bluuebunny/crossref_metadata_2025_split huggingface dataset:
Temozolomide metronomic schedule with weekly of cisplatin in metastaticsoft tissue sarcoma.
Authors: Shahid Tanweer , Agarwal Vikash , Basu Saumen , Bhattacharyya Gouri Shankar 
Published: March 2020

e21519 Background: Dacarbazine (DTIC) is the standard treatment for metastatic soft tissue sarcoma. Temozolomide (TMZ) is a potentially attractive chemotherapeutic agent for this disease because of the oral route of administration and efficacy similar to that of dacarbazine but potential for resistance exist. Metronomic administration of TMZ might be a way to overcome resistance. Cisplatin is active against melanoma and might counteract mechanisms of resistance to TMZ. Methods: We reviewed data of metastatic soft tissue sarcoma patients treated at our Institutions with cisplatin (25 mg/m2 every 7 days for 3 out of 4 weeks) plus TMZ (50 mg/m2/day from day 2 for 27days). Our practice included such scheme for patients younger than 75 years, with a performance status not worse than 2, and adequate bone marrow, liver and renal function. Assessment of response was done every 3 cycles. Toxicity was graded according to NCI-CTC. Results: From August 2007 to September 2008, 33 patients were treated with a median age of 44 years (18–74); most frequent sites of metastases were lung (18 cases), brain and lymph nodes (11 cases each); 29 patients were treated as first-line and 4 as second-line. The median number of delivered cycles was 4 (range 2- 8). Toxicity was mild, with no grade 4 event reported. Nausea and vomiting were the most frequent and severe toxic effects: grade 3 in 2 cases and grade 2 in 10 cases, respectively. Other toxicities included thrombocytopenia (1 case grade 3 and 2 cases grade 2), anemia (1 grade 3 and 2 grade 2), neutropenia, and fatigue (1 grade 3 each). Overall, 9 patients had a partial response (27.3%; 95% exact CI: 7.0–35.5) and 8 (24%) had a disease stabilization. With a median follow-up of 20 weeks (95% CI: 19–57), there were 19 progressions and a median progression-free survival of 24 weeks (95% CI: 16-nr); 9 patients died with a median survival of 50 weeks (95% CI 43-nr). Conclusions: Results obtained in clinical practice with metronomic temozolomide plus cisplatin in the treatment of patients with metastatic soft tissue sarcoma are encouraging, in light of the negative prognostic features of treated patients.
Outcome and prognostic factors in metastatic primitive neuroectodermal tumors with uniform chemotherapy protocol: A single center experience of 150 patients.
Authors: Bakhshi Sameer , Biswas Bivas , Mohanti B. K. , Shukla N. K. , Rastogi Shishir , Agarwala Sandeep , Deo S. V. S. , Sharma Dayanand , Khan Shah Alam , Thulkar Sanjay , Vishnubhatla Sreenivas 
Published: March 2020

e21522 Background: Data on metastatic primitive neuroectodermal tumor (PNET) with uniform protocol is minimal. Methods: This is single institutional patient review treated between June 2003-Nov 2011, and evaluated on intent-to-treat analysis. All patients received uniform chemotherapy (VAC/IE) as follows: neoadjuvant chemotherapy (NACT), surgery and/or radiotherapy as local treatment followed by ACT. Local treatment was offered if patient achieved CR and/or PR at both primary and metastatic site. Results: 150/374 (40%) PNET patients were metastatic with median age 15 years (range: 2–50); tumor diameter 10 cm (range: 1.8-26) and symptom duration 4 months (range: 0.1-30). Most common tumor region was pelvis 34 (23%), thorax 26 (17%) and femur in 26 (17%). Most common metastatic sites were lung only 53 (35%), bone only 35 (23%), combined bone/lung 25(17%), and bone marrow 37 (25%). Post-NACT, 9 (6%) achieved CR and 79 (53%) PR (53%) with ORR 59%. Twenty patients underwent surgery; 55 patients received radical radiotherapy following NACT. At median follow-up of 20.8 months (range: 1.6–95), 5-year EFS, OS and local control rate (LCR) were 7.6%, 15% and 37.6%, respectively. Multivariate analysis of prognostic factors in entire group and lung only metastases group is shown in the Table. Conclusions: This is the largest study of metastatic PNET from Asia and has identified unique prognostic factors. Hypoalbuminemia had inferior EFS; radical radiotherapy had inferior LCR. In group with only lung metastases, age ≤15 years and number of metastases >3 had lower EFS. In view of dismal prognosis with conventional chemotherapy in metastatic PNET, metronomic therapy may be a potential therapeutic alternative for subgroup of metastatic patients with hypoalbuminemia. [Table: see text]
Reducing invasion potential of cervical cancer cells via targeted knockdown of c-Jun.
Authors: Yee Grace Pei Chien , De Souza Paul L. , Khachigian Levon M. 
Published: March 2020

