scholarly journals GLUT-1 siRNA Enhances Radiosensitization Of Laryngeal Cancer Stem Cells Via Enhanced DNA Damage, Cell Cycle Redistribution, And Promotion Of Apoptosis In Vitro And In Vivo

2019 ◽  
Vol Volume 12 ◽  
pp. 9129-9142 ◽  
Author(s):  
Jiang-Tao Zhong ◽  
Qi Yu ◽  
Shui-Hong Zhou ◽  
Er Yu ◽  
Yang-Yang Bao ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Dna Damage ◽  
Cancer Stem Cells ◽  
Laryngeal Cancer ◽  
Damage Cell ◽  
Glut 1

scholarly journals STEM-20. RADIO-SENSITIZING GLIOBLASTOMA CANCER STEM CELLS BY INHIBITION OF FACT

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi237-vi238
Author(s):  
Miranda Montgomery ◽  
Abigail Zalenski ◽  
Amanda Deighen ◽  
Sherry Mortach ◽  
Treg Grubb ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dna Damage ◽  
Cancer Stem Cells ◽  
Combination Therapy ◽  
High Rate ◽  
Primary Role ◽  
Cancer Cell Growth ◽  
Treatment Paradigm

Abstract Glioblastoma (GBM) has a particularly high rate of recurrence with a 5-year overall survival rate of approximately 5%. This is in part due to a sub-population of cancer stem cells (CSC), which are both radioresistant and chemotherapeutically resistant to conventional treatments. Here we investigated CBL0137, a small molecule form of curaxin, in combination with radiotherapy as a means to radiosensitize CSCs. CBL0137 sequesters FACT (facilitates chromatin transcription) complex to chromatin, which leads to activation of p53 and inhibition of NF-κB. This sequestering of FACT results in cytotoxicity especially within tumor cells and prevents FACT from performing its primary role as a histone chaperone, as well as inhibits its part in the DNA damage response pathway. We show that when combined with radiotherapy, CBL0137 administration limited the ability of CSCs to identify and repair damaged DNA. CSCs treated in vitro with CBL0137 and irradiation showed an increased inhibition of cancer cell growth and decreased viability compared to irradiation or drug alone. Combination therapy also showed more DNA damage in the CSCs than with either agent alone. Based on our in vitro evidence for the efficacy of combination therapy to target CSCs, we moved forward to test the treatment in vivo. Using a subcutaneous model, we show that the amount of CD133+ cells (a marker for GMB CSCs) was reduced in irradiation plus CBL0137 compared to either treatment alone. Survival studies demonstrated that irradiation plus CBL0137 compared to irradiation alone or CBL0137 alone increase lifespan. Here we show the ability of CBL0137, in combination with irradiation, to target patient GBM CSCs both in vitro and in vivo. This work establishes a new treatment paradigm for GBM that inclusively targets CSCs and may ultimately reduce tumor recurrence.

scholarly journals ID: 1023 All-trans retinoic acid targets gastric cancer stem cells and inhibits patient-derived gastric carcinoma tumor growth

2017 ◽  
Vol 4 (S) ◽  
pp. 98
Author(s):  
P H Nguyen ◽  
J Giraud ◽  
C Staedel ◽  
L Chambonnier ◽  
P Dubus ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Cancer Stem Cells ◽  
Gastric Carcinoma ◽  
Tumor Growth ◽  
Cell Cycle Progression ◽  
3D Culture ◽  
All Trans Retinoic Acid

Gastric carcinoma is the third leading cause of cancer-related death worldwide. This cancer, most of the time metastatic, is essentially treated by surgery associated with conventional chemotherapy, and has a poor prognosis. The existence of cancer stem cells (CSC) expressing CD44 and a high aldehyde dehydrogenase (ALDH) activity has recently been demonstrated in gastric carcinoma and has opened new perspectives to develop targeted therapy. In this study, we evaluated the effects of all-transretinoic acid (ATRA) on CSCs in human gastric carcinoma. ATRA effects were evaluated on the proliferation and tumorigenic properties of gastric carcinoma cells from patient-derived tumors and cell lines in conventional 2D cultures, in 3D culture systems (tumorsphere assay) and in mouse xenograft models. ATRA inhibited both tumorspheres initiation and growth in vitro, which was associated with a cell-cycle arrest through the upregulation of cyclin-dependent kinase (CDK) inhibitors and the downregulation of cell-cycle progression activators. More importantly, ATRA downregulated the expression of the CSC markers CD44 and ALDH as well as stemness genes such as Klf4 and Sox2 and induced differentiation of tumorspheres. Finally, 2 weeks of daily ATRA treatment were sufficient to inhibit gastric tumor progression in vivo, which was associated with a decrease in CD44, ALDH1, Ki67 and PCNA expression in the remaining tumor cells. Administration of ATRA appears to be a potent strategy to efficiently inhibit tumor growth and more importantly to target gastric CSCs in both intestinal and diffuse types of gastric carcinoma.

