scholarly journals Exosomal miR-331 from Chemoresistant Osteosarcoma Cells Induced Chemoresistance Through Autophagy

2021 ◽  
Author(s):  
Aiqing Zhao ◽  
Yan bin Zhao ◽  
Wanlin Liu ◽  
Wei Feng ◽  
Wenhua Xing ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Resistant Cell ◽  
Drug Resistant ◽  
Rt Pcr ◽  
Chemotherapeutic Resistance ◽  
Acquire Drug Resistance ◽  
And Migration ◽  
Drug Resistant Cell ◽  
Resistant Cells

Abstract Background Osteosarcoma (OS) is a highly malignant tumor. Improving chemotherapeutic resistance is very important to improve the survival rate of OS. Exosomes and microRNAs (MiRNA) play important roles in the mechanism of chemotherapeutic resistance transmission. More and more researches focus the mechanism of miRNAs carried by exosomes in the transmission of chemotherapeutic resistance of OS. This study focused on exploring the mechanism of exosomal miR-331 in the transmission of chemoresistance in OS. Methods We cultured OS drug-resistant cells and extracted exosomes of these cells. The secretion and uptake of exosomes in OS drug-resistant cells and OS cells (OSCs) were confirmed by fluorescence tracking assay and transwell experiments. The differential expression of microRNA-331 (miR-331) in exosomes of OS resistant and OS cells was investigated by RT-PCR. The effects of drug-resistant exosomes on proliferation and migration of OS cells were determined by MTT assay and scratches assay. MDC staining, RT-PCR, and Western blot were used to detect the role of autophagy which regulated by drug-resistant cell-derived exosom-miR-331. Results We found that the expression difference of miR-331 between drug-resistant cells of MG63 and HOS cell lines and tumor cells was the most significant. Drug resistant OSCs secreted exosomes and were ingested by OSCs, which then promoted OSCs to acquire drug resistance. In addition, exosomes secreted by drug-resistant OSCs promote drug resistance by carrying miRNAs. Interestingly, inhibition of miRNA resulted in reduced drug resistance transmission of exosomes. Finally, we found that the exosomes secreted by drug-resistant OSCs could induce autophagy of OSCs by carrying miR-331, thus making OSCs acquire drug resistance. Inhibition of miR-331 can effectively improve drug resistance of OSCs. Conclusions Chemoresistant OSCs-derived exosomes promote the transmission of drug resistance by carrying miR-331 and inducing autophagy. Inhibition of miR-331 could effectively alleviate drug resistance of OSCs.

scholarly journals Exosomal miR-331 from Chemoresistant Osteosarcoma Cells induced Chemoresistance through Autophagy

2021 ◽  
Author(s):  
Aiqing Zhao ◽  
Yanbin Zhao ◽  
Wanlin Liu ◽  
Wei Feng ◽  
Wenhua Xing ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Resistant Cell ◽  
Drug Resistant ◽  
Rt Pcr ◽  
Chemotherapeutic Resistance ◽  
Acquire Drug Resistance ◽  
And Migration ◽  
Drug Resistant Cell ◽  
Resistant Cells

Abstract BackgroundOsteosarcoma (OS) is a highly malignant tumor. Improving chemotherapeutic resistance is very important to improve the survival rate of OS. Exosomes and microRNAs (MiRNA) play important roles in the mechanism of chemotherapeutic resistance transmission. More and more researches focus the mechanism of miRNAs carried by exosomes in the transmission of chemotherapeutic resistance of OS. This study focused on exploring the mechanism of exosomal miR-331 in the transmission of chemoresistance in OS.MethodsWe cultured OS drug-resistant cells and extracted exosomes of these cells. The secretion and uptake of exosomes in OS drug-resistant cells and OS cells (OSCs) were confirmed by fluorescence tracking assay and transwell experiments. The differential expression of microRNA-331 (miR-331) in exosomes of OS resistant and OS cells was investigated by RT-PCR. The effects of drug-resistant exosomes on proliferation and migration of OS cells were determined by MTT assay and scratches assay. MDC staining, RT-PCR, and Western blot were used to detect the role of autophagy which regulated by drug-resistant cell-derived exosom-miR-331.ResultsWe found that the expression difference of miR-331 between MG63/CDDP and MG63 was the most significant. Drug resistant OSCs secreted exosomes and were ingested by OSCs, which then promoted OSCs to acquire drug resistance. In addition, exosomes secreted by drug-resistant OSCs promote drug resistance by carrying miRNAs. Interestingly, inhibition of miRNA resulted in reduced drug resistance transmission of exosomes. Finally, we found that the exosomes secreted by drug-resistant OSCs could induce autophagy of OSCs by carrying miR-331, thus making OSCs acquire drug resistance. Inhibition of miR-331 can effectively improve drug resistance of OSCs.ConclusionsChemoresistant OSCs-derived exosomes promote the transmission of drug resistance by carrying miR-331 and inducing autophagy. Inhibition of miR-331 could effectively alleviate drug resistance of OSCs.

