scholarly journals UBR-Box Containing Protein, UBR5, is Over-Expressed in Human Lung Adenocarcinoma and is a Potential Therapeutic Target

2020 ◽  
Author(s):  
Kumar Saurabh ◽  
Parag P Shah ◽  
Mark Doll ◽  
Leah J Siskind ◽  
Levi J. Beverly
Keyword(s):  
Lung Cancer ◽  
Lung Adenocarcinoma ◽  
Cell Lines ◽  
Nude Mice ◽  
Therapeutic Target ◽  
Human Lung ◽  
Human Patient ◽  
Adenocarcinoma Cell ◽  
Ubiquitination Pathway

Abstract Background:N-end rule ubiquitination pathway is known to be disrupted in many diseases, including cancer. UBR5, an E3 ubiquitin ligase, is mutated and/or overexpressed in human lung cancer cells suggesting its pathological role in cancer. Methods: We determined expression of UBR5 protein in multiple lung cancer cell lines and human patient samples.Using immunoprecipitation followed by mass spectrometry we determined the UBR5 interacting proteins. The impact of loss of UBR5 for lung adenocarcinoma cell lines was analyzed using cell viability, clonogenic assays and in vivo xenograft models in nude mice. Additional Western blot analysis was performed to assess the loss of UBR5 on downstream signaling. Statistical analysis was done by one-way ANOVA for in vitro studies and Wilcoxon paired t-test for tumor volumes in vivo.Results:We show variability of UBR5 expression levels in lung adenocarcinoma cell lines and in primary human patient samples.To gain better insight into the role that UBR5 may play in lung cancer progression we performed unbiased interactome analyses for UBR5. Dataindicate that UBR5 has a wide range of interacting protein partners that are known to be involved in critical cellular processes such as DNA damage, proliferation and cell cycle regulation. We have demonstrated thatshRNA-mediated lossof UBR5 decreases cell viabilityand clonogenic potential of lung adenocarcinoma cell lines.In addition, we founddecreased levels of activatedAKT signaling after the loss of UBR5 in lung adenocarcinoma cell lines using multiple means of UBR5 knockdown/knockout. Furthermore, we demonstrated that loss of UBR5 in lung adenocarcinoma cells results in significant reduction of tumor volume in nude mice. Conclusions:These findings demonstrate that deregulation of the N-end rule ubiquitination pathway plays a crucial role in the etiology of some human cancers, and blocking this pathway via UBR5-specific inhibitors, may represent a unique therapeutic target for human cancers.

scholarly journals UBR-Box containing protein, UBR5, is over-expressed in human lung adenocarcinoma and is a potential therapeutic target

2020 ◽  
Author(s):  
Kumar Saurabh ◽  
Parag P. Shah ◽  
Mark A. Doll ◽  
Leah J. Siskind ◽  
Levi J. Beverly
Keyword(s):  
Lung Cancer ◽  
Lung Adenocarcinoma ◽  
Cell Viability ◽  
Cell Lines ◽  
Therapeutic Target ◽  
Human Lung ◽  
Human Patient ◽  
Adenocarcinoma Cell ◽  
Ubiquitination Pathway

AbstractBackgroundN-end rule ubiquitination pathway is known to be disrupted in many diseases, including cancer. UBR5, an E3 ubiquitin ligase, is mutated and/or overexpressed in human lung cancer cells suggesting its pathological role in cancer.MethodsWe determined expression of UBR5 protein in multiple lung cancer cell lines and human patient samples. Using immunoprecipitation followed by mass spectrometry we determined the UBR5 interacting proteins. The impact of loss of UBR5 for lung adenocarcinoma cell lines was analyzed using cell viability, clonogenic assays and in vivo xenograft models in nude mice. Additional Western blot analysis was performed to assess the loss of UBR5 on downstream signaling. Statistical analysis was done by one-way ANOVA for in vitro studies and Wilcoxon paired t-test for tumor volumes in vivo.ResultsWe show variability of UBR5 expression levels in lung adenocarcinoma cell lines and in primary human patient samples. To gain better insight into the role that UBR5 may play in lung cancer progression we performed unbiased interactome analyses for UBR5. Data indicate that UBR5 has a wide range of interacting protein partners that are known to be involved in critical cellular processes such as DNA damage, proliferation and cell cycle regulation. We have demonstrated that shRNA-mediated loss of UBR5 decreases cell viability and clonogenic potential of lung adenocarcinoma cell lines. In addition, we found decreased levels of activated AKT signaling after the loss of UBR5 in lung adenocarcinoma cell lines using multiple means of UBR5 knockdown/knockout. Furthermore, we demonstrated that loss of UBR5 in lung adenocarcinoma cells results in significant reduction of tumor volume in nude mice.ConclusionsThese findings demonstrate that deregulation of the N-end rule ubiquitination pathway plays a crucial role in the etiology of some human cancers, and blocking this pathway via UBR5-specific inhibitors, may represent a unique therapeutic target for human cancers.

