scholarly journals MicroRNA-139-5p Suppresses Cell Malignant Behaviors in Breast