scholarly journals O13 Acid ceramidase: a potential biomarker for locally advanced rectal cancer?

2021 ◽  
Vol 108 (Supplement_5) ◽  
Author(s):  
Rachael Elizabeth Clifford ◽  
Naren Govindarajah ◽  
David Bowden ◽  
Paul Sutton ◽  
Jason Parsons ◽  
...  
Keyword(s):  
Rectal Cancer ◽  
Cell Lines ◽  
Locally Advanced ◽  
Underlying Mechanism ◽  
Acid Ceramidase ◽  
Clinical Role ◽  
Clonogenic Assays ◽  
Potential Biomarker ◽  
Optimal Drug

Abstract Introduction We have previously utilized proteomic and immuno-histochemical data to validate that high levels of acid ceramidase (AC) expression confers poorer neoadjuvant response in rectal cancer. Biological (siRNA, plasmid and CRISPR) AC manipulation altered radiosensitivity in-vitro. We aimed to assess the radiosensitising effect of pharmacological AC inhibitions and elucidate the potential underlying mechanism. Methods Optimal drug dosing was achieved using ELISA activity assays in multiple colorectal cancer cell lines (HCT116, HT29, LIM1215). Carmofur and a novel small molecular inhibitor (LCL521) were used as pharmacological inhibitors. Standard clonogenic assays assessed cell survival following increasing irradiation (2 D), volume change in 3 D spheroids and cell viability in patient derived organoids. Annexin V/PI staining was used to determine apoptosis. Results Carmofur clonogenic assays demonstrated reduced colony formation efficiency (CFE) and improved radiosensitivity across cell lines. HCT116 showed 0.438(CFE) control v 0.183(CFE) carmofur at 1 Gy, 0.140(CFE) control v 0.076(CFE) at 2 Gy (P = 0.000563). LCL521 dosing improved radiosensitivity in spheroid models. HCT116 volume day-15 2.36x10-5mm v control 4.15x10-5mm. siRNA-AC demonstrated increased apoptosis across time points compared to NT control (P = 0.035), and increased poly-ADP ribose polymerase-1 (PARP-1) cleavage in a p53-dependent process. Conclusion Initial work demonstrates that pharmacological inhibition of AC produces comparative radiosensitizing effects in these cell lines and cancer models. siRNA-AC increases apoptosis, suggestive of a potential underlying mechanism. This work further solidifies AC as a potential biomarker, however further recapitulation in more complex models and ultimately in-vivo is required to establish a translatable clinical role.

Acid Ceramidase As A Potential Biomarker For Locally Advanced Rectal Cancer; Is Apoptosis The Mechanism?

2019 ◽  
Vol 45 (11) ◽  
pp. 2214
Author(s):  
Rachael Clifford ◽  
Naren Govindarajah ◽  
David Bowden ◽  
Paul Sutton ◽  
Jason Parsons ◽  
...  
Keyword(s):  
Rectal Cancer ◽  
Locally Advanced ◽  
Advanced Rectal Cancer ◽  
Locally Advanced Rectal Cancer ◽  
Acid Ceramidase ◽  
Potential Biomarker

Acid ceramidase: a potential biomarker for locally advanced rectal cancer

2021 ◽  
Vol 108 (Supplement_5) ◽  
Author(s):  
Rachael Elizabeth Clifford ◽  
Naren Govindarajah ◽  
David Bowden ◽  
Paul Sutton ◽  
Jason Parsons ◽  
...  
Keyword(s):  
Rectal Cancer ◽  
Locally Advanced ◽  
Advanced Rectal Cancer ◽  
Locally Advanced Rectal Cancer ◽  
Acid Ceramidase ◽  
Potential Biomarker

scholarly journals LncRNAs Signature Associated with Chemoradiotherapy Response and Prognosis in Locally Advanced Rectal Cancer