scholarly journals UBR-Box containing protein, UBR5, is over-expressed in human lung adenocarcinoma and is a potential therapeutic target

2020 ◽  
Author(s):  
Kumar Saurabh ◽  
Parag P Shah ◽  
Mark Doll ◽  
Leah J Siskind ◽  
Levi J. Beverly
Keyword(s):  
Lung Cancer ◽  
Lung Adenocarcinoma ◽  
Cell Viability ◽  
Cell Lines ◽  
Therapeutic Target ◽  
Human Lung ◽  
Human Patient ◽  
Adenocarcinoma Cell ◽  
Ubiquitination Pathway

Abstract Background: N-end rule ubiquitination pathway is known to be disrupted in many diseases, including cancer. UBR5, an E3 ubiquitin ligase, is mutated and/or overexpressed in human lung cancer cells suggesting its pathological role in cancer. Methods: We determined expression of UBR5 protein in multiple lung cancer cell lines and human patient samples. Using immunoprecipitation followed by mass spectrometry we determined the UBR5 interacting proteins. The impact of loss of UBR5 for lung adenocarcinoma cell lines was analyzed using cell viability, clonogenic assays and in vivo xenograft models in nude mice. Additional Western blot analysis was performed to assess the loss of UBR5 on downstream signaling. Statistical analysis was done by one-way ANOVA for in vitro studies and Wilcoxon paired t-test for tumor volumes in vivo. Results: We show variability of UBR5 expression levels in lung adenocarcinoma cell lines and in primary human patient samples. To gain better insight into the role that UBR5 may play in lung cancer progression we performed unbiased interactome analyses for UBR5. Data indicate that UBR5 has a wide range of interacting protein partners that are known to be involved in critical cellular processes such as DNA damage, proliferation and cell cycle regulation. We have demonstrated that shRNA-mediated loss of UBR5 decreases cell viability and clonogenic potential of lung adenocarcinoma cell lines. In addition, we found decreased levels of activated AKT signaling after the loss of UBR5 in lung adenocarcinoma cell lines using multiple means of UBR5 knockdown/knockout. Furthermore, we demonstrated that loss of UBR5 in lung adenocarcinoma cells results in significant reduction of tumor volume in nude mice. Conclusions: These findings demonstrate that deregulation of the N-end rule ubiquitination pathway plays a crucial role in the etiology of some human cancers, and blocking this pathway via UBR5-specific inhibitors, may represent a unique therapeutic target for human cancers.

scholarly journals UBR-Box containing protein, UBR5, is over-expressed in human lung adenocarcinoma and is a potential therapeutic target

2020 ◽  
Author(s):  
Kumar Saurabh ◽  
Parag P Shah ◽  
Mark Doll ◽  
Leah J Siskind ◽  
Levi J. Beverly
Keyword(s):  
Lung Cancer ◽  
Lung Adenocarcinoma ◽  
Cell Viability ◽  
Cell Lines ◽  
Therapeutic Target ◽  
Human Lung ◽  
Human Patient ◽  
Adenocarcinoma Cell ◽  
Ubiquitination Pathway

