المؤلفون: Upasana Ray, Debarshi Roy, Ling Jin, Prabhu Thirusangu, Julie Staub, Yinan Xiao, Eleftheria Kalogera, Andrea E. Wahner Hendrickson, Grace D. Cullen, Krista Goergen, Ann L. Oberg, Viji Shridhar

المصدر: Journal of Experimental & Clinical Cancer Research, Vol 40, Iss 1, Pp 1-16 (2021)

مصطلحات موضوعية: Ovarian cancer, metastasis, Group III phospholipase A2, chemosensitivity, autophagy, primary cilia, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282

الوصف: Abstract Background Aberrant lipogenicity and deregulated autophagy are common in most advanced human cancer and therapeutic strategies to exploit these pathways are currently under consideration. Group III Phospholipase A2 (sPLA2-III/PLA2G3), an atypical secretory PLA2, is recognized as a regulator of lipid metabolism associated with oncogenesis. Though recent studies reveal that high PLA2G3 expression significantly correlates with poor prognosis in several cancers, however, role of PLA2G3 in ovarian cancer (OC) pathogenesis is still undetermined. Methods CRISPR-Cas9 and shRNA mediated knockout and knockdown of PLA2G3 in OC cells were used to evaluate lipid droplet (LD) biogenesis by confocal and Transmission electron microscopy analysis, and the cell viability and sensitization of the cells to platinum-mediated cytotoxicity by MTT assay. Regulation of primary ciliation by PLA2G3 downregulation both genetically and by metabolic inhibitor PFK-158 induced autophagy was assessed by immunofluorescence-based confocal analysis and immunoblot. Transient transfection with GFP-RFP-LC3B and confocal analysis was used to assess the autophagic flux in OC cells. PLA2G3 knockout OVCAR5 xenograft in combination with carboplatin on tumor growth and metastasis was assessed in vivo. Efficacy of PFK158 alone and with platinum drugs was determined in patient-derived primary ascites cultures expressing PLA2G3 by MTT assay and immunoblot analysis. Results Downregulation of PLA2G3 in OVCAR8 and 5 cells inhibited LD biogenesis, decreased growth and sensitized cells to platinum drug mediated cytotoxicity in vitro and in in vivo OVCAR5 xenograft. PLA2G3 knockdown in HeyA8MDR-resistant cells showed sensitivity to carboplatin treatment. We found that both PFK158 inhibitor-mediated and genetic downregulation of PLA2G3 resulted in increased number of percent ciliated cells and inhibited cancer progression. Mechanistically, we found that PFK158-induced autophagy targeted PLA2G3 to restore primary cilia in OC cells. Of clinical relevance, PFK158 also induces percent ciliated cells in human-derived primary ascites cells and reduces cell viability with sensitization to chemotherapy. Conclusions Taken together, our study for the first time emphasizes the role of PLA2G3 in regulating the OC metastasis. This study further suggests the therapeutic potential of targeting phospholipases and/or restoration of PC for future OC treatment and the critical role of PLA2G3 in regulating ciliary function by coordinating interface between lipogenesis and metastasis.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/1756-9966Test

الوصول الحر: https://doaj.org/article/c51f205913f048ecae91afeecbaec90fTest

المؤلفون: Prabhu Thirusangu, Christopher L. Pathoulas, Upasana Ray, Yinan Xiao, Julie Staub, Ling Jin, Ashwani Khurana, Viji Shridhar

المصدر: Cancers, Vol 13, Iss 9, p 2004 (2021)

مصطلحات موضوعية: quinacrine, autophagy, CTSL, LMP, MOMP, ovarian cancer, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282