2021 ◽  
Author(s):  
Yiyi Zhang ◽  
Binjie Guan ◽  
Yong Wu ◽  
Fan Du ◽  
Jinfu Zhuang ◽  
...  
Keyword(s):  
Rectal Cancer ◽  
Cell Lines ◽  
Expression Profiles ◽  
Locally Advanced ◽  
Advanced Rectal Cancer ◽  
Locally Advanced Rectal Cancer ◽  
Lncrna Expression ◽  
Cox Analysis

Abstract Background Long non-coding RNAs (lncRNAs) are promising diagnostic and prognostic biomarkers in cancers. Neoadjuvant chemoradiotherapy (NCRT) is the standard of care for patients with locally advanced rectal cancer (LARC). However, studies are limited regarding lncRNAs associated NCRT response and prognosis of LARC patients. This study aimed to identify lncRNAs associated with NCRT response and prognosis in CRC patients, and to explore potential mechanisms. Methods LncRNA expression profiles from our previous gene chip data basing on the LASSO to identify a four-lncRNA signature that predicted NCRT response and prognosis and further validated in 138 colorectal cancer (CRC) patients and 36 LARC patients from our center. A Cox regression model was performed to identify prognostic risk factors. Moreover, we identified the function of the LINC00909 in vivo and in vitro in CRC cell lines. Results Four hub lncRNAs (DBET, LINC00909, FLJ33534, and HSD52) were screened by comparing the relative lncRNA expression of NCRT-responsive and non-responsive patients (AUC = 0.68, 0.73, 0.73, and 0.70, respectively, all p < 0.05). A competing endogenous RNA (ceRNA) network was constructed based on the four lncRNAs. Moreover, the four lncRNAs expression was identified by the external data in cancerous and adjacent non-cancerous tissues in CRC patients. The results demonstrated that the expression of the four lncRNAs was lower in the normal tissues than in the cancerous tissues (all p < 0.05), and the COX analysis demonstrated that the DBET, LINC00909 and FLJ33534 were assocaited with the DFS in CRC patients. The four lncRNAs were also identified in the LARC following NCRT patients, and the result demonstrated that LINC00909 and FLJ33534 had powerful ability to predict the NCRT response and prognosis (all p < 0.05). Basing on the multivariate COX analysis, we constructed a risk score and verified in the CRC and LARC patients in predicting NCRT response and prognosis. Moreover, The expression and prognosis of the DBET, LINC00909 and FLJ33534 in the CRC tissues were identified in the R2 platform and Oncomine database. Moreover, the over-expression LINC00909 cell lines demonstrated that over-expressed the LINC00909 increased the cell lines resistance to the 5-FU and radiotherapy in vivo and in vitro. Conclusion Our findings showed that DBET, LINC00909, and FLJ33534 could serve as novel biomarkers for prediction of NCRT response and prognosis in CRC patients. And LINC00909 could be a novel therapeutic targets in enhancing the NCRT response.

Loss of RSPO3 as a potential mechanism of greater invasiveness in prostate cancer.

2018 ◽  
Vol 36 (6_suppl) ◽  
pp. 333-333
Author(s):  
Aruz Mesci ◽  
Xiaoyong Huang ◽  
David Shin ◽  
Christianne Hoey ◽  
Jessica Ray ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Lines ◽  
Metastatic Disease ◽  
Radiation Resistance ◽  
Locally Advanced ◽  
Prostate Adenocarcinoma ◽  
Potential Mechanism ◽  
Normal Prostate ◽  
Potential Biomarker