Abstract Background: N-end rule ubiquitination pathway is known to be disrupted in many diseases, including cancer. UBR5, an E3 ubiquitin ligase, is mutated and/or overexpressed in human lung cancer cells suggesting its pathological role in cancer. Methods: We determined expression of UBR5 protein in multiple lung cancer cell lines and human patient samples. Using immunoprecipitation followed by mass spectrometry we determined the UBR5 interacting proteins. The impact of loss of UBR5 for lung adenocarcinoma cell lines was analyzed using cell viability, clonogenic assays and in vivo xenograft models in nude mice. Additional Western blot analysis was performed to assess the loss of UBR5 on downstream signaling. Statistical analysis was done by one-way ANOVA for in vitro studies and Wilcoxon paired t-test for tumor volumes in vivo. Results: We show variability of UBR5 expression levels in lung adenocarcinoma cell lines and in primary human patient samples. To gain better insight into the role that UBR5 may play in lung cancer progression we performed unbiased interactome analyses for UBR5. Data indicate that UBR5 has a wide range of interacting protein partners that are known to be involved in critical cellular processes such as DNA damage, proliferation and cell cycle regulation. We have demonstrated that shRNA-mediated loss of UBR5 decreases cell viability and clonogenic potential of lung adenocarcinoma cell lines. In addition, we found decreased levels of activated AKT signaling after the loss of UBR5 in lung adenocarcinoma cell lines using multiple means of UBR5 knockdown/knockout. Furthermore, we demonstrated that loss of UBR5 in lung adenocarcinoma cells results in significant reduction of tumor volume in nude mice. Conclusions: These findings demonstrate that deregulation of the N-end rule ubiquitination pathway plays a crucial role in the etiology of some human cancers, and blocking this pathway via UBR5-specific inhibitors, may represent a unique therapeutic target for human cancers.

scholarly journals Tobacco carcinogen-induced mouse lung adenocarcinoma cell lines as tools to identify novel lung cancer genes

2018 ◽  
Author(s):  
Νικόλαος Κανελλάκης
Keyword(s):  
Lung Cancer ◽  
Lung Adenocarcinoma ◽  
Cell Lines ◽  
Mouse Lung ◽  
Cancer Genes ◽  
Adenocarcinoma Cell ◽  
Tobacco Carcinogen