الوصف: We previously reported that the antimalarial compound quinacrine (QC) induces autophagy in ovarian cancer cells. In the current study, we uncovered that QC significantly upregulates cathepsin L (CTSL) but not cathepsin B and D levels, implicating the specific role of CTSL in promoting QC-induced autophagic flux and apoptotic cell death in OC cells. Using a Magic Red® cathepsin L activity assay and LysoTracker red, we discerned that QC-induced CTSL activation promotes lysosomal membrane permeability (LMP) resulting in the release of active CTSL into the cytosol to promote apoptotic cell death. We found that QC-induced LMP and CTSL activation promotes Bid cleavage, mitochondrial outer membrane permeabilization (MOMP), and mitochondrial cytochrome-c release. Genetic (shRNA) and pharmacological (Z-FY(tBU)-DMK) inhibition of CTSL markedly reduces QC-induced autophagy, LMP, MOMP, apoptosis, and cell death; whereas induced overexpression of CTSL in ovarian cancer cell lines has an opposite effect. Using recombinant CTSL, we identified p62/SQSTM1 as a novel substrate of CTSL, suggesting that CTSL promotes QC-induced autophagic flux. CTSL activation is specific to QC-induced autophagy since no CTSL activation is seen in ATG5 knockout cells or with the anti-malarial autophagy-inhibiting drug chloroquine. Importantly, we showed that upregulation of CTSL in QC-treated HeyA8MDR xenografts corresponds with attenuation of p62, upregulation of LC3BII, cytochrome-c, tBid, cleaved PARP, and caspase3. Taken together, the data suggest that QC-induced autophagy and CTSL upregulation promote a positive feedback loop leading to excessive autophagic flux, LMP, and MOMP to promote QC-induced cell death in ovarian cancer cells.

وصف الملف: electronic resource

العلاقة: https://www.mdpi.com/2072-6694/13/9/2004Test; https://doaj.org/toc/2072-6694Test

الوصول الحر: https://doaj.org/article/29e385f4fc1f41ab8ac36862db2e3b8cTest

المؤلفون: Upasana Ray (3763864), Debarshi Roy (10920043), Ling Jin (133762), Prabhu Thirusangu (10920046), Julie Staub (10920049), Yinan Xiao (7336949), Eleftheria Kalogera (10920052), Andrea E. Wahner Hendrickson (10920055), Grace D. Cullen (10920058), Krista Goergen (277812), Ann L. Oberg (217897), Viji Shridhar (278465)

مصطلحات موضوعية: Biophysics, Biochemistry, Microbiology, Cell Biology, Genetics, Molecular Biology, Physiology, Pharmacology, Biotechnology, Cancer, Hematology, Virology, Biological Sciences not elsewhere classified, Chemical Sciences not elsewhere classified, Ovarian cancer, metastasis, Group III phospholipase A2, chemosensitivity, autophagy, primary cilia

الوصف: Additional file 1.

العلاقة: https://figshare.com/articles/journal_contribution/Additional_file_1_of_Group_III_phospholipase_A2_downregulation_attenuated_survival_and_metastasis_in_ovarian_cancer_and_promotes_chemo-sensitization/14728585Test

الإتاحة: https://doi.org/10.6084/m9.figshare.14728585.v1Test

المؤلفون: Prabhu Thirusangu, Upasana Ray, Sayantani Sarkar Bhattacharya, Derek B. Oien, Ling Jin, Julie Staub, Nagarajan Kannan, Julian R. Molina, Viji Shridhar

المصدر: Oncogene. 42:79-82

مصطلحات موضوعية: Cancer Research, Genetics, Molecular Biology

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::6aac989b1b1b826b70fbe03fbb2811b6Test
https://doi.org/10.1038/s41388-022-02470-zTest

المؤلفون: Sayantani Sarkar Bhattacharya, Prabhu Thirusangu, Ling Jin, Julie Staub, Viji Shridhar, Julian R. Molina

المصدر: Br J Cancer

مصطلحات موضوعية: STAT3 Transcription Factor, Cancer Research, Oncology, Phosphofructokinase-2, Pyridines, Cell Line, Tumor, SOXB1 Transcription Factors, Mesothelioma, Malignant, Neoplastic Stem Cells, Quinolines, Humans, Article, Cell Proliferation

