohorts revealed congruent results (5-year DSS for KRT20 low vs. high: 84% vs. 40%, p = 0.042). High KRT20-expressing tumors as well as KRT20+/KRT− tumors were significantly enriched with aggressive urothelial carcinoma variants (micropapillary, plasmacytoid, nested).

Risk factors for ovarian involvement in young and premenopausal endometrioid endometrial cancer patients

ObjectiveThis study aimed to investigate the incidence of ovarian malignant involvement in young and premenopausal endometrioid endometrial cancer and study the possible risk factors. MethodsPremenopausal patients 45 years of age or younger with endometrioid endometrial cancer treated at the OB/GYN Hospital of Fudan University between 2009 and 2013 were identified. The incidence of ovarian malignant involvement in young and premenopausal endometrioid endometrial cancer patients were calculated and the possible risk factors were investigated. ResultsA total of 144 younger (age ≤ 45, premenopausal) patients with endometrioid endometrial cancer were identified and coexisting malignant ovarian neoplasms were detected in 6 patients. Univariate analysis revealed that deeper myometrial invasion, positive lymphonode metastasis, positive LVSI, and high histologic grade (G2-G3) were associated with ovarian involvement in younger endometrial cancer patients. However, multivariate analysis revealed that only deep myometrial invasion was an independent risk factors for ovarian involvement (OR = 12.81, P = 0.046). ConclusionIn conclusions, the incidence of coexisting ovarian malignant neoplasms in young and premenopausal patients with endometrioid endometrial cancer is low, and these findings may facilitate preoperative counseling of patients and decision making at the time of surgery.

Glucose-regulated protein 94 mediates progression and metastasis of esophageal squamous cell carcinoma via mitochondrial function and the NF-kB/COX-2/VEGF axis.

Esophageal cancer is a worldwide health problem with a very poor prognosis. Therefore, new diagnostic biomarkers or therapeutic strategies for identifying and managing esophageal squamous cell carcinoma (ESCC) are urgently needed. Glucose-regulated protein 94 (GRP94) is one of major endoplasmic reticulum-stress response proteins that plays a key role in cancer progression and therapeutic responses. However, the role of GRP94 in ESCC progression and metastasis remains unclear. The tissue array results indicated that higher GRP94 expression levels were associated with lower overall survival and higher lympho-node metastasis. Silencing GRP94 (GRP94-KD) reduced cell proliferation, migration and invasion in ESCC cells. In a xenotransplantation assay, silencing GRP94 reduced cell proliferation in the zebrafish embryo. Transmission electron microscopy revealed impaired mitochondria in GRP94-KD cells, which exhibited reduced basal respiration, spare respiratory capacity and ATP production and increased oxidative damage compared with scrambled control cells. Regarding the molecular mechanism underlying the effects of GRP94 knockdown, we found that silencing GRP94 may reduce the level of NF-kB, c-Jun, p38, IL-6, vascular endothelial growth factor (VEGF), and cyclooxygenase-2 (COX-2) as well as activation of AKT and ERK. In conclusion, our results indicate that silencing GRP94 in ESCC cells suppressed cancer growth and the metastatic potential via mitochondrial functions and NF-kB/COX-2/VEGF in ESCC cells.

Clinical significance and prognostic value of Triosephosphate isomerase expression in gastric cancer.

Triosephosphate isomerase (TPI) is highly expressed in many human cancers and is involved in migration and invasion of cancer cells. However, TPI clinicopathological significance and prognostic value in gastric cancer (GC) are not yet well defined. The aim of the present work was to evaluate TPI expression in GC tissue and its prognostic value in GC patients.TPI expression was analyzed in 92 primary GC tissues and 80 adjacent normal mucosa tissues from GC patients undergoing gastrectomy by immunohistochemical analysis of tissue microarrays (TMAs). Univariate and multivariate analyses were performed to investigate TPI prognostic significance in GC patients.Immunohistochemical staining score showed that TPI expression in cancer tissues was significantly higher than in adjacent normal mucosa (P < .001). Univariate analysis revealed that TPI expression, depth of invasion, lympho node metastasis, tumor node metastasis (TNM) stage, and tumor diameter were associated with negative prognostic predictors for overall survival in GC patients (P < .05). High TPI expression represented a significant predictor of shorter survival in GC patients with positive lymphatic metastasis (P = .022) and tumor diameter >5 cm (P = .018). Cox multivariate analysis identified TPI expression, TNM stage, and tumor diameter as independent prognostic factors in GC patients.TPI expression might be considered as a novel prognostic factor to evaluate GC patients’ survival.