e22005 Background: Despite recent advent of vaccines for human papillomaviruses (HPV) in cervical cancer and increasing efforts to improve therapy, deaths still average 275,000 annually worldwide, with most women succumbing to recurrent or metastatic disease. The c-Jun oncogene is a subunit of the activating protein-1 (AP-1) transcription factor and is strongly expressed in cervical cancer, regulating the expression of HPV16 and 18 genes. AP-1 plays a major role in cell growth, migration and apoptosis in many cell types. This study examined the role of c-Jun in modulating HeLa proliferation, migration, apoptosis and invasion as well as susceptibility to cisplatin. Methods: Transient knockdown and over-expression of c-Jun was performed in HeLa cervical cancer cells using Dharmacon c-Jun siRNA and Origene Jun expression vector. c-Jun and downstream gene/protein expression was confirmed by western blot and real-time PCR and cells subject to proliferation, in vitro wound and matrigel dual-chamber transwell assays. Flow cytometry was used for analysis of cell cycle and apoptosis. Results: c-Jun protein and mRNA levels were reduced by c-Jun siRNA. c-Jun silencing inhibited cervical cancer cell proliferation. Significantly, c-Jun suppression dramatically reduced HeLa migration and invasion and targeted down-regulation of cyclooxygenase-2 (Cox2), interleukin-6 (IL-6), metalloproteinases (MMP)-1, -9 and -13 as well as HPV18 E6 and E7, genes highly expressed in cervical cancer and associated with metastatic growth. Direct siRNA knockdown of Cox2 in HeLa also reduced MMP1 and MMP9 expression suggesting an intermediary link. In HeLa cells over-expressing c-Jun, cell proliferation was not significantly increased but cell invasiveness was markedly enhanced in parallel with enhanced MMP-1 expression. Modulation of c-Jun expression did not interfere with susceptibility of HeLa cells to apoptosis in the presence of cisplatin. Conclusions: Reduced invasion potential of HeLa cells after c-Jun knockdown suggests a potential target in treatment of metastatic and recurrent cervical cancer. Data suggest a mechanism involving down-regulation of Cox2 and MMP-1 and -9 expression.
Tunneling nanotubes and intercellular communication: Differences between platinum-resistant and platinum-sensitive ovarian cancer.
Authors: Dickson Elizabeth Louise , Thayanithy Venugopal , Vogel Rachel Isaksson , Argenta Peter , Geller Melissa Ann , Subramanian Subbaya , Lou Emil 
Published: March 2020

e22007 Background: Preliminary evidence suggests that cell-to-cell communication may be responsible for the development of chemotherapy resistance in ovarian cancer. We propose tunneling nanotubes (TnTs) – long, thin actin-based cell extensions – as novel candidates to explain direct communication between treatment-refractory malignant ovarian cells. The purpose of this study was to investigate TnT formation between ovarian cancer cells in vitro. Methods: Using platinum-sensitive (A2780) and resistant (C200 and SKOV3, as well as ES2) ovarian cancer cell lines, we tested various conditions to assess factors affecting TnT formation. Scratch assays were utilized as a 2-dimensional simulation of ovarian cancer invasion. To assess TnTs as a conduit for transmission of therapeutic drugs between connected cells, we used doxorubicin, which auto-fluoresces in cell culture. Results: We determined that a hyperglycemic, low-serum, acidic medium stimulated TnT formation between all ovarian cancer cells studied, and more significantly, formed direct connections between A2780 to both C200 and SKOV3 cell lines. Conversely, Everolimus or Metformin decreased TnT formation in all cell lines with continuous exposure up to 96 hours; most prominently for the platinum-sensitive cell line. Time-lapse microscopy was used to assess chronologic formation of TnTs at the advancing front of the scratch wound. Cell proliferation assays were performed and confirmed the decrease in TnTs was not due to decreased cell proliferation. We directly observed fluorescing doxorubicin within the TnTs, suggesting TnTs act as a transport mechanism for cellular communication. Conclusions: TnT formation is stimulated in conditions of cellular stress similar to those experienced in vivo and results in direct connections between cells. Our data suggests that these conduits are a potential means of cellular exchange between platinum-sensitive and resistant ovarian cancer cells. Using currently available agents to target TnTs and disrupt this communication provides a novel approach to understanding and treating the problem of platinum resistance in ovarian cancer.
Use of tumor necrosis factor receptor 2 (TNFR2)/p75 and TNF-α as an antitumor strategy.
Authors: Bae Sanggyu , Sasi Sharath , Song Jin , Perepletchikov Aleksandr , Schneider Douglas , Goukassian David 
Published: March 2020