scholarly journals Targeting Lung Cancer Stem Cells with Antipsychological Drug Thioridazine

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Haiying Yue ◽  
Dongning Huang ◽  
Li Qin ◽  
Zhiyong Zheng ◽  
Li Hua ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Cell Cycle ◽  
Stem Cells ◽  
Cancer Stem Cells ◽  
Cancer Progression ◽  
Cell Cycle Distribution ◽  
Lung Cancer Stem Cells ◽  
Sphere Formation

Lung cancer stem cells are a subpopulation of cells critical for lung cancer progression, metastasis, and drug resistance. Thioridazine, a classical neurological drug, has been reported with anticancer ability. However, whether thioridazine could inhibit lung cancer stem cells has never been studied. In our current work, we used different dosage of thioridazine to test its effect on lung cancer stem cells sphere formation. The response of lung cancer stem cells to chemotherapy drug with thioridazine treatment was measured. The cell cycle distribution of lung cancer stem cells after thioridazine treatment was detected. The in vivo inhibitory effect of thioridazine was also measured. We found that thioridazine could dramatically inhibit sphere formation of lung cancer stem cells. It sensitized the LCSCs to chemotherapeutic drugs 5-FU and cisplatin. Thioridazine altered the cell cycle distribution of LCSCs and decreased the proportion of G0 phase cells in lung cancer stem cells. Thioridazine inhibited lung cancer stem cells initiated tumors growth in vivo. This study showed that thioridazine could inhibit lung cancer stem cells in vitro and in vivo. It provides a potential drug for lung cancer therapy through targeting lung cancer stem cells.

A Novel Camptothecin Derivative 3j Inhibits Nsclc Proliferation Via Induction of Cell Cycle Arrest By Topo I-Mediated DNA Damage

2019 ◽  
Vol 19 (3) ◽  
pp. 365-374 ◽  
Author(s):  
Yang Liu ◽  
Jingyin Zhang ◽  
Shuyun Feng ◽  
Tingli Zhao ◽  
Zhengzheng Li ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Dna Damage ◽  
Cyclin D ◽  
Dna Relaxation ◽  
Cycle Arrest ◽  
H460 Cell ◽  
H1975 Cell