333 Background: While prostate cancer can often manifest as an indolent disease, locally-advanced or metastatic disease can cause significant morbidity or mortality. Better understanding of molecular mechanisms leading to more aggressive presentations can lead to identification of prognostic or predictive biomarkers, as well as to possible therapeutic targets. Methods: We describe the generation of prostate cancer cell lines that have been selected through a course of hypofractionated radiotherapy. We show that R-spondin 3, a protein implicated previously as a marker of aggressive disease in colorectal and non-small cell lung cancer, is downregulated in these cell lines compared to parental cells. Then, we test the functional consequences of R-spondin 3 downregulation by siRNA-mediated interference using in vitro assays of radiation resistance, proliferation, and invasiveness . Experiments investigating any change in metastatic propensity following R-spondin 3 downregulation are forthcoming. We further investigate signaling pathways that may be implicated in the mechanism of R-spondin 3 action. Lastly, we use the Oncomine Platform to look for any changes in R-spondin 3 levels between normal prostate tissue and prostate adenocarcinoma. Results: Although siRNA-mediated loss of R-spondin 3 does not confer radiation resistance in vitro, it does result in increased invasiveness in Matrigel assays. Paradoxically, R-spondin 3 downregulation results in decreased proliferation. Interference with R-spondin 3 also results in decreased β-catenin, ERK, and Akt signaling, but increased JNK signaling. In support of our in vitro results, R-spondin 3 expression is lower in human prostate adenocarcinoma compared to normal prostate in several gene expression datasets. Conclusions: Our preliminary data suggest that R-spondin 3 loss may lead to increased disease aggression via altered cell signaling. R-spondin 3 downregulation may represent a potential biomarker to identify patients at greater risk of developing locally advanced or metastatic disease, or a potential mechanism to target in future therapeutic efforts.

scholarly journals Hsa_circ_0005273 facilitates breast cancer tumorigenesis by regulating YAP1-hippo signaling pathway

2021 ◽  
Vol 40 (1) ◽  
Author(s):  
Xuehui Wang ◽  
Changle Ji ◽  
Jiashu Hu ◽  
Xiaochong Deng ◽  
Wenfang Zheng ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Signaling Pathway ◽  
Cell Lines ◽  
Luciferase Reporter ◽  
Circular Rnas ◽  
Hippo Signaling ◽  
Hippo Signaling Pathway ◽  
Potential Biomarker

Abstract Background Circular RNAs (circRNAs), a novel class of endogenous RNAs, have shown to participate in the development of breast cancer (BC). Hsa_circ_0005273 is a circRNA generated from several exons of PTK2. However, the potential functional role of hsa_circ_0005273 in BC remains largely unknown. Here we aim to evaluate the role of hsa_circ_0005273 in BC. Methods The expression level of hsa_circ_0005273 and miR-200a-3p were examined by RT-qPCR in BC tissues and cell lines. The effect of knocking down hsa_circ_0005273 in BC cell lines were evaluated by examinations of cell proliferation, migration and cell cycle. In addition, xenografts experiment in nude mice were performed to evaluate the effect of hsa_circ_0005273 in BC. RNA immunoprecipitation assay, RNA probe pull-down assay, luciferase reporter assay and fluorescence in situ hybridization were conducted to confirm the relationship between hsa_circ_0005273, miR-200a-3p and YAP1. Results Hsa_circ_0005273 is over-expressed in BC tissues and cell lines, whereas miR-200a-3p expression is repressed. Depletion of hsa_circ_0005273 inhibited the progression of BC cells in vitro and in vivo, while overexpression of hsa_circ_0005273 exhibited the opposite effect. Importantly, hsa_circ_0005273 upregulated YAP1 expression and inactivated Hippo pathway via sponging miR-200a-3p to promote BC progression. Conclusions Hsa_circ_0005273 regulates the miR-200a-3p/YAP1 axis and inactivates Hippo signaling pathway to promote BC progression, which may become a potential biomarker and therapeutic target.

scholarly journals EWS-FLI1-mediated tenascin-C expression promotes tumour progression by targeting MALAT1 through integrin α5β1-mediated YAP activation in Ewing sarcoma