scholarly journals Drug-resistant cancer cell-derived exosomal EphA2 promotes breast cancer metastasis via the EphA2-Ephrin A1 reverse signaling

2021 ◽  
Vol 12 (5) ◽  
Author(s):  
Zicong Gao ◽  
Xingxing Han ◽  
Yuying Zhu ◽  
He Zhang ◽  
Ran Tian ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Drug Resistance ◽  
Cancer Cells ◽  
Cancer Progression ◽  
Cancer Metastasis ◽  
Resistant Cell ◽  
Breast Cancer Progression ◽  
Drug Resistant ◽  
Drug Resistant Cell ◽  
Resistant Cells

AbstractTumor metastasis induced by drug resistance is a major challenge in successful cancer treatment. Nevertheless, the mechanisms underlying the pro-invasive and metastatic ability of drug resistance remain elusive. Exosome-mediated intercellular communications between cancer cells and stromal cells in tumor microenvironment are required for cancer initiation and progression. Recent reports have shown that communications between cancer cells also promote tumor aggression. However, little attention has been regarded on this aspect. Herein, we demonstrated that drug-resistant cell-derived exosomes promoted the invasion of sensitive breast cancer cells. Quantitative proteomic analysis showed that EphA2 was rich in exosomes from drug-resistant cells. Exosomal EphA2 conferred the invasive/metastatic phenotype transfer from drug-resistant cells to sensitive cells. Moreover, exosomal EphA2 activated ERK1/2 signaling through the ligand Ephrin A1-dependent reverse pathway rather than the forward pathway, thereby promoting breast cancer progression. Our findings indicate the key functional role of exosomal EphA2 in the transmission of aggressive phenotype between cancer cells that do not rely on direct cell–cell contact. Our study also suggests that the increase of EphA2 in drug-resistant cell-derived exosomes may be an important mechanism of chemotherapy/drug resistance-induced breast cancer progression.

Identification of Chemotherapy-Resistant Mechanisms in Childhood T-ALL Cell Lines: Correlation with Genetic Profiles in Patients Having Early Treatment Failure.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4376-4376
Author(s):  
David A. Estes ◽  
Debbie M. Lovato ◽  
Hadya M. Khawaja ◽  
Stuart S. Winter ◽  
Richard S. Larson
Keyword(s):  
Drug Resistance ◽  
T Cell ◽  
Treatment Failure ◽  
Cell Lines ◽  
Drug Sensitivity ◽  
Fold Increase ◽  
Resistant Cell ◽  
Drug Resistant ◽  
Drug Resistant Cell ◽  
Resistant Cells