Objective: The aim of this study is to investigate the inhibitory effect of camptothecin derivative 3j on Non-Small Cell Lung Cancer (NSCLCs) cells and the potential anti-tumor mechanisms. Background: Camptothecin compounds are considered as the third largest natural drugs which are widely investigated in the world and they suffered restriction because of serious toxicity, such as hemorrhagic cystitis and bone marrow suppression. Methods: Using cell proliferation assay and S180 tumor mice model, a series of 20(S)-O-substituted benzoyl 7- ethylcamptothecin compounds were screened and evaluated the antitumor activities in vitro and in vivo. Camptothecin derivative 3j was selected for further study using flow cytometry in NSCLCs cells. Cell cycle related protein cyclin A2, CDK2, cyclin D and cyclin E were detected by Western Blot. Then, computer molecular docking was used to confirm the interaction between 3j and Topo I. Also, DNA relaxation assay and alkaline comet assay were used to investigate the mechanism of 3j on DNA damage. Results: Our results demonstrated that camptothecin derivative 3j showed a greater antitumor effect in eleven 20(S)-O-substituted benzoyl 7-ethylcamptothecin compounds in vitro and in vivo. The IC50 of 3j was 1.54± 0.41 µM lower than irinotecan with an IC50 of 13.86±0.80 µM in NCI-H460 cell, which was reduced by 8 fold. In NCI-H1975 cell, the IC50 of 3j was 1.87±0.23 µM lower than irinotecan (IC50±SD, 5.35±0.38 µM), dropped by 1.8 fold. Flow cytometry analysis revealed that 3j induced significant accumulation in a dose-dependent manner. After 24h of 3j (10 µM) treatment, the percentage of NCI-H460 cell in S-phase significantly increased (to 93.54 ± 4.4%) compared with control cells (31.67 ± 3.4%). Similarly, the percentage of NCI-H1975 cell in Sphase significantly increased (to 83.99 ± 2.4%) compared with control cells (34.45 ± 3.9%) after treatment with 10µM of 3j. Moreover, increased levels of cyclin A2, CDK2, and decreased levels of cyclin D, cyclin E further confirmed that cell cycle arrest was induced by 3j. Furthermore, molecular docking studies suggested that 3j interacted with Topo I-DNA and DNA-relaxation assay simultaneously confirmed that 3j suppressed the activity of Topo I. Research on the mechanism showed that 3j exhibited anti-tumour activity via activating the DNA damage response pathway and suppressing the repair pathway in NSCLC cells. Conclusion: Novel camptothecin derivative 3j has been demonstrated as a promising antitumor agent and remains to be assessed in further studies.

scholarly journals STEM-26. SMALL MOLECULE DRUG CBL0137 INHIBITS THE FACT COMPLEX AND SENSITIZES GLIOBLASTOMA CANCER STEM CELLS TO RADIOTHERAPY

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii201-ii202
Author(s):  
Miranda Tallman ◽  
Abigail Zalenski ◽  
Amanda Deighen ◽  
Morgan Schrock ◽  
Sherry Mortach ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cancer Stem Cells ◽  
High Rate ◽  
Orthotopic Model ◽  
In Vivo Models ◽  
Specific Cytotoxicity ◽  
Treatment Paradigm ◽  
Cancer Agent

Abstract Glioblastoma (GBM) is a malignant brain tumor with nearly universal recurrence. GBM cancer stem cells (CSCs), a subpopulation of radio- and chemo-resistant cancer cells capable of self-renewal, contribute to the high rate of recurrence. The anti-cancer agent, CBL0137, inhibits the FACT (facilitates chromatin transcription) complex leading to cancer cell specific cytotoxicity. Here, we show that CBL0137 sensitized GBM CSCs to radiotherapy using both in vitro and in vivo models. Treatment of CBL0137 combined with radiotherapy led to increased DNA damage in GBM patient specimens and failure to resolve the damage led to decreased cell viability. Using clonogenic assays, we confirmed that CBL0137 radiosensitized the CSCs. To validate that combination therapy impacted CSCs, we used an in vivo subcutaneous model and showed a decrease in the frequency of cancer stem cells present in tumors as well as decreased tumor volume. Using an orthotopic model of GBM, we confirmed that treatment with CBL0137 followed by radiotherapy led to significantly increased survival compared to either treatment alone. Radiotherapy remains a critical component of patient care for GBM, even though there exists a resistant subpopulation. Radio-sensitizing agents, including CBL0137, pose an exciting treatment paradigm to increase the efficacy of irradiation, especially by inclusively targeting CSCs.

scholarly journals Targeting specificity of dendritic cells on breast cancer stem cells: in vitro and in vivo evaluations

2015 ◽  
pp. 323 ◽  
Author(s):  
Phuc Pham ◽  
Sinh Nguyen ◽  
Viet Pham ◽  
Ngoc Phan ◽  
Huyen Nguyen ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Stem Cells ◽  
Dendritic Cells ◽  
Cancer Stem Cells ◽  
Breast Cancer Stem Cells

scholarly journals Genome-wide CRISPR-Cas9 screen identified KLF11 as a druggable suppressor for sarcoma cancer stem cells

2021 ◽  
Vol 7 (5) ◽  
pp. eabe3445
Author(s):  
Yicun Wang ◽  
Jinhui Wu ◽  
Hui Chen ◽  
Yang Yang ◽  
Chengwu Xiao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cancer Stem Cells ◽  
Negative Feedback ◽  
Direct Interaction ◽  
Therapy Resistance ◽  
Negative Regulator ◽  
Chemotherapy Response ◽  
Genome Wide