2019 ◽  
Vol 121 (11) ◽  
pp. 922-933 ◽  
Author(s):  
Shaohui He ◽  
Quan Huang ◽  
Jinbo Hu ◽  
Lei Li ◽  
Yanbin Xiao ◽  
...  
Keyword(s):  
Cell Lines ◽  
Ewing Sarcoma ◽  
Nuclear Translocation ◽  
Tumour Progression ◽  
Underlying Mechanism ◽  
Integrin Α5β1 ◽  
Tenascin C ◽  
Tumorigenesis And Progression

Abstract Background The extracellular matrix has been critically associated with the tumorigenesis and progression of Ewing sarcoma (ES). However, the regulatory and prognostic roles of tenascin-C (TNC) in ES remain unclear. Methods TNC expression was examined in specimens by immunohistochemistry, and the association of TNC expression with ES patient survival was also analysed. TNC-knockout cell lines were constructed using CRISPR/Cas9 methods. In vitro experiments and in vivo bioluminescent imaging using BALB/c nude mice were conducted to evaluate the effect of TNC on ES tumour progression. RNA sequencing was performed, and the underlying mechanism of TNC was further explored. Results TNC was overexpressed in ES tissue and cell lines, and TNC overexpression was associated with poor survival in ES patients. TNC enhanced cell proliferation, migration and angiogenesis in vitro and promoted ES metastasis in vivo. The oncoprotein EWS-FLI1 profoundly increased TNC expression by directly binding to the TNC promoter region. Metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) upregulation induced by Yes-associated protein (YAP) activation was responsible for TNC-regulated ES tumour progression. Activated integrin α5β1 signalling might be correlated with YAP dephosphorylation and nuclear translocation. Conclusions TNC may promote ES tumour progression by targeting MALAT1 through integrin α5β1-mediated YAP activation.

scholarly journals Targeting Acid Ceramidase to Improve the Radiosensitivity of Rectal Cancer

Cells ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 2693
Author(s):  
Rachael E. Clifford ◽  
Naren Govindarajah ◽  
David Bowden ◽  
Paul Sutton ◽  
Mark Glenn ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Rectal Cancer ◽  
Cell Lines ◽  
Cancer Cell ◽  
Locally Advanced ◽  
Advanced Rectal Cancer ◽  
Cancer Cell Lines ◽  
Locally Advanced Rectal Cancer ◽  
Colorectal Cancer Cell ◽  
Colorectal Cancer Cell Lines

Previous work utilizing proteomic and immunohistochemical analyses has identified that high levels of acid ceramidase (AC) expression confers a poorer response to neoadjuvant treatment in locally advanced rectal cancer. We aimed to assess the radiosensitising effect of biological and pharmacological manipulation of AC and elucidate the underlying mechanism. AC manipulation in three colorectal cancer cell lines (HT29, HCT116 and LIM1215) was achieved using siRNA and plasmid overexpression. Carmofur and a novel small molecular inhibitor (LCL521) were used as pharmacological AC inhibitors. Using clonogenic assays, we demonstrate that an siRNA knockdown of AC enhanced X-ray radiosensitivity across all colorectal cancer cell lines compared to a non-targeting control siRNA, and conversely, AC protein overexpression increased radioresistance. Using CRISPR gene editing, we also generated AC knockout HCT116 cells that were significantly more radiosensitive compared to AC-expressing cells. Similarly, two patient-derived organoid models containing relatively low AC expression were found to be comparatively more radiosensitive than three other models containing higher levels of AC. Additionally, AC inhibition using carmofur and LCL521 in three colorectal cancer cell lines increased cellular radiosensitivity. Decreased AC protein led to significant poly-ADP ribose polymerase-1 (PARP-1) cleavage and apoptosis post-irradiation, which was shown to be executed through a p53-dependent process. Our study demonstrates that expression of AC within colorectal cancer cell lines modulates the cellular response to radiation, and particularly that AC inhibition leads to significantly enhanced radiosensitivity through an elevation in apoptosis. This work further solidifies AC as a target for improving radiotherapy treatment of locally advanced rectal cancer.

iMYC: Proof-of-concept study of ibrutinib in c-MYC and HER2 amplified oesophagogastric carcinoma.