e22010 Background: TNF-α has two receptors, TNFR1/p55 and TNFR2/p75, with diverse signaling roles: p55 - predominantly cytotoxic and p75 - prosurvival and proangiogenic. Compared to WT and p55KO/p75KO, growth of tumors in p75 knockout (KO) mice was decreased >50% in Lewis lung carcinoma (LLC) and B16 melanomas (Sasi et al., Oncogene, 2012). This was associated with increased apoptosis and decreased capillary density in tumos. We hypothesized that selective blocking of p75 receptor in tumors may sensitize them to TNF induced apoptosis. Methods: p75 receptor was knockdown (KD) in LLCs using shRNA. LLC and p75KD LLCs were implanted into WT mice flanks. On day 8 post-inoculation, mouse recombinant (mr) TNF (12.5 ng/g/injection) or saline was injected x2 daily for 6 days. Tumor volumes (tV) were measured daily and tumor weights (tW) on day 16. Tumors, tubular bones and spleens were examined by 2 pathologists blindfolded to treatments. Results: Mouse weights remianed stable throughout the study in all groups. There was no difference in tV or tW between LLC minus(-) TNF and p75KD LLC-TNF tumors on day 16 (2 d days after last injection). Compared to 3 control groups, p75KD LLC+TNF showed >2-5-fold decreases in tV (p<0.001, n=8) and tW (p<0.0001, n=8). There was NO difference in tV or tW (p=ns) end of study vs before injections in p75KD LLC+TNF group whereas in 3 control groups tV and tW were increased 2.7-, 4.5- and 3.2-fold (p<0.01, p<0.0002 and p<0.0001). Pathology revealed that 1/3 of tumors in p75KD LLC+TNF group were nearly 100% necrotic. Bone marrow and spleens did not show significant TNF-induced systemic toxic effect. There was an inflammatory response in the BM and spleens of mice with necrotic tumors in +TNF group, reflected by shift of myeloid/erythroid radio in the BM and moderate increases in lymphoid follicles in the spleen. Conclusions: Our results suggest that blocking p75 expression in tumor tissue combined with administration of small exogenous doses of TNF significantly inhibits aggressive LLC tumor growth and induces nearly 100% tumor necrosis in 1/3 of the experimental group in six days, with no detectible systemic toxicity. This may represent new and effective therapy against lung neoplasms and potentially solid tumors of other origin.
Prognostic factors and survival of patients with brain metastasis (BM) from breast cancer (BC) who underwent craniotomy.
Authors: Leone Jose Pablo , Cunningham Diana E. , Lee Adrian , Bhargava Rohit , Hamilton Ronald L. , Wiener Erik , Okada Hideho , Brufsky Adam 
Published: March 2020

e22017 Background: BC is the second most frequent cause of BM after lung cancer, with metastases occurring in 10-16% of all patients. BM in patients with BC is a catastrophic event that results in poor prognosis. Identification of prognostic factors associated with breast cancer brain metastases (BCBM) could help to identify patients at risk. The aim of this study was to assess clinical characteristics, prognostic factors and survival of patients with BCBM who had craniotomy and resection in a series of patients treated with modern multimodality therapy. Methods: We analyzed 42 patients with BCBM who underwent resection. Patients were diagnosed with BC between April 1994 and May 2010. Cox proportional hazards regression was selected to describe factors associated with time to BM, survival from the date of first recurrence, and overall survival (OS). Results: Median age was 51 years (range 24-74). Median follow-up was 4.2 years (range 0.6-18.5). The mean time to BM from primary diagnosis was 49 months (range 0-206.22). Patients had a median of 2 BM with a median size of 3.25 cm. The proportion of the biological subtypes of BC was ER+/HER2- 25%, ER+/HER2+ 15%, ER-/HER2+ 30% and ER-/HER2- 30%. Brain radiotherapy was given to 28 patients, of which 10 had stereotactic radiosurgery, 7 whole brain radiation, and 11 both. Median OS from the date of primary diagnosis was 5.74 years. Median survival after diagnosis of BM was 1.33 years. In multivariate Cox regression analyses, stage was the only factor associated with shorter time to the development of BM (P=0.059), whereas age was the only factor associated with survival from the date of recurrence (P=0.027) and with OS (P=0.037). Controlling for age and stage, neither the biological subtype of cancer, the radiation modality nor the site of first recurrence showed any impact on survival. Conclusions: Stage at primary diagnosis correlated with shorter time to the development of BM, while age at diagnosis was associated with shorter survival in BCBM. None of the other clinical factors had influence on survival.
Sensitive detection of tumor nucleic acids in plasma by mutation-enriched next generation sequencing.
Authors: Marziali Andre 
Published: March 2020