Cancer stem cells (CSCs) are involved in tumorigenesis, recurrence, and therapy resistance. To identify critical regulators of sarcoma CSCs, we performed a reporter-based genome-wide CRISPR-Cas9 screen and uncovered Kruppel-like factor 11 (KLF11) as top candidate. In vitro and in vivo functional annotation defined a negative role of KLF11 in CSCs. Mechanistically, KLF11 and YAP/TEAD bound to adjacent DNA sites along with direct interaction. KLF11 recruited SIN3A/HDAC to suppress the transcriptional output of YAP/TEAD, which, in turn, promoted KLF11 transcription, forming a negative feedback loop. However, in CSCs, this negative feedback was lost because of epigenetic silence of KLF11, causing sustained YAP activation. Low KLF11 was associated with poor prognosis and chemotherapy response in patients with sarcoma. Pharmacological activation of KLF11 by thiazolidinedione effectively restored chemotherapy response. Collectively, our study identifies KLF11 as a negative regulator in sarcoma CSCs and potential therapeutic target.

scholarly journals LncRNA MONC Suppresses the Malignant Phenotype of Endometrial Cancer Stem Cells by Regulating the MiR-636/GLCE Axis

2020 ◽  
Author(s):  
Yibing Li ◽  
Jianing Huo ◽  
Junjian He ◽  
Haining Ma ◽  
Xiaoxin Ma
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Endometrial Cancer ◽  
Cancer Stem Cells ◽  
Suppressive Effect ◽  
Luciferase Assay ◽  
Biological Behavior ◽  
Mesenchymal Transition ◽  
Tumor Suppressive Effect

Abstract Background: Emerging evidence shows that abnormal expression of long non-coding RNA is involved in the occurrence and development of various tumors. LncRNA MONC is abnormally expressed in head and neck squamous cell carcinoma (HNSCC), lung cancer, colorectal cancer, and acute megakaryocytic leukemia, but the biological function and potential regulatory mechanism of MONC in endometrial cancer stem cells (ECSCs) and endometrial cancer cells (ECCs) have not been studied. In this study, we aimed to explore the tumor suppressive effect and mechanism of MONC in regulating ECSCs and ECCs. Methods: The expression of genes was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The expression of proteins was detected by Western blot. The interplay of LncRNA-miRNA-mRNA was verified using the luciferase assay. The growth rate of ECSC spheroids was detected by sphere formation assay. Cell proliferation was detected by CCK-8 assay. The cell invasion was detected by transwell invasion assay. Cell cycle was detected by Cell cycle analysis.Cell apoptosis was detected by the Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) double-staining assay. Animal study was conducted to evaluate the effect of MONC combined with miR-636 on tumor growth in vivo. Results: Low MONC expression in endometrial carcinoma (EC), which directly inhibits the malignant biological behavior of ECSCs and ECCs by directly inhibiting miR-636. Simultaneously, miR-636 may indirectly reduce the expression of MONC. Down-regulation of miR-636 may promote GLCE expression by targeting the 3'-untranslated region (UTR) of the downstream gene GLCE, thereby inhibiting the progression of ECSCs. MONC combined with miR-636 inhibited the Notch signaling pathway and tumor epithelial-to-mesenchymal transition (EMT) process. In addition, we verified the tumor suppressive effect of MONC in nude mice, miR-636 can rescue the tumor suppressive effect of overexpressing MONC, and this effect is more obvious in ECSC. Conclusion: MONC inhibits the malignant phenotypes of ECSCs and ECCs by regulating the miR-636/GLCE axis. The MONC/miR-636/GLCE axis may provide novel treatment avenues for human EC.

scholarly journals The role of ProMyelocytic Leukemia (PML) protein in breast cancer and breast cancer stem cells

2018 ◽  
Author(s):  
Νικολέτα-Νίκη Σαχίνη
Keyword(s):  
Breast Cancer ◽  
Stem Cells ◽  
Cancer Stem Cells ◽  
Promyelocytic Leukemia ◽  
Breast Cancer Stem Cells ◽  
Forkhead Box ◽  
Forkhead Box M1