2017 ◽  
Vol 35 (4_suppl) ◽  
pp. TPS221-TPS221
Author(s):  
Michael Davidson ◽  
Irene Yu-Shing Chong ◽  
David Cunningham ◽  
Lauren Aronson ◽  
Hanna Bryant ◽  
...  
Keyword(s):  
Cell Lines ◽  
Human Cancer ◽  
Locally Advanced ◽  
Imaging Biomarker ◽  
Cancer Drug ◽  
Lethal Gene ◽  
Her2 Amplification ◽  
Open Label ◽  
Biomarker Analysis

TPS221 Background: The MYC proto-oncogene is among the most frequently dysregulated genes in human cancer and is amplified in 10-30% of oesophagogastric (OG) cancers. Such genes which code for transcription factors are challenging to target directly with small molecule inhibitors or monoclonal antibodies. High-throughput siRNA screening of OG cell lines has found silencing of Bruton’s tyrosine kinase (BTK) to result in selective lethality in the presence of MYC amplification. Sensitivity to the orally available BTK inhibitor ibrutinib has been confirmed in-vitro in MYC, HER2 and co-amplified OG cancer cell lines (Chong, Ann Onc 2014), suggesting it may be a potentially effective therapeutic strategy in both MYC and HER2 amplified OG cancer. Methods: i MYC is an open label, phase II non-randomised study to assess the efficacy of ibrutinib in advanced pre-treated OG cancer. A novel FISH assay for tumour MYC amplification has been developed and pts are pre-screened for MYC and HER2 amplification. Eligibility includes metastatic or locally advanced inoperable OG cancer (SCC or adeno), MYC (ratio >2.5) and/or HER2 (ratio>2) amplification, progression after at least 1 prior line of chemotherapy +/- trastuzumab for advanced disease, PS 0-2 and no history of significant cardiovascular or bleeding disorders. Of the first 9 pts, at least 4 will demonstrate MYC amplification and remaining 5 will show either MYC or HER2 amplification, or co-amplification of both. Pts will be treated with ibrutinib monotherapy until progression or unacceptable toxicity. A Simon 2 stage design will be used for the primary endpoint of response rate, with interim analysis after 9 pts and maximum recruitment to 17 pts. Secondary endpoints include PFS, OS and exploratory translational and imaging biomarker analysis. Mandatory biopsy will be obtained at baseline and optional biopsies will be undertaken at day 10-14, week 8 and progression. The trial represents the first attempt at targeting MYC amplification via synthetic lethal gene interactions in OG cancer through the novel application of an existing anti-cancer drug. As of September 2016, 61 patients have consented for pre-screening, 19 completed MYC analysis and first patient has started treatment. Clinical trial information: 02884453.

NSABP FR-2: Phase II study of durvalumab following neoadjuvant chemoRT in stage II-IV rectal cancer.

2019 ◽  
Vol 37 (4_suppl) ◽  
pp. TPS727-TPS727
Author(s):  
Thomas J. George ◽  
Greg Yothers ◽  
James J. Lee ◽  
Samuel A. Jacobs ◽  
Melvin Deutsch ◽  
...  
Keyword(s):  
Rectal Cancer ◽  
Phase Ii ◽  
Phase Ii Trial ◽  
Locally Advanced ◽  
Neoadjuvant Chemoradiotherapy ◽  
Stage Iv ◽  
Sphincter Preservation ◽  
Stage Ii ◽  
Systemic Steroid ◽  
Clinical Role