Abstract Acquisition of drug resistance in childhood T-cell acute lymphoblastic leukemia (T-ALL) is a major cause of treatment failure. Drug resistant cell lines are effective tools for elucidating mechanisms of acquired drug resistance. In this study, we developed novel drug resistant cell lines that could be employed to identify mechanisms of drug resistance. We established three new drug resistant T-ALL cell lines: 2 resistant to L-asparaginase (L-asp), and one resistant to daunorubicin (DNR). The DNR-resistant line (Jurkat) acquired a 117-fold increase in EC50 to DNR, from 20.6 nM to 2.4 μM, while the L-asp resistant cells (Jurkat and Sup T1) showed respective increases in resistance of 320-fold (0.003 IU/mL to 0.962 IU/mL) and 29-fold (0.042 IU/mL to 1.22 IU/mL). The DNR resistant cell line acquired a multidrug resistant phenotype, showing 310 and 120-fold increase in resistance to vincristine and prednisolone, respectively. Resistance to L-asp was unchanged. Microarray analysis showed that ABCB1 (MDR1, P-glycoprotein) was significantly upregulated (567-fold) in DNR resistant cells. siRNA experiments that reduced ABCB1 mRNA levels by 74% restored DNR sensitivity. In L-asp resistant T-ALL cells (Jurkat and Sup T1), two notable genes were upregulated, asparagine synthetase (ASNS) which catalyzes synthesis of asparagine (41-fold and 1.5-fold) and argininosuccinate synthase (ASS)(32-fold and 6.5-fold), respectively. Reduction of ASNS with siRNA restored drug sensitivity in both cell lines. Interestingly, siRNA suppression of ASS in conjunction with ASNS achieved an exaggerated restoration of drug sensitivity compared to ASNS alone. We next examined the microarray profiles of drug resistant cells with those of 86 T-ALL patients; of which 8 failed induction (IF). Interestingly, these 3 key genes are upregulated in 25–62% of IF cases. Although ABCB1 overexpression has been shown to be a mechanism of DNR resistance in many cancers, there is a paucity of resistant T-ALL cell lines to adequately model the effect of stage of differentiation and genetic heterogeneity underlying drug resistance in T-ALL. With the establishment of 3 new T-ALL cell lines in this report, there exist 5 T-ALL drug resistant cell lines, representing the spectrum of T-cell differentiation (pre-T, cortical T, and mature T cells). Finally, this is the first report of the potential contribution of ASS in addition to ASNS to L-asp resistance in leukemia cells.

scholarly journals Drug-resistant cancer cell-derived exosomal EphA2 promotes breast cancer metastasis via the EphA2-Ephrin A1 reverse signaling

2020 ◽  
Author(s):  
Zicong Gao ◽  
Xingxing Han ◽  
Yuying Zhu ◽  
He Zhang ◽  
Ran Tian ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Drug Resistance ◽  
Cancer Cells ◽  
Cancer Progression ◽  
Breast Cancer Cells ◽  
Resistant Cell ◽  
Migration And Invasion ◽  
Drug Resistant ◽  
Resistant Cells

Abstract Background: The failure of chemotherapy is accompanied by the emergence of drug resistance and tumor relapse. Tumor metastasis induced by drug resistance is a major challenge in successful cancer treatment. Nevertheless, the mechanisms underlying the pro-invasive and metastatic ability of drug resistance remain elusive. Exosome-mediated intercellular communications between cancer cells and stromal cells in tumor microenvironment are required for cancer initiation and progression. Recent reports have shown that communications between cancer cells also promote tumor aggression. However, little attention has been regarded on this aspect. In this study, we aimed to investigate the mechanisms of exosomes derived from drug-resistant cells in regulating the invasion and metastasis of sensitive breast cancer cells.Methods: Exosomes isolated from drug-resistant breast cancer cells and their parental cells were used to treat breast cancer cells, and then the migration and invasion abilities were examined. The tandem mass tag (TMT)-based quantitative proteomic method was carried out to identify key molecules that regulate cancer aggressiveness. Lentivirus-mediated shRNAs, overexpression, point mutation, truncation mutation, Western blotting, tumor xenograft mice models, and in vivo breast cancer metastatic models were used to investigate the functional role of EphA2 on the invasion and metastatic potential of breast cancer cells.Results: We demonstrated that drug-resistant cell-derived exosomes promoted the migration and invasion of sensitive breast cancer cells. Quantitative proteomic analysis showed that EphA2 was rich in exosomes from drug-resistant cells. Exosomal EphA2 conferred the invasive/metastatic phenotype transfer from drug-resistant cells to sensitive cells. In addition, we provided considerable evidence that exosomal EphA2 activated ERK1/2 signaling through the ligand Ephrin A1-dependent reverse pathway rather than the forward pathway, thereby promoting breast cancer progression. Conclusions: Our findings indicate the key functional role of exosomal EphA2 in the transmission of aggressive phenotype between cancer cells that do not rely on direct cell–cell contact. Our study also suggests that the increase of EphA2 in drug-resistant cell-derived exosomes may be an important mechanism of chemotherapy/drug resistance-induced breast cancer progression.