Ο καρκίνος του μαστού αποτελεί την πιο συχνή μορφή καρκίνου στις γυναίκες. Πρόκειται για μια ετερογενή ασθένεια όσον αφορά το φαινότυπό της και το γενετικό της υπόβαθρο. Καρκινικά χαρακτηριστικά, όπως ο ρυθμός πολλαπλασιασμού, η διεισδυτικότητα, η μεταστατικότητα, η ανθεκτικότητα στα φάρμακα και συγκεκριμένες ογκογόνες μεταλλαγές διαφέρουν μεταξύ περιπτώσεων καρκίνου του μαστού. Τέτοιου είδους ετερογένεια (ενδο-ογκική και δι-ογκική) μεταξύ των όγκων προκύπτει από στοχαστικές γενετικές και επιγενετικές αλλαγές οι οποίες προσδίδουν κληρονομήσιμες φαινοτυπικές και λειτουργικές διαφορές μεταξύ των καρκινικών κυττάρων. Παρ’ όλο που υπάρχουν θεραπευτικά σχήματα που αντιμετωπίζουν επιτυχώς την ασθένεια, η ετερογενής της φύση την καθιστά δύσκολο θεραπευτικό στόχο. Επομένως, η κατανόηση των παθογενετικών και μοριακών μηχανισμών οι οποίοι διέπουν τη νόσο και τους διαφορετικούς υποτύπους της είναι απαραίτητη για την ανακάλυψη νέων φαρμακευτικών στόχων και το σχεδιασμό εξατομικευμένης θεραπείας.Η πρωτεΐνη της προμυελοκυτταρικής λευχαιμίας (PML) περιγράφεται συνήθως ως ογκοκατασταλτικός παράγοντας λόγω των προ-αποπτωτικών, ανασταλτικών του κυτταρικού κύκλου και προγηραντικών ιδιοτήτων της. Συνεπώς, η έκφρασή της απουσιάζει από πρωτοπαθή δείγματα διαφόρων νεοπλασιών, συμπεριλαμβανομένου και του καρκίνου του μαστού. Παρ’ όλα αυτά σε πρόσφατες βιβλιογραφικές αναφορές η PML χαρακτηρίζεται ως ογκογόνος παράγοντας. Συγκεκριμένα, στη χρόνια μυελογενή λευχαιμία, σε γλοιοβλαστώματα και σε ορισμένες περιπτώσεις τριπλά αρνητικού καρκίνου του μαστού η PML εκφράζεται σε υψηλά επίπεδα. Οι προ-ογκογόνες ιδιότητες της PML φαίνεται να σχετίζονται με τη ρύθμιση του κυτταρικού κύκλου των φυσιολογικών και καρκινικών βλαστοκυττάρων αλλά και τη διατήρηση της αυτο-ανανέωσης, μέσω μεταβολικών οδών π.χ. οξείδωση των λιπαρών οξέων. Επομένως, προκύπτει ότι υπό συγκεκριμένες συνθήκες η PML έχει διττό ρόλο στην ογκογένεση εκδηλώνοντας ογκοκατασταλτική ή ογκογόνα δράση.Σκοπός της παρούσας διδακτορικής διατριβής ήταν η αποσαφήνιση των μοριακών και επιγενετικών μηχανισμών με τους οποίους η PML ρυθμίζει τον κυτταρικό πολλαπλασιασμό και τα μονοπάτια αυτό-ανανέωσης στον καρκίνο του μαστού. Τα πειραματικά μας δεδομένα υποστηρίζουν ότι η υπερέκφραση (ΥΕ) της PML αναστέλλει τον πολλαπλασιασμό των κυττάρων και μπλοκάρει τον κυτταρικό κύκλο της τριπλά αρνητικά κυτταρικής σειράς καρκίνου του μαστού, MDA-MB-231. Από την ανάλυση του μεταγραφικού προφίλ αυτών των κυττάρων προκύπτει ότι σημαντικά σηματοδοτικά μονοπάτια και βιολογικές λειτουργίες, όπως και οι ρυθμιστές τους, διαταράσσονται μετά από ΥΕ PML. Ενδιαφέρον παρουσιάζει το γεγονός ότι πολλά από τα γονίδια σχετιζόμενα με τον κυτταρικό πολλαπλασιασμό των οποίων η έκφραση μειώνεται μετά από ΥΕ PML είναι και, θετικά, ρυθμιζόμενοι στόχοι του μεταγραφικού παράγοντα FOXM1 (Forkhead Box M1). Η λειτουργική αλληλεπίδραση της PMLIV με την επικράτεια πρόσδεσης του FOXM1 στο DNA, καθώς και η μείωση του FOXM1 σε επίπεδο mRNA και πρωτεΐνης, υποδεικνύουν ότι η PMLIV είναι ένας καταστολέας του FOXM1. Επιπλέον, δείξαμε ότι η PMLIV YE επηρεάζει το μεταγραφικό πρόγραμμα του μεταγραφικού παράγοντα, FOXO3, o οποίος δρα ανοδικά του FOXM1. Συμπερασματικά, προτείνουμε ότι