e22041 Background: Next Generation DNA Sequencing (NGS) is becoming the new standard for mutational profiling of tumour tissue, due to its flexibility, speed, and decreasing cost. While generally exceptional in performance, NGS suffers from a sequencing error rate of 0.1% – 1%, largely due to amplification-induced artifacts in its workflow. While this does not constitute a significant problem in application of NGS to sequencing of tumour tissue, it makes NGS impractical as a method to search for low abundance mutation signatures in plasma samples. Numerous publications have shown the presence of tumour signatures in the cell-free DNA (cfDNA) circulating in plasma, but concordance between the tumour signature and the plasma signature has been limited. This is likely due to limitations in the detection technologies used to search for cfDNA in plasma. To maximize concordance between plasma and tissue, it will be essential that sensitivities reaching 0.01% and below (as little as a single tumour mutant allele per sample) be achieved, and ideally that multiple mutational hot spots be analysed to maximize the chance of detection. Current technologies are incapable of such sensitivity over a large number of mutation loci. Methods: We have developed a novel electrophoretic method that can enrich nucleic acid samples over 1,000,000-fold for up to 100 somatic mutations, enabling reliable profiling of samples containing as little as 0.01% mutant. By enriching nucleic acid samples for specific targets prior to amplification and sequencing, we enable the use of NGS in plasma-based mutation detection and profiling. Results: We present technical and clinical data demonstrating highly sensitive multiplexed mutation detection in plasma and tissue samples, demonstrating 0.01% sensitivity over 45 somatic mutations per sample. Conclusions: We have demonstrated a novel somatic mutation enrichment methodology that allows DNA sequencing to work beyond its usual limit of detection to accurately profile solid tumours by detecting their mutation signature in plasma, even when the tumour DNA is present in plasma at abundances below 0.01%.
Prognostic value of CDH2 and CDT2 expression levels in the peripheral blood (PB) specimens of patients with Ewing family of tumors (EFTs).
Authors: Przybyl Joanna , Debiec-Rychter Maria , Switaj Tomasz , Kosela Hanna Melania , Falkowski Slawomir , Wiater Katarzyna , Lugowska Iwona , Szumera-Cieckiewicz Anna , Ptaszynski Konrad , Siedlecki Janusz A. , Rutkowski Piotr 
Published: March 2020

e22046 Background: EFTs are characterized by oncogenic EWSR1-FLI1 fusion gene, which can be detected in 85-90% of tumors and in 20-70% of peripheral blood specimens. It has been previously reported that downregulation of CDH2 (encoding N-cadherin) and overexpression of CDT2 [also known as DTL, encoding denticleless homolog (Drosophila)]in EFTs tumors are associated with poor prognosis. The aim of the study was to evaluate the expression levels of these markers in the circulating tumor cells and assess their utility as prognostic markers in EFTs. Methods: 10mL of PB was collected from 24 untreated adult EFTs patients at the diagnosis (period: 2009-2011, median age 30 years). 9 blood specimens from healthy individuals, and 5 untreated frozen EFTs tumor samples were used as controls. 8 EFTs patients presented metastatic disease (M1) at the diagnosis and 5 patients died during the follow-up period. Median follow-up was 22 months. Quantitative reverse transcription PCR (qRT-PCR) for CDH2 and CDT2 expression were performed in triplicate using the TaqMan Gene Expression Assays (Applied Biosystems). Mean expression level of 2 reference genes PSMC4 and EIF2B1 served as endogenous control. The fold change was calculated using ddCt method with pooled healthy individuals as calibrator. Results: At least 2-fold decrease (range 2.11-16.35) of CDH2 expression level in PB has been observed in 79% (n=19) of EFTs patients (p=0.04) as compared to healthy control. In general, CDH2 expression level in PB of EFTs patients was 3.08-fold lower compared with healthy individuals (p=0.07). All of the patients who had M1 at diagnosis (n=8) and who died during the follow-up (n=5) had CDH2 gene underexpression. CDT2 overexpression (FC range 3.28-6.97) has been observed in 17% (n=4) of EFTs patients (p=0.01). Three of these patients had M1 at diagnosis and two of these patients died during the follow-up. PB specimens presenting CDT2 overexpression clustered together with the tumor specimens. Conclusions: Abnormalities in the expression of CDH2 and CDT2 genes in PB specimens are associated with aggressive clinical course of EFTs. Diagnostic and prognostic significance of these biomarkers warrants further studies.
A potential role for myosin light chain kinase in lung adenocarcinoma tumorigenesis.
Authors: Borowicz Stanley John , Tauler Jordi , Rizzo Alicia , Garcia Joe G. N. 
Published: March 2020