TPS727 Background: Locally advanced rectal cancer remains a clinical challenge with few improvements noted over the past few decades. Although immunotherapy has no current clinical role in microsatellite stable (MSS) colorectal cancer, preclinical models suggest that radiotherapy (RT) can enhance neoantigen presentation, modulate the microenvironment, and improve the likelihood of anti-tumor activity with checkpoint inhibitor use. This prospective phase II trial will test that hypothesis in addition to confirming safety of this approach using a “window-of-opportunity” study design with the anti-PD-L1 agent durvalumab (MEDI4736). Methods: This multi-center phase II trial is currently enrolling patients (pts) with rectal cancer who are undergoing standard NCCN guideline-compliant neoadjuvant chemoradiotherapy (CRT). Eligibility includes pts with MSS stage II-IV rectal cancer with adequate organ function and pre-treatment diagnostic tumor available for profiling who are undergoing CRT with intentions to proceed to surgical resection. Stage IV disease must be limited such that the primary pelvic tumor requires definitive management. Standard ineligibility criteria include active infections, systemic steroid use, or other conditions making immunotherapy use unsafe. Treatment includes durvalumab (750mg IV infusion once every 2 wks) for 4 total doses beginning within 3-7 days after CRT completion. Surgery must be within 8-12 wks of the final CRT dose. Primary endpoint is a demonstrated improvement in Neoadjuvant Rectal Cancer (NAR) score compared to historical controls targeting a 20% relative risk reduction in DFS and 3-4% absolute OS improvement. Secondary endpoints include OS, DFS, toxicity, pCR, cCR, therapy completion, negative surgical margins, sphincter preservation, off-target “abscopal” effects for the subset of stage IV pts, and exploratory assessments of tumor infiltrating lymphocytes, circulating immunologic profiles, and molecular predictors of response. A safety run-in phase has completed as a precedent to full enrollment. Enrollment now continues to 47 total pts to achieve 41 surgically evaluable pts. NCT03102047. Support: AstraZeneca-Medimmune, NSABP Foundation Clinical trial information: NCT03102047.

scholarly journals The novel circ_0028171/miR-218-5p/IKBKB axis promotes osteosarcoma cancer progression

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Feng Pan ◽  
Jun Zhang ◽  
Benseng Tang ◽  
Li Jing ◽  
Bing Qiu ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Progression ◽  
Bone Cells ◽  
Human Cancer ◽  
Underlying Mechanism ◽  
Circular Rna ◽  
Luciferase Reporter ◽  
Migration And Invasion

Abstract Background Recently, it has been demonstrated that circular RNA (circRNA) contributes to the production and progression in human cancer. However, the specific function and underlying mechanism of circ_0028171 in osteosarcoma (OS) still remain largely unclear and require to be investigated. Methods In our study, we confirmed differentially expressed circRNAs by microarray analysis in normal bone cells vs. OS cell lines. The expression of circ-0028171 in OS was measured by qRT-PCR. Nuclear-cytoplasmic fractionation was employed to identify the localization of circ-0028171, and RNase R and actinomycin D treatment were used to prove its circular characteristic. In vitro experiments, such as CCK-8 method, cell count, cell colony formation, transwell migration and invasion assays, and in vivo tumor models were adopted to evaluate the effect of circ_0028171. Further, luciferase reporter, RIP and RNA pull-down assays were conducted to confirm the binding sites of circ_0028171 with miR-218-5p. Results We found that circ_0028171 displayed a remarkably higher expression in both OS tissues and cell lines. Circ_0028171 mainly located in the cytoplasm as a stable cyclic transcript. Knockdown of circ_0028171 suppressed OS tumor growth in vitro and in vivo, while up-regulated circ_0028171 remarkably enhanced cell proliferation, migration and invasion abilities in OS. Several mechanistic experiments revealed that circ_0028171 served as a sponge of miR-218-5p to increase IKBKB expression. Conclusions our research reveals that circ_0028171 might promote the malignant behavior of OS tissues through miR-218-5p/IKBKB axis, which could be a potential novel marker for early diagnosis of OS.

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