Rac3 Regulates Cell Invasion, Migration and EMT in Lung Adenocarcinoma through p38 MAPK Pathway.

Background: The role of Rac3 in cell proliferation in lung adenocarcinoma has been tackled in our previous study. However, the role of Rac3 in cell invasion and migration of lung adenocarcinoma is still not clear.Methods: The expression of Rac3 in lung adenocarcinoma specimens and paired noncancerous normal tissues were evaluated by immunohistochemistry. Lentivirus-mediated RNA interference (RNAi) was employed to silence Rac3 in lung adenocarcinoma cell lines A549 and H1299. A p38 MAPK inhibitor (LY2228820) was employed to inhibit activity of p38 MAPK pathway. Cell invasion and migration in vitro were examined by invasion and migration assays, respectively. PathScan® intracellular signaling array kit and western blot were employed in mechanism investigation.Results: Rac3 expression was frequently higher in lung adenocarcinoma than paired noncancerous normal tissues. Rac3 expression was an independent risk factor for lymphonode metastasis, and was associated with worse survival outcome. Silencing of Rac3 inhibited cell invasion and cell migration in lung adenocarcinoma cell lines. Knockdown of Rac3 decreased activity of p38 MAPK pathway. LY2228820, which was an important p38 MAPK inhibitor, inhibited Rac3-induced cell invasion and migration of lung adenocarcinoma. E-cadherin expression was increased and vimentin expression was decreased after silencing of Rac3 or following the treatment of LY2228820.Conclusions: Our findings suggest that Rac3 regulates cell invasion, migration and EMT via p38 MAPK pathway. Rac3 may be a potential biomarker of invasion and metastasis for lung adenocarcinoma, and knockdown of Rac3 may potentially serve as a promising therapeutic target for lung adenocarcinoma.

Analysis of the association of HOTAIR single nucleotide polymorphism (rs920778) and risk of cervical cancer

We recently demonstrated that overexpression of HOTAIR (Hox transcript antisense intergenic RNA) was associated with tumor progression and radio-resistance in human cervical cancer. Considering the single nucleotide polymorphism (SNP) rs920778 (C>T) could influence HOTAIR expression and cancer predisposition in other malignancies, we herein investigated the association between rs920778 status and cervical cancer susceptibility in a Chinese population. Using the specific TaqMan PCR assay, we genotyped rs920778 in 215 cervical cancer patients and 430 age-matched healthy controls. As shown in our data, TT genotype of rs920778 was significantly correlated with the upregulation of HOTAIR (p=0.008). Compared with the healthy control, TT genotype and T allele notably indicated a much higher risk of cervical cancer [TT genotype: odds ratio (OR)=2.186, 95% confidence interval (CI)=1.378-3.466, p=0.003; T allele: OR=1.556, 95% CI=1.221-1.981]. In addition, we also found that the TT genotype of rs920778 was correlated with advanced tumor stage (p=0.039), highly histological grade (p=0.013), lympho node metastasis (p<0.001) and positive infection of high risk HPV (p<0.001). Among the patients who underwent concurrent chemo-radiotherapy, TT genotype carriers present notably resistance to the combination of EBRT+ICBT+cisplatin (p=0.023). In conclusion, we firstly reported that TT genotype of HOTAIR rs920778 was significantly associated with the cervical cancer susceptibility. Moreover, the TT genotype of rs920778 might be a potent prognostic marker in cervical cancer patients.