Η νόσος του καρκίνου αποτελεί ένα σημαντικό και βαρύ κοινωνικό-οικονομικό φορτίο τόσο για τις οικονομικά αναπτυγμένες όσο και για τις οικονομικά αναπτυσσόμενες κοινωνίες. Η συχνότητα της διάγνωσης καρκινικών περιστατικών δείχνει να αυξάνεται με την πάροδο του χρόνου λόγω της γήρανσης του πληθυσμού και της αδυναμίας περιορισμού της χρήσης παραγόντων οι οποίοι έχουν συσχετιστεί με τον κίνδυνο εμφάνισης καρκίνου, όπως το κάπνισμα. Ο καρκίνος του πνεύμονα αποτελεί την κύρια αιτία θανάτου από καρκίνο παγκοσμίως. Οι κακοήθειες οι οποίες επάγονται από καρκινογόνα, συμπεριλαμβανομένου του πνευμονικού αδενοκαρκινώματος των καπνιστών, περιέχουν χιλιάδες μεταλλάξεις στο γονιδίωμά τους. Τις τελευταίες δεκαετίες έχουν ανακαλυφθεί και μελετηθεί πολλά νέα ογκογονίδια συσχετιζόμενα με το αδενοκαρκίνωμα πνεύμονα με αποτέλεσμα να εισαχθούν στην κλινική πράξη νέοι αντικαρκινικοί παράγοντες οι οποίοι άλλαξαν την πρόγνωση των ασθενών και βελτίωσαν την ποιότητα της ζωής τους. Ωστόσο, λόγω περιορισμών σχεδόν στα μισά περιστατικά τα ογκογονίδια τα οποία ευθύνονται για την ανάπτυξη των νεοπλασμάτων δεν είναι δυνατό να εντοπιστούν και έτσι παραμένουν άγνωστα. Παρά την ύπαρξη περίτεχνων γενετικών μοντέλων ποντικών του καρκίνου του πνεύμονα, αδυνατούν να προσομοιώσουν την ανθρώπινη νόσο με ακρίβεια και πιστότητα. Η ανάπτυξη ενός αξιόπιστου μοντέλου ποντικών το οποίο θα μπορούσε να χρησιμοποιηθεί για την εντοπισμό των ογκογονιδίων και μεταλλάξεων του καρκίνου πνεύμονα παραμένει μια μη ικανοποιημένη και ιδιαίτερα σημαντική ερευνητική ανάγκη. Καταφέραμε να αναπτύξουμε μια μεθοδολογία για τη δημιουργία κυτταρικών σειρών αδενοκαρκινώματος πνεύμονα ποντικού από διάφορα στελέχη μετά από επαναλαμβανόμενη έκθεση στις καρκινογόνες ουσίες του καπνού: ουρεθάνη και διαιθυλο νιτροζαμίνη. Οι κυτταρικές σειρές οι οποίες προέκυψαν (n = 7) ήταν αθάνατες και φαινοτυπικά σταθερές in vitro, ογκογόνες, μεταστατικές και θανατηφόρες in vivo. Οι επαγόμενες από χημικά καρκινογόνα του καπνού κυτταρικές σειρές παρουσίασαν αξιοσημείωτη ομοιότητα με το ανθρώπινο αδενοκαρκίνωμα πνεύμονα, καθώς περιείχαν μεταλλάξεις στα γονίδια Kras και Trp53 και παρουσίασαν χαρακτηριστικές ιδιότητες των καρκινικών βλαστοκυττάρων. Ανακαλύψαμε ότι όλες οι κυτταρικές σειρές υπερέκφραζαν το γονίδιο Proliferin, ένα ορθολογo στον ποντικό για την ανθρώπινη προλακτίνη. Η λειτουργική μελέτη του γονιδίου έδειξε πως δρα ως υποκινητής για την αναπτυξη πνευμονικών όγκων. Είναι ενδιαφέρον το γεγονός πως η προλακτίνη βρέθηκε υπερεκφρασμένη και συσχετίστηκε με κακή πρόγνωση στο ανθρώπινο αδενοκαρκίνωμα πνεύμονα. Συμπερασματικά τα κύτταρα αδενοκαρκινώματος πνεύμονα ποντικού τα οποία δημιουργήσαμε με τη χρήση καρκινογόνων του καπνού μοιάζουν με την ανθρώπινη νόσο του αδενοκαρκινώματος πνεύμονα. Ως εκ τούτου μπορούν να χρησιμοποιηθούν ως ένα πολύτιμο εργαλείο για την ανακάλυψη και τη λειτουργική διερεύνηση νέων ογκογονιδίων του αδενοκαρκινώματος του πνεύμονα, των μεταλλάξεων που οδηγούν στην ανάπτυξή του και των βιολογικών μονοπατιών και μηχανισμών που προάγουν την πνευμονική καρκινογένεση.ΛΕΞΕΙΣ ΚΛΕΙΔΙΑπνεύμονας; αδενοκαρκίνωμα; καπνός; καρκινογόνο; κάπνισμα; KRAS; TRP53

scholarly journals Cisplatin loaded multiwalled carbon nanotubes reverse drug resistance in NSCLC by inhibiting EMT

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Yuxin Qi ◽  
Wenping Yang ◽  
Shuang Liu ◽  
Fanjie Han ◽  
Haibin Wang ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Carbon Nanotubes ◽  
Drug Resistance ◽  
Western Blot ◽  
Multiwalled Carbon Nanotubes ◽  
Cell Lines ◽  
Nude Mice ◽  
Expression Level ◽  
Multiwalled Carbon