scholarly journals The Ribosomal Protein L28 Gene Is Involved in Sorafenib Resistance in Hepatocellular Carcinoma

2021 ◽  
Author(s):  
Yi Shi ◽  
Xiaojiang Wang ◽  
Qiong Zhu ◽  
Gang Chen
Keyword(s):  
Hepatocellular Carcinoma ◽  
Drug Resistance ◽  
Hepg2 Cells ◽  
Cell Model ◽  
Resistant Cell ◽  
Drug Resistant ◽  
Resistant Gene ◽  
Key Genes ◽  
Resistant Cells

Abstract Background: Sorafenib is the first molecular-targeted drug for the treatment of advanced hepatocellular carcinoma (HCC). However, its treatment efficiency decreases after a short period of time because of the development of drug resistance. This study investigates the role of key genes in regulating sorafenib-resistance in hepatocellular carcinoma and elucidates the mechanism of drug resistance. Methods: The HCC HepG2 cells were used to generate a sorafenib-resistant cell model by culturing the cells in gradually increasing concentration of sorafenib. RNA microarray was applied to profile gene expression and screen key genes associated with sorafenib resistance. Specific targets were knockdown in sorafenib-resistant HepG2 cells for functional studies. The HCC model was established in ACI rats using Morris hepatoma3924A cells to validate selected genes associated with sorafenib resistance in vivo. Results: The HepG2 sorafenib-resistant cell model was successfully established. The IC50 of sorafenib was 9.988mM in HepG2 sorafenib-resistant cells. A total of 35 up-regulated genes were detected by expression profile chip. High-content screening technology was used and a potential drug-resistant gene RPL28 was filtered out. After knocking down of RPL28 in HepG2 sorafenib-resistant cells, the results of cell proliferation and apoptosis illustrated that RPL28 is the key drug-resistant gene in the cells. Furthermore, it was found that both RNA and protein expression of RPL28 increased in HepG2 sorafenib-resistant specimens of Morris Hepatoma rats. In addition, the expression of functional proteins Ki-67 increased in sorafenib-resistant cells. Conclusion: Our study suggested that RPL28 was a key gene for sorafenib resistance in HCC both in vitro and in vivo.

scholarly journals Long Non-coding RNA MAFG-AS1 Promotes Cell Proliferation, Migration, and EMT by miR-3196/STRN4 in Drug-Resistant Cells of Liver Cancer

2021 ◽  
Vol 9 ◽  
Author(s):  
Tianming Chen ◽  
Bin Huang ◽  
Yaozhen Pan
Keyword(s):  
Colorectal Cancer ◽  
Drug Resistant ◽  
Non Coding Rna ◽  
Different Types ◽  
Non Coding Rnas ◽  
Novel Target ◽  
And Migration ◽  
Long Non Coding Rna ◽  
Resistant Cells

Long non-coding RNAs (lncRNAs) have been shown to participate in the development and progression of several different types of cancer. Past studies indicated that lncRNA MAFG-antisense 1 (AS1) promotes colorectal cancer. However, the role of MAFG-AS1 in hepatocellular carcinoma (HCC) remains unclear. The aim of the present study is to examine the effect of lncRNA MAFG-AS1 on drug resistance HCC. The results indicated that MAFG-AS1 is upregulated in drug-resistant cells. Further, MAFG-AS1 promotes growth and migration of HCC by upregulating STRN4 through absorbing miR-3196. Thus, LncRNA MAFA-AS1 may become a novel target to treat HCC patients.

scholarly journals Understanding the Role of Extracellular Vesicles in Lenalidomide-Resistance Multiple Myeloma

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 1887-1887
Author(s):  
Tomofumi Yamamoto ◽  
Nobuyoshi Kosaka ◽  
Ochiya Takahiro ◽  
Yutaka Hattori
Keyword(s):  
Bone Marrow ◽  
Drug Resistance ◽  
Cell Line ◽  
Cell Lines ◽  
Parental Cell ◽  
Resistant Cell ◽  
Bone Marrow Microenvironment ◽  
Drug Resistant ◽  
Apoptosis Assay ◽  
Drug Resistant Cell