Mo2013 The Efficacy of an Endoscopic Self-Expandable Metal Stent for Gastric Outlet Obstruction (GOO) By Malignancies

Gastric Outlet Obstruction (GOO) may be caused by malignant diseases of the stomach, pancreas and duodenum or by lymphonodal metastasis. GOO causes intractable nausea and vomiting, abdominal distention and deficient oral intake, resulting in progressive deterioration of the patient’s quality of life (QOL). Advances in chemotherapeutic treatment have improved the prognoses of these types of cancer and the incidence of malignant GOO has increased both at the time of diagnosis and during the course of anticancer treatment. Self-expandable metal stent (SEMS) placement has widely been evaluated as a therapeutic alternative to surgical bypass for managing GOO with minimal invasion.

Expression of MALAT-1 gene in epithelial ovarian cancer and its clinical significance

Objective To investigate the expression and clinical significance of the long non-coding RNA MALAT-1 in epithelial ovarian cancer (EOC). Methods The expressions of MALAT-1 mRNA in 29 EOC tissues and 20 normal ovarian tissues were detected by real-time fluorescence quantitative PCR. The relationships between the expression of MALAT-1 mRNA in EOC and its clinical pathological characteristics were analyzed. Results The expression of MALAT-1 mRNA in EOC tissues was lower than that in normal ovarian tissues (0.089 ± 0.090 vs 1.450 ± 1.404, t = 4.331, P< 0.001). The expression of MALAT-1 mRNA in advanced (Ⅲ-Ⅳ) stage tissues was significantly lower than that in early (Ⅰ-Ⅱ) stage tissues (0.044±0.043 vs 0.174 ± 0.095, t =-5.125, P< 0.05). The expression of MALAT-1 mRNA was significantly lower in poorly differentiated and poorly-moderately differentiated ovarian cancer than that in well differentiated ones (0.070 ± 0.091 vs 0.161 ± 0.021, t = 2.409, P= 0.023). The expression of MALAT-1 mRNA was significantly lower in tissues with the lymphonode or omental metastasis than that in tissues without invasion (0.045±0.047 vs 0.142 ± 0.102, t = 3.378, P= 0.002). Conclusions The expression of long non-coding RNA MALAT-1 is lower in EOC, which is related to tumor grade, clinical stage, lymphonode and omental metastasis. It is suggested that MALAT-1 may play an inhibition role in the progress of ovarian cancer. Key words: Ovarian neoplasms; Long non-coding RNA; MALAT-1

BRAF and NRAS Mutations are Heterogeneous and Not Mutually Exclusive in Nodular Melanoma

Inhibitors of RAF inhibit the MAPK pathway that plays an important role in the development and progression of those melanoma carrying the V600E BRAF mutation, but there’s a subset of such patients who do not respond to the therapy. Various mechanisms of drug resistance have been proposed which include the clonal heterogeneity of the tumor. We have studied a population of nodular melanoma to investigate the intratumor and intertumor heterogeneity by Laser Capture Microdissection (LCM) analysis. Our results showed that BRAF and NRAS mutations were detected in 47% and 33% of nodular melanoma, respectively, and that there is a discrepancy in mutational pattern of tumoral sample because in the 36% of patients a different mutation, in at least 1 area of the tumor, was found by LCM analysis, giving evidence of the presence of different clonal cells populations. Moreover, we found that mutations in BRAF and NRAS are not mutually exclusive because they were simultaneously present in the same tumor specimens and we observed that when the 2 different mutations were present one is a high-frequency mutation and the other is a low-frequency mutation. This was more evident in lymphonodal metastasis that resulted from wild type to mutational analysis, but showed different mutations following LCM analysis. Therefore, we believed that, when primary tumoral sample results negative to mutational analysis, if it is possible, metastases should be investigated to verify the presence of mutations. Generally, it should be searched for other mutations, in addition to BRAF V600E, so as to better understand the mechanism of drug resistance.