e22051 Background: Treatment of non-small cell lung cancer remains a clinical challenge despite the migration from cytotoxic chemotherapies to targeted agents. Myosin light chain kinase (MLCK) is a protein involved in phosphorylation of regulatory chains of myosin, thereby regulating the contraction of myosin II. The 210 KDa non-muscle MLCK isoform (nmMLCK), expressed in endothelial cells, regulates cell shape and motility, and increases EGFR-mediated proliferation of hepatocytes. Furthermore, inhibitors of MLCK such as ML-7 and ML-9 have been shown to decrease invasiveness of pancreatic prostate cancer cell lines. Clinically, levels of MLCK gene (MYLK) mRNA correlate with increased risk of disease recurrence and development of distant metastasis in lung cancer. These data suggest a potential role for MLCK in lung cancer tumorigenesis and metastasis. Methods: H-23 and H-441 human lung adenocarcinoma cell lines were treated for 12hrs with MLCK chemical inhibitor, ML-7 (20uM), and harvested at 24hr, 48hr, and 72hr for MTS Assay. Cell proliferation was assessed with MTS Assay per manufacturer protocol (Promega). H-23 cells were transiently transfected with a FLAG-tagged (WT) nmMLCK overexpression vector using Fugene Transfection Kit per manufacturer protocol (Promega). Results: While no significant difference in proliferation was noted in A-549 cells, both H-23 and H-441 cells showed a decrease in cell proliferation with inhibition of MLCK. H-23 cells also showed an increase in proliferation when transiently transfected with an MLCK-overexpression vector. Conclusions: Inhibition of endogenous nmMLCK decreases H-23 and H-441 proliferation. Conversely, an increase in nmMLCK expression in vitro increases cell proliferation in H23 cells. These results suggest that MLCK/MYLK may participate in regulation of lung adenocarcinoma cell proliferation.
Triggering receptor expressed on myeloid cells-2 (TREM-2) elicited by lung cancer cells to facilitate tumor immune evasion.
Authors: Yao Yinan , Li Hequan , Wang Yuehong , Zhou Jianying 
Published: March 2020

e22054 Background: More recent evidences have argued that negative regulatory mechanism that serve to impede ongoing immune responses, which gain in interest as a complementary strategy for cancer therapy. Triggering receptor expressed on myeloid cells-2 (TREM-2) restrains phagocytosis of both dendritic cells (DCs) and macrophages(MΦs), and reduces release of inflammatory cytokines through DAP12-mediated inhibitory signal, prompting us to investigate the role of TREM-2 involved in immune dysfunction among tumor-bearing host. Methods: In lung cancer patients, monocytes of peripheral blood were analyzed by flow cytometry and MΦs around tumour cells in lung tissue were detected by immunehistochemistry. Then we prepared conditional medium containing supernatant of 3LL cells mimicing tumor microenvironment for DC and MΦ derivation, and established an orthotopic lung cancer-bearing mice model to explore the further mechanism. Results: It was showed that TREM-2 expression on monocytes was up-regulated in lung cancer patients compared with that of healthy controls, and that TREM-2 levels of macrophages had a positive correlation with TNM stage. Moreover, reduction of tumor burden, by operation or chemotherapy, led to obvious decline of TREM-2 expression. The percentage of TREM-2+DC in the murine lung signiﬁcantly increased after tumor-bearing, and such DCs preformed lower MHCII-Ia, CD86 expression, and IL-12 production, but higher IL-10 secretion. In vitro, more TREM-2+DCs and TREM-2+MΦs were harvested from ‘conditional’ medium instead of ‘normal’ medium. Also, TREM-2+DCs rather than TREM-2- DCs markedly inhibited proliferation of T cells, which could be partially abolished by anti-TREM-2 mAb. Conclusions: TREM-2 acts as a negative immune regulator in promoting lung cancer progress. Our study provides new mechanism for tumor immune escape and inhibitory checkpoint for immunotherapy.