الوصف: BACKGROUND: Malignant pleural mesothelioma (MPM) is an aggressive neoplasm and often acquires chemoresistance by increasing stemness in tumour tissue, thereby generating cancer stem cells (CSCs). CSCs escape treatment by deploying metabolic pathways to trigger dormancy or proliferation, also gaining the ability to exit and re-enter the cell cycle to hide their cellular identity. METHODS: We employed various cellular and biochemical assays to identify the role of the glycolytic enzyme PFKFB3, by knocking it down and pharmacologically inhibiting it with PFK158, to determine its anticancer effects in vitro and in vivo by targeting the CSC population in MPM. RESULTS: Here, we have identified PFKFB3 as a strategic player to target the CSC population in MPM and demonstrated that both pharmacologic (PFK158) and genetic inhibition of PFKFB3 destroy the FAK-Stat3-SOX2 nexus resulting in a decline in conspicuous stem cell markers viz. ALDH, CD133, CD44, SOX2. Inhibition of PFKFB3 accumulates p21 and p27 in the nucleus by decreasing SKP2. Lastly, PFK158 diminishes tumour-initiating cells (TICs) mediated MPM xenograft in vivo. CONCLUSIONS: This study confers a comprehensive and mechanistic function of PFKFB3 in CSC maintenance that may foster exceptional opportunities for targeted small molecule blockade of the TICs in MPM.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a628e7e25775b1c3fccf109e312f073dTest
https://doi.org/10.1038/s41416-022-01867-7Test

المؤلفون: Viji Shridhar, Julian Molina, Julie Staub, Jacie Maguire, Alfonso Baldi, Mariarosaria Boccellino, Lucio Quagliuolo, Ramandeep Rattan, Zhixue Liu, Daniah Beleford

الوصف: Supplementary Data from Methylation Induced Gene Silencing of HtrA3 in Smoking-Related Lung Cancer

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5c18d425098a3aef07be40bd7405ae46Test
https://doi.org/10.1158/1078-0432.22440148.v1Test

المؤلفون: Viji Shridhar, Julian Molina, Julie Staub, Jacie Maguire, Alfonso Baldi, Mariarosaria Boccellino, Lucio Quagliuolo, Ramandeep Rattan, Zhixue Liu, Daniah Beleford

الوصف: Purpose: Some 85% of lung cancers are smoking related. Here, we investigate the role of serine protease HtrA3 in smoking-related lung cancer.Experimental Design: We assess HtrA3 methylation and its corresponding expression in the human bronchial cell line BEAS-2B following cigarette smoke carcinogen treatment, in lung cancer cell lines and in primary lung tumors from light, moderate, and heavy smokers. We also show the effects of HtrA3 downregulation on MTT reduction and clonogenic survival with etoposide and cisplatin treatment and the corresponding effects of HtrA3 re-expression during treatment.Results: We show for the first time that HtrA3 expression is reduced or completely lost in over 50% of lung cancer cell lines and primary lung tumors from heavy smokers. Treatment of HtrA3-deficient cell lines with 5-aza-2′-deoxycytidine resulted in a dose-dependent increase in HtrA3 transcription. Further, sequence analysis of bisulfite-modified DNA from lung cancer cell lines and from primary lung tumors showed an increased frequency of methylation within the first exon of HtrA3 with a corresponding loss of HtrA3 expression, particularly in tumors from smokers. In BEAS-2B, treatment with the cigarette smoke carcinogen 4-(methylnitrosamino)-I-(3-pyridyl)-1-butanone resulted in HtrA3 downregulation with a corresponding increase in methylation. Additional studies indicate resistance to etoposide and cisplatin cytotoxicity as a functional consequence of HtrA3 loss. Finally, immunohistochemical analysis of primary lung tumors revealed a strong correlation between low HtrA3 expression and heavy smoking history.Conclusions: Collectively, these results suggest that cigarette smoke–induced methylation of HtrA3 could contribute to the etiology of chemoresistant disease in smoking-related lung cancer. Clin Cancer Res; 16(2); 398–409