TES was epigenetically silenced and suppressed the epithelial–mesenchymal transition in breast cancer

The TES gene was frequently lost in breast cancer, which could inhibit tumor invasion and the formation of distant metastasis. However, the underlying mechanisms remain unknown yet. In the present study, we aimed to investigate how TES was silenced and its roles in EMT--the key step for tumor metastasis. Real-time polymerase chain reaction (PCR) and Western blot were used to detect the mRNA and protein expression of target genes; the status of TES promoter was determined by methylation-specific PCR and subsequently, DNA sequencing. Overexpression or downregulation of TES was achieved by pcDNA3.1-TES or shRNA-TES transfection. Cellular adhesion and migration were investigated by the adhesion and Transwell assays. Morphological changes of breast cancer cells were observed under the optical microscope. The Rho A activity was measured using a commercial kit, and its roles in TES-manipulated EMT were determined by real-time PCR and Western blot. The 42.3% (33/78) breast cancer tissues presented hypermethylation of the TES gene, whereas only 2 (2.6%) non-malignant cases were hypermethylated (P<0.001). Moreover, TES hypermethylation was significantly correlated with larger tumor diameter (P=0.03) and lympho node metastasis (P=0.024). In primary cultured breast cancer cells, the demethylation treatment using 5-aza-dC notably restored the expression of TES. In vitro, overexpression of TES enhanced cellular adhesion inhibited migration and suppressed EMT, while downregulation of TES impaired cellular adhesion, promoted migration, and enhanced EMT. TES overexpression also activated the Rho A signal, which is a critical factor for the effects of TES on the EMT procedure. We firstly proved that frequent loss of TES in breast cancer was caused by promoter hypermethylation, which was correlated with poor prognosis. In vitro, TES enhanced cellular adhesion, suppressed tumor migration, and inhibited EMT. Moreover, the Rho A pathway was critical for the effects of TES on EMT, which can be blocked by the Rho A inhibitor. Therefore, we propose restoration of TES as a potent strategy for breast cancer therapy.

Abstract 5308: CD9, CD63 and CD82 tetraspanins expression profile in vulvar carcinomas.

Abstract Background: Vulvar carcinomas are approximately 95 % of all vulvar tumors. This cancer presents good prognosis since early diagnosed, and survival depends of inguinal lymphonode commitment, with about 30% of patients presents lymphonode metastasis. The mechanisms involved in biological behavior of vulvar cancer remain unclear and its treatment involves physical mutilation and psychosocial factors of patients. Therefore, identification of features related with this tumors development or genesis can contribute to its knowledge and treatment. The tetraspanins are proteins that are involved in several biological processes including cell proliferation, adhesion, invasion and migration. Their reduction or lack of expression is frequently reported in metastatic and poor prognosis tumors. Our aim was to analyze CD9, CD63 and CD82 tetraspanins expression profile in vulvar carcinoma compared to normal tissue. Methods: All samples were obtained from Pathology Department of Hospital A. C. Camargo, Sao Paulo, Brazil. TMA block was constructed with 150 paraffin-embedded samples and was used in immunohistochemical assay. The tissue sections were incubated with specific antibodies against CD9, CD63 and CD82, following manufacturer's instruction. The slides were evaluated and scored semiquantitatively by assessing the staining intensity in tumor cells and the frequency of positive cells. Results: The Immunohistochemical analysis showed that all tetraspanins were expressed in tumor samples with different intensity and frequency. CD63 were expressed by a higher samples number (90%), with stronger staining both cytoplasm (79%) and nucleus (11%). Lack of CD63 expression was described in metastatic melanomas lesions, while no metastatic lesions shows higher levels of this protein. Seventy five percent of our samples expressed CD9, exclusively on cellular membrane. CD82 showed positive reaction in 45% of the samples with weakly or moderate staining, both nucleus and cytoplasm. The association of the downregulation of CD9 and/or CD82 with a worse prognosis has been shown in breast, cervical, endometrial, esophageal, head and neck, and others carcinomas. Moreover, CD82 is considered to be an important metastasis suppressor. Conclusion: Our preliminary results showed that tetraspanins may have an important role in vulvar carcinomas and CD82/CD63 nuclear expression must be better investigated. Further analyses are ongoing. Citation Format: Katia C. Carvalho, Rafael M. Rocha, Isabela W. Cunha, Fernando A. Soares. CD9, CD63 and CD82 tetraspanins expression profile in vulvar carcinomas. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 5308. doi:10.1158/1538-7445.AM2013-5308