Abstract Multiple myeloma (MM) is a plasma cell malignancy that develops by the accumulation of various genetic abnormalities. In recent years, the prognosis of MM has improved by the development of novel drugs including immunomodulatory drugs (IMiDs), proteasome inhibitors, and antibody drugs. However, MM cells acquired drug resistance by long-term exposure to these therapeutic drugs. MM is a multiclonal disease, and various clone subtypes develop within the bone marrow microenvironment. It has been suggested that drug resistant phenotype could transmit from resistant clones to sensitive ones, and also to immune cells, or mesenchymal stem cells, resulting in the change of bone marrow microenvironment suitable for MM cell survival. It has been shown that extracellular vesicles (EVs) are one of the means of information transmission. EVs are secreted from almost all cells, and the amount of EV secretion is particular high from cancer cells. It was already known that cancer-derived EVs transmitted information associated with cancer progressions such as angiogenesis, metastasis, and drug resistance to the surrounding cells. Thus, EVs were proposed to play an important role in acquisition of drug resistance even though the mechanisms have not been fully understood in MM. In order to understand the mechanism of drug resistance in MM mediated by EVs, lenalidomide resistant cell lines were established by long-term exposure of lenalidomide. The amount of EVs was measured by ExoScreen, which is ultra-sensitive detection method of EVs by measuring surface protein of EVs, such as, CD9 and CD63, and by the nanoparticle tracking analysis. We found that lenalidomide resistant cell lines in KMS21R, KMS27R and KMS34R cell lines secreted about twice more EVs than their parental cell lines, and the amount of EV secretion was correlated with the drug sensitivity of lenalidomide. Suppression of EV secretion by knockdown of TSG101, which is known for EV secretion-associated protein, did not affect lenalidomide resistance. We could suppress the EV secretion to two-thirds, however cell proliferation and caspase activity were not change. From these results, we postulated the two possibilities; 1) EV secretion pathway other than TSG101 is associated with drug resistance via EVs; 2) EV derived from lenalidomide resistant cells can affect the cells exist in bone marrow microenvironment. From these hypotheses, we have done the following experiments. Firstly, to identify the genes which involved in EV secretion pathway associated with drug resistance, RNA sequence among the drug-resistant cell lines and their parental cell lines was performed. Drug resistant cell lines had some genetic abnormality, for instance immune system or angiogenesis. Now, we are detecting the EV secretion associated genes in drug resistant cell lines. Secondly, EV derived from the drug-resistant cell lines and EV derived from the parent cell lines were added to drug sensitive MM cell lines, then lenalidomide is added after 24hr. The cell proliferation and apoptosis assay were evaluated after 48hr. EV derived from the drug-resistant cell lines in KMS34R cell line significantly inhibited cell death measured by MTS assay and apoptosis assay compared with those from the drug sensitive KMS21 and KMS34 cell lines. EVs from KMS34R cell line, which is the most progressed cell line we established, could more transmit drug resistance than those from other cell lines. These results suggested that drug resistance was transmitted from drug-resistant cell lines to non-resistant cell lines via EVs. Now, we are analyzing the component of EV from drug-resistant MM cells by proteome analysis to identify the molecules associated with lenalidomide resistance in MM. In addition, we are investigating the molecules which associated with the secretion of EVs from drug-resistance MM. These results prompted us to hypothesize that attenuating the function of a molecule responsible for EV secretion could lead to the inhibition of cancer development such as drug resistance. It is expected that EVs will be novel therapeutic targets in refractory or relapsed MM. Disclosures Hattori: IDAC inc.: Research Funding; Takeda: Research Funding.

scholarly journals Restoration of Mir-138 Expression By Epigenetic Modulation Overcomes Drug Resistance in Multiple Myeloma Cells