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::72b405d35971ed7a1300f02fd00f8a17Test
https://doi.org/10.1158/1078-0432.c.6517516.v1Test

المؤلفون: Jessica Weng, John Kisiel, Seth Slettedahl, Douglas Mahoney, Xiaoming Cao, Patrick Foote, Kelli Burger, Calise Berger, Maureen Lemens, Vijayalakshmi Shridhar, Julie Staub, Maria O’Connell, Karen Doering, J Kenneth Schoolmeester, Sarah Kerr, Rondell Graham, William Taylor, Mark Sherman, Jamie Bakkum-Gamez

المصدر: Featured Printed Posters (Poster Rounds with the Professors).

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::f3e61a7a20b8905afff43e74f1980b8fTest
https://doi.org/10.1136/ijgc-2022-igcs.86Test

المؤلفون: Debarshi Roy, Prabhu Thirusangu, Ann L. Oberg, Krista M. Goergen, Viji Shridhar, Andrea E. Wahner Hendrickson, Grace D. Cullen, Upasana Ray, Yinan Xiao, Ling Jin, Julie Staub, Eleftheria Kalogera

المصدر: Journal of Experimental & Clinical Cancer Research, Vol 40, Iss 1, Pp 1-16 (2021)
Journal of Experimental & Clinical Cancer Research : CR

مصطلحات موضوعية: Cancer Research, autophagy, Cell Survival, Pyridines, medicine.disease_cause, Metastasis, Mice, primary cilia, Downregulation and upregulation, Microscopy, Electron, Transmission, Ovarian cancer, medicine, Animals, Humans, metastasis, MTT assay, Viability assay, Neoplasm Metastasis, Cytotoxicity, RC254-282, Cell Proliferation, Platinum, Ovarian Neoplasms, Chemistry, Research, Group III Phospholipases A2, Lipogenesis, Cancer, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, Lipid Droplets, medicine.disease, Gene Expression Regulation, Neoplastic, chemosensitivity, Group III phospholipase A2, Oncology, Cancer research, Quinolines, Heterografts, Female, CRISPR-Cas Systems, Carcinogenesis

الوصف: Background Aberrant lipogenicity and deregulated autophagy are common in most advanced human cancer and therapeutic strategies to exploit these pathways are currently under consideration. Group III Phospholipase A2 (sPLA2-III/PLA2G3), an atypical secretory PLA2, is recognized as a regulator of lipid metabolism associated with oncogenesis. Though recent studies reveal that high PLA2G3 expression significantly correlates with poor prognosis in several cancers, however, role of PLA2G3 in ovarian cancer (OC) pathogenesis is still undetermined. Methods CRISPR-Cas9 and shRNA mediated knockout and knockdown of PLA2G3 in OC cells were used to evaluate lipid droplet (LD) biogenesis by confocal and Transmission electron microscopy analysis, and the cell viability and sensitization of the cells to platinum-mediated cytotoxicity by MTT assay. Regulation of primary ciliation by PLA2G3 downregulation both genetically and by metabolic inhibitor PFK-158 induced autophagy was assessed by immunofluorescence-based confocal analysis and immunoblot. Transient transfection with GFP-RFP-LC3B and confocal analysis was used to assess the autophagic flux in OC cells. PLA2G3 knockout OVCAR5 xenograft in combination with carboplatin on tumor growth and metastasis was assessed in vivo. Efficacy of PFK158 alone and with platinum drugs was determined in patient-derived primary ascites cultures expressing PLA2G3 by MTT assay and immunoblot analysis. Results Downregulation of PLA2G3 in OVCAR8 and 5 cells inhibited LD biogenesis, decreased growth and sensitized cells to platinum drug mediated cytotoxicity in vitro and in in vivo OVCAR5 xenograft. PLA2G3 knockdown in HeyA8MDR-resistant cells showed sensitivity to carboplatin treatment. We found that both PFK158 inhibitor-mediated and genetic downregulation of PLA2G3 resulted in increased number of percent ciliated cells and inhibited cancer progression. Mechanistically, we found that PFK158-induced autophagy targeted PLA2G3 to restore primary cilia in OC cells. Of clinical relevance, PFK158 also induces percent ciliated cells in human-derived primary ascites cells and reduces cell viability with sensitization to chemotherapy. Conclusions Taken together, our study for the first time emphasizes the role of PLA2G3 in regulating the OC metastasis. This study further suggests the therapeutic potential of targeting phospholipases and/or restoration of PC for future OC treatment and the critical role of PLA2G3 in regulating ciliary function by coordinating interface between lipogenesis and metastasis.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d2312f39716829d872e4f2d1f69a840aTest
https://doaj.org/article/c51f205913f048ecae91afeecbaec90fTest