Circulating cell-free DNA: A promising marker of regional lymphonode metastasis in breast cancer patients

We undertook the current study with untreated breast cancer to (1) role the variations in the plasma levels of cfDNA and the size distribution in early stage, (2) determine the frequency in plasma of methylation of three candidate genes, RASSF1A, MAL, and SFRP1, and (3) to determine whether detection of cfDNA variations and methylation changes in plasma might have specific clinical utility. Thirty-nine patients woman patients (median age 64 years; range, 36-90 years) who underwent surgery for primary BR and 49 healthy females' subjects (control group without any breast lesion) were evaluated. The cfDNA levels were analyzed using quantitative real-time polymerase chain reaction of β-globin. Based on the ALU repeats, the cfDNA was considered as either total (fragments of 115 bp, ALU115) or tumoral (fragments of 247 bp, ALU247). The association between the levels of the ALU247, ALU115 repeat, and ALU 247/115and the pathologic tumor characteristics was analyzed. Used methylight qPCR method, cfDNA from plasma samples of healthy donors and patients with breast cancer were evaluated for the diagnotic value of the methylation status of three genes (RASSF1A, MAL, SFRP1) frequently methylated in breast cancer. The baseline levels of cfDNA were significantly higher in the patients with cancer, and the level of ALU247 was the most accurate circulating cfDNA marker in discriminating the cancer from non-cancer subjects. A high statistical significance was found by considering the T stage and patients with regional LN metastasis positive cancers showed significantly higher cfDNA level of ALU247. Moreover, patients with methylation of at least one of the gene under investigate showed a higher quantity of cfDNA ALU115 (p< 0.0001) and ALU247 level (p< 0.0001). We observed that necrosis could be a potential source of circulating tumour-specific cfDNA ALU247; and that cfDNA ALU247 and methylated cfDNA (RASSF1A, MAL and SFRP1) are both a phenotypic feature of tumour biology.

Adenocarcinomas of the esophagogastric junction: How does surgeon’s assessment influence the fate of patients?

92 Background: Due to controversial staging systems, classifying tumors of the esophagogastric junction (EGJ) and the choice of the following surgical therapy remains a delicate affair. Methods: In this study the impact of the preoperative surgical-clinical assessment concerning assorting tumors of the EGJ was evaluated in correlation to the patient’s outcome. We analyzed clinicopathological data from 92 patients who were pre- and intraoperatively classified as distal esophageal cancer (Type I) and thus underwent esophagectomy with gastric tube reconstruction and who afterwards (final histology) in part turned out to be cardia/gastric cancers (Type II). Results: Patients with Type II cancers showed significantly more frequent lymphonodal metastasis (p=0.022) and higher recurrence rates (p=0.01), especially distant metastatic recurrence (p=0.03). Cancer-related death was also significantly higher (p=0.002) and recurrence-free survival was significantly shorter (median: 22 vs. 57 months, p=0.027). Also the thoracoabdominal approach (TA) had a favourable influence on patients’ outcome compared to the transhiatal approach (TH). Conclusions: The correct preoperative assessment of tumors of the EGJ and the appropriate surgical therapy are crucial for the outcome of the patient. Those patients with Type II cancers might experience a survival benefit by undergoing radical combined esophago (-hemi)gastrectomy with colon interposition.