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 2060-2060
Author(s):  
Nasrin Rastgoo ◽  
Jahangir Abdi ◽  
Lun Zhang ◽  
Minjing Wang ◽  
Hong Chang
Keyword(s):  
Drug Resistance ◽  
Chromatin Remodeling ◽  
Promoter Region ◽  
Feedback Loop ◽  
Expression Level ◽  
Drug Resistant ◽  
H3k27me3 Level ◽  
Ezh2 Expression ◽  
Resistant Cells

Abstract Multiple myeloma (MM) is a plasma cell malignancy characterized by highly heterogeneous genomic alteration including frequent changes in chromatin remodeling enzymes. The recent studies showed that deregulation of epigenetics mechanisms play an essential role in MM development. In particular, overexpression of enhancer of zeste homologue 2 (EZH2), the catalytic subunit of Polycomb Complex 2 (PRC2), has been shown in several cancers including MM. However, the detailed function of EZH2 histone methyltransferase in drug resistance of MM has not yet been elucidated. This study sought to investigate the role of histone modification changes in drug resistance of MM. First, we checked if the expression level of the epigenetic modulating enzyme, EZH2, changed in drug resistant cells (8226-R5 and MM1R) in comparison to parental sensitive cells (8226 and MM1S). We found that EZH2 expression was upregulated in 8226-R5 and MM1R cells. In addition, the global H3K27me3 level, a silencing histone modification mark induced by EZH2, was dramatically increased in the resistant MM cells. To support our finding, we analyzed some publically available gene expression datasets (GSE 6477 and GSE26760) and found that the EZH2 expression level was significantly upregulated in relapsed patients in comparison to newly diagnosed MM patients. It also revealed that overexpression of EZH2 correlated with the progression of the disease and poor survival of the MM patients. We next examined the functional effect of EZH2 inhibition on resistant cells using a novel selective EZH2 inhibitor (EPZ-6438) which turned out to significantly inhibit proliferation of MM cells by MTT assay. Furthermore, combination of EPZ-6438 with bortezomib revealed synergistic cytotoxicity effects on 8226-R5 and MM1R cells. In parallel, global H3K27me3 level in MM cells was reduced by EPZ-6438 further confirming its specific functionality. To explore whether miRNAs are associated with EZH2 overexpression in drug resistance, we used several target scan algorithms and identified mir-138, a tumor suppressor miRNA, to potentially target EZH2. We found that mir-138 expression level was downregulated in 8226-R5 and MM1R relative to 8226 and MM1S, respectively. To investigate if the reduction is due to overexpression of EZH2, we performed H3k27me3 chip on the promoter region of mir-138 gene in MM cell lysates. The chip qPCR showed that occupancy of H3K27me3 in the promoter region of mir-138 was increased in 8226-R5 and MM1R cells compared to 8226 and MM1s cells leading to repression of mir-138 gene transcription in the resistant cells.As a result, we concluded that reduction of mir-138 expression level in the resistant MM cells is due to chromatin remodeling changes in its promoter region and identified mir-138 gene as a target of EZH2. To further demonstrate the role of EZH2 in mir-138 expression, 8226-R5 and MM1R were treated with EZH2 inhibitor and mir-138 expression level was compared to untreated cells. We found a significant upregulation of mir-138 expression in EPZ-6438 treated cells compared with untreated controls. The chip qPCR results also confirmed that the inhibition of EZH2 decreased H3K27me3 occupancy on the mir-138 promoter region which indicated that epigenetic silencing is mechanism underlying miR-138 suppression in drug resistant cells. Furthermore, we overexpressed mir-138 in 8226-R5 and MM1R cells and treated them with bortezomib to assess cell viability using MTT assay. Overexpression of mir-138 sensitized 8226-R5 and MM1R cells to bortezomib. We also observed that overexpression of mir-138 reduced EZH2 expression level. These results suggest a negative feedback loop between miR-138 and EZH2 in our model system. Altogether, these findings indicate that downregulation of miR-138, likely due to EZH2 overexpression, contributes to drug resistance of MM and the restoration of mir-138 expression by epigenetics modulation re-sensitizes drug resistant MM cells to anti myeloma drugs. We report, for the first time, that miR-138 is involved in a negative feedback loop with EZH2 in MM cells and provide a framework for targeting EZH2 or restoration of mir-138 as novel strategies for the treatment of MM. Disclosures No relevant conflicts of interest to declare.

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