Abstract Background Lung cancer is one of the important health threats worldwide, of which 5-year survival rate is less than 15%. Non-small-cell lung cancer (NSCLC) accounts for about 80% of all lung cancer with high metastasis and mortality. Methods Cisplatin loaded multiwalled carbon nanotubes (Pt-MWNTS) were synthesized and used to evaluate the anticancer effect in our study. The NSCLC cell lines A549 (cisplatin sensitive) and A549/DDP (cisplatin resistant) were used in our in vitro assays. MTT was used to determine Cancer cells viability and invasion were measured by MTT assay and Transwell assay, respectively. Apoptosis and epithelial-mesenchymal transition related marker proteins were measured by western blot. The in vivo anti-cancer effect of Pt-MWNTs were performed in male BALB/c nude mice (4-week old). Results Pt-MWNTS were synthesized and characterized by X-ray diffraction, Raman, FT-IR spectroscopy and scan electron microscopy. No significant cytotoxicity of MWNTS was detected in both A549/DDP and A549 cell lines. However, Pt-MWNTS showed a stronger inhibition effect on cell growth than free cisplatin, especially on A549/DDP. We found Pt-MWNTS showed higher intracellular accumulation of cisplatin in A549/DDP cells than free cisplatin and resulted in enhanced the percent of apoptotic cells. Western blot showed that application of Pt-MWNTS can significantly upregulate the expression level of Bax, Bim, Bid, Caspase-3 and Caspase-9 while downregulate the expression level of Bcl-2, compared with free cisplatin. Moreover, the expression level of mesenchymal markers like Vimentin and N-cadherin was more efficiently reduced by Pt-MWNTS treatment in A549/DDP cells than free cisplatin. In vivo study in nude mice proved that Pt-MWNTS more effectively inhibited tumorigenesis compared with cisplatin, although both of them had no significant effect on body weight. Conclusion Pt-MWNT reverses the drug resistance in the A549/DDP cell line, underlying its possibility of treating NSCLC with cisplatin resistance.

scholarly journals The Anti-Cancer Effects of a Zotarolimus and 5-Fluorouracil Combination Treatment on A549 Cell-Derived Tumors in BALB/c Nude Mice

2021 ◽  
Vol 22 (9) ◽  
pp. 4562
Author(s):  
Ching-Feng Wu ◽  
Ching-Yang Wu ◽  
Robin Y.-Y. Chiou ◽  
Wei-Cheng Yang ◽  
Chuen-Fu Lin ◽  
...  
Keyword(s):  
Growth Factor ◽  
Tumor Growth ◽  
Lung Adenocarcinoma ◽  
Nude Mice ◽  
Transforming Growth Factor ◽  
Human Lung ◽  
Transforming Growth Factor Β ◽  
Related Factors ◽  
Human Lung Adenocarcinoma

Zotarolimus is a semi-synthetic derivative of rapamycin and a novel immunosuppressive agent used to prevent graft rejection. The pharmacological pathway of zotarolimus restricts the kinase activity of the mammalian target of rapamycin (mTOR), which potentially leads to reductions in cell division, cell growth, cell proliferation, and inflammation. These pathways have a critical influence on tumorigenesis. This study aims to examine the anti-tumor effect of zotarolimus or zotarolimus combined with 5-fluorouracil (5-FU) on A549 human lung adenocarcinoma cell line implanted in BALB/c nude mice by estimating tumor growth, apoptosis expression, inflammation, and metastasis. We established A549 xenografts in nude mice, following which we randomly divided the mice into four groups: control, 5-FU (100 mg/kg/week), zotarolimus (2 mg/kg/day), and zotarolimus combined with 5-FU. Compared the results with those for control mice, we found that mice treated with zotarolimus or zotarolimus combined with 5-FU retarded tumor growth; increased tumor apoptosis through the enhanced expression of cleaved caspase 3 and extracellular signal-regulated kinase (ERK) phosphorylation; decreased inflammation cytokines levels (e.g., IL-1β, TNF-α, and IL-6); reduced inflammation-related factors such as cyclooxygenase-2 (COX-2) protein and nuclear factor-κB (NF-κB) mRNA; enhanced anti-inflammation-related factors including IL-10 and inhibitor of NF-κB kinase α (IκBα) mRNA; and inhibited metastasis-related factors such as transforming growth factor β (TGF-β), CD44, epidermal growth factor receptor (EGFR), and vascular endothelial growth factor (VEGF). Notably, mice treated with zotarolimus combined with 5-FU had significantly retarded tumor growth, reduced tumor size, and increased tumor inhibition compared with the groups of mice treated with 5-FU or zotarolimus alone. The in vivo study confirmed that zotarolimus or zotarolimus combined with 5-FU could retard lung adenocarcinoma growth and inhibit tumorigenesis. Zotarolimus and 5-FU were found to have an obvious synergistic tumor-inhibiting effect on lung adenocarcinoma. Therefore, both zotarolimus alone and zotarolimus combined with 5-FU may be potential anti-tumor agents for treatment of human lung adenocarcinoma.

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