Cytotoxicity of PP(Arg)(2)- and Hp(Arg)(2)-mediated photodynamic therapy and early stage of apoptosis induction in prostate carcinoma in vitro.

Porphyrin photosensitizers tend to localize in mitochondria. The depolarization of mitochondrial membrane is one of the early stages of apoptosis and Laser Scanning Fluorescence Microscopy allows to determine changes in transmembrane mitochondrial potential under influence of PDT depending on the kind of photosensitizer (PP(Arg)(2), Hp(Arg)(2)), the energy dose (5, 10, 30 and 50 J/cm(2)) and time periods (24 and 48 hours after irradiation) in the LNCaP (lymphonodal metastasis of prostate carcinoma, the androgen dependent cell line). Cyototoxicity induced by PP(Arg)(2)- and Hp(Arg)(2)-based PDT depending on energy dose and time after irradiation in prostate carcinoma is determined with MTT. Generally, it was shown that lower energy doses induce greater changes in transmembrane mitochondrial potential. Hp(Arg)(2)-based PDT was more effective causing greater mitochondrial membrane depolarization and cell viability decrease in comparison to PP(Arg)(2)-mediated PDT (in the case of maximal nontoxic photosensitizer doses used).

Molecular analysis of TP53, D9S1747, D9S162 and RPS6 in oral squamous cell carcinoma

To investigate the correlation between the frequency of molecular abnormalities of 4 loci at chromosomal 9p21 (D9S1747, D9S162, RPS6) and 17p13 (TP53) and the clinical characteristics and prognosis. The oral squamous cell carcinoma (OSCC) lesions in 71 patients were manually microdessected. Genomic DNA from these lesions and normal lymphnode tissu or peripheral blood of the same patients were extracted using the Watson's tissue kit. The loss of heterozygosity (LOH) and microsatellite instability (MI) of 17p13 and 9p21 were analyzed by PCR-page electrophoresis after DNA extraction. LOH and MI were detected in the OSCC of 48 patients (68%). The LOH and MI frequency at chromosomes 17p13 and 9P21 were 56% (35/63) and 59% (40/68) respectively. The LOH and MI frequency at 9p21 was significantly associated with WHO grading (P < 0.01) and lymphonode metastasis (P < 0.01). The LOH and MI frequency at 17p13 was significantly associated with clinical stage (P < 0.05). TP53 genetic aberration and 9p21 genetic aberration were significant prognostic factors for OSCC. The prognosis was poor in the LOH and MI positive group of chromosome 17p13 and 9p21. The frequency of LOH and MI at TP53 was the only independent factor for overall survival (P < 0.05). The LOH and MI of 17p13 and 9p21 were related to clinical stage and lymphonode metastasis. LOH of TP53 was an independent prognostic factor for OSCC.

Inhibition of cell growth induced by photosensitizer PP(Arg) 2-mediated photodynamic therapy in human breast and prostate cell lines. Part I

Photodynamic therapy can become an effective alternative method to surgery. The experiments reveal that using low photosensitizer doses and relatively low energy doses allow us to obtain effective results after PDT (to limit formation of colonies by investigated cancer cells). The prostate and breast cancer cell lines were investigated: MCF-7, a human breast cancer responsive to androgen therapy; MDA-MB231, a more aggressive human breast cancer non-responsive to androgen therapy; LNCaP, a lymphonodal metastasis of prostate carcinoma responsive to androgen therapy; DU-145, a human prostate cancer non-responsive to androgen therapy. Clonogenic assay shows that certain PP(Arg)(2) and light energy low doses stimulate the researched colony-forming cancer cells growth. Some low energy doses used during PP(Arg)(2)-mediated PDT also cause the increase in the colony-forming tumor cells. Among investigated cancer lines, MCF-7 exhibited the biggest sensibility towards PP(Arg)(2) and LNCaP the smallest one. PP(Arg)(2) based PDT is an effective method in colony growth limitation of breast cancer cell lines: MCF-7, MDA-MB231 and prostate cancer cell lines: LNCaP, DU-145.