η PML επηρεάζει την ισορροπία των FOXO3 και FOXM1 μεταγραφικών προγραμμάτων στοχεύοντας διακριτά υποσύνολα γονιδίων. Τα αποτελέσματα μας δείχνουν ότι υψηλά επίπεδα PML μπορούν να επηρεάσουν ταυτόχρονα ποικίλα σηματοδοτικά μονοπάτια στοχεύοντας τον άξονα FOXO3-FOXM1, ο οποίος συνδέει τον κυτταρικό πολλαπλασιασμό (FOXM1) με την απόπτωση, διακοπή του κυτταρικού κύκλου και μηχανισμούς επιβίωσης (FOXO3). Βρήκαμε ακόμη ότι εκτός από τους FOXO3 και FOXM1, η ΥΕ PML στοχεύει κι άλλους καίριους μεταγραφικούς παράγοντες (NFYA,E2F,TBP) με γενικό ρόλο στη μεταγραφή και στον κυτταρικό πολλαπλασιασμό. Συνεπώς, θεωρούμε ότι η PMLIV ΥΕ επιδρά σε σημαντικές βιολογικές διεργασίες μέσω ενός σύνθετου δικτύου μεταγραφικών παραγόντων και επιγενετικών αλλαγών. Αρχικά πειράματα PMLIV YE σε κύτταρα διαφορετικού τύπου καρκίνου του μαστού, τα Τ47D, έδειξαν ότι η PMLIV YE οδηγεί σε αναστολή του κυτταρικού πολλαπλασιασμού, αλλά προκαλεί και διαφορετικές αποκρίσεις σε σχέση με τα MDA-MB-231 κύτταρα.Μελετήσαμε ακόμη το ρόλο της PMLIV σε βλαστικά καρκινικά κύτταρα του μαστού. Παρατηρήσαμε ότι η PMLIV YE μειώνει την ικανότητα σχηματισμού ογκοσφαιρών στην καρκινική σειρά MDA-MB-231, υποδηλώνοντας ότι η PMLIV μειώνει την αυτό-ανανέωση των βλαστικών καρκινικών κυττάρων. Επιπλέον, ερευνήσαμε την επίδραση της PMLIV σε απομονωμένους, με βάση την έκφραση των επιφανειακών δεικτών EpCAM/CD24, υποπληθυσμούς καρκινικών κυττάρων και διαπιστώσαμε ότι η PMLIV YE επηρεάζει με διαφορετικό τρόπο την έκφραση γονιδίων που εμπλέκονται στη διαφοροποίηση του επιθηλίου, στην επιθηλιακή προς μεσεγχυματική μετάβαση, στον πολλαπλασιασμό και στη βλαστικότητα του κάθε υποπληθυσμού, καθώς και την ικανότητα αυτο-ανανέωσης η οποία καθορίστηκε in vitro με τη δοκιμασία σφαιρογένεσης και in vivo με ξενομοσχεύματα. Επομένως, τα αρχικά μας δεδομένα, σε αντίθεση με προηγούμενες αναφορές, υποστηρίζουν την άποψη ότι πολύ ογκογόνοι κυτταρικοί υποπληθυσμοί εμπίπτουν σε περισσότερους από έναν EpCAM/CD24 υπότυπους οι οποίοι επηρεάζονται από την PMLIV με διαφορετικό τρόπο.Συμπερασματικά, η ερευνά μας δίνει πληροφορίες για τη ρύθμιση της ανάπτυξης των όγκων από την PML στον καρκίνο του μαστού. Προσδιορίσαμε τους FOXO3 και FOXM1 σαν αλληλεπιδρώντες παράγοντες της PML, οι οποίοι προσδιορίζουν και το λειτουργικό αποτέλεσμα της PML στο κυτταρικό υπόβαθρο των MDA-MB-231 κυττάρων και αναδείξαμε έναν καινούριο, ρυθμιστικό άξονα στον πολλαπλασιασμό του καρκίνου του μαστού, PML/FOXO3/FOXM1, ο οποίος μπορεί να είναι θεραπευτικός στόχος. Επιπλέον, οι αρχικές παρατηρήσεις στα T47D κύτταρα, ενισχύουν την άποψη περί κυτταρικού υποβάθρου εξαρτώμενης δράσης της PML. Η PMLIV YE επηρέασε πολύ λιγότερο τη δημιουργία σφαιρών, ένδειξη αυτο-ανανέωσης, στα T47D σε σχέση με τα MDA-MB-231. Επιπλέον, συγκρίνοντας το μεταγραφικό προφίλ των δύο κυτταρικών σειρών, οι οποίες αντιπροσωπεύουν διαφορετικούς μοριακούς υποτύπους του καρκίνου του μαστού, μετά από PMLIV YE διαπιστώσαμε ότι η PMLIV επηρεάζει τόσο κοινές αλλά και διακριτές ομάδες γονιδίων. Προτείνουμε ότι η PML ευνοεί τόσο την αναστολή της ανάπτυξης του όγκου όσο και μηχανισμούς επιβίωσης σε διαφορετικό βαθμό, ο οποίος καθορίζεται από το εκάστοτε γενετικό και επιγενετικό υπόβαθρο των κυττάρων.