المؤلفون: Yinan Xiao, Viji Shridhar, Julie Staub, Chaolin Deng, Sayantani Sarkar Bhattacharya, Upasana Ray, Ling Jin, Xiaoling Fang, Eleftheria Kalogera, Haotian Xu, Ye Guan, Prabhu Thirusangu

المصدر: Oncogene

مصطلحات موضوعية: Cancer Research, Programmed cell death, Phosphofructokinase-2, Pyridines, Apoptosis, Biology, Article, Carboplatin, chemistry.chemical_compound, Targeted therapies, Endometrial cancer, Downregulation and upregulation, Cell Line, Tumor, Autophagy, Genetics, medicine, Chemotherapy, Humans, Molecular Biology, Protein kinase B, Cell Proliferation, Cisplatin, Cell growth, TOR Serine-Threonine Kinases, Xenograft Model Antitumor Assays, Endometrial Neoplasms, Gene Expression Regulation, Neoplastic, chemistry, Drug Resistance, Neoplasm, Cell culture, Quinolines, Cancer research, Female, Signal Transduction, medicine.drug

الوصف: The advanced or recurrent endometrial cancer (EC) has a poor prognosis because of chemoresistance. 6-Phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3), a glycolytic enzyme, is overexpressed in a variety of human cancers and plays important roles in promoting tumor cell growth. Here, we showed that high expression of PFKFB3 in EC cell lines is associated with chemoresistance. Pharmacological inhibition of PFKFB3 with PFK158 and or genetic downregulation of PFKFB3 dramatically suppressed cell proliferation and enhanced the sensitivity of EC cells to carboplatin (CBPt) and cisplatin (Cis). Moreover, PFKFB3 inhibition resulted in reduced glucose uptake, ATP production, and lactate release. Notably, we found that PFK158 with CBPt or Cis exerted strong synergistic antitumor activity in chemoresistant EC cell lines, HEC-1B and ARK-2 cells. We also found that the combination of PFK158 and CBPt/Cis induced apoptosis- and autophagy-mediated cell death through inhibition of the Akt/mTOR signaling pathway. Mechanistically, we found that PFK158 downregulated the CBPt/Cis-induced upregulation of RAD51 expression and enhanced CBPt/Cis-induced DNA damage as demonstrated by an increase in γ-H2AX levels in HEC-1B and ARK-2 cells, potentially revealing a means to enhance PFK158-induced chemosensitivity. More importantly, PFK158 treatment, either as monotherapy or in combination with CBPt, led to a marked reduction in tumor growth in two chemoresistant EC mouse xenograft models. These data suggest that PFKFB3 inhibition alone or in combination with standard chemotherapy may be used as a novel therapeutic strategy for improved therapeutic efficacy and outcomes of advanced and recurrent EC patients.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9889ad4045dd12b8f7e423ae5438c349Test
https://doi.org/10.1038/s41388-020-01621-4Test