scholarly journals Transcriptome Profiling of Panc-1 Spheroid Cells with Pancreatic Cancer Stem Cells Properties Cultured by a Novel 3D Semi-Solid System

2018 ◽  
Vol 47 (5) ◽  
pp. 2109-2125 ◽  
Author(s):  
Zhaocong Yang ◽  
Yanfeng Zhang ◽  
Tingting Tang ◽  
Qinhua Zhu ◽  
Wanyue Shi ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Stem Cells ◽  
Cancer Stem Cells ◽  
Culture System ◽  
High Throughput Sequencing ◽  
Semi Solid ◽  
Pancreatic Cancer Stem Cells ◽  
Solid System

Background/Aims: Pancreatic cancer remains one of the deadliest human malignancies, the lethality of which may be attributed to the presence of pancreatic cancer stem cells (PCSCs), a small subpopulation of cells existing within pancreatic tumor with high carcinogenesis. Therefore, it is crucial to establish an efficient enrichment and culture system of PCSCs and identify the key genes involved in the regulation of PCSCs. The three-dimensional (3D) liquid suspension mammosphere culture system has been established for enrichment and culture of PCSCs in vitro as the cell spheres are likely to originate from individual cell clone, but it has been challenged because the cell spheroids could be a result of cell aggregation. Methods: We optimized the existing culture system by adding methylcellulose to create a 3D semi-solid system which prevented the non-specific aggregation. Then we identified the CSC properties of Panc-1 spheroid cells cultured by this system by detecting the genes associated with stemness and by evaluation of the tumorigenicity in vitro and in vivo through invasion, migration and xenograft experiments methods. Subsequently, we performed high-throughput sequencing (HTS) of the Panc-1 spheroid cells. Results: We confirmed the PCSCs properties and high malignancy of the Panc-1 spheroid cells enriched by our novel 3D semi-solid system both in vitro and in vivo. Hundreds of mRNA, microRNA (miRNA) and dozens of long non-coding RNA (LncRNA) were identified to be differentially regulated in PCSCs-like Panc-1 spheroid cells compared with their parental cells by HTS. Conclusions: Our results demonstrate an efficient enrichment and culture system for Panc-1 spheroid cells with the PCSCs properties. The differentially expressed genes and their targets identified by the HTS of the Panc-1 spheroid cells can serve as new potential biomarkers for pancreatic cancer diagnosis and targeted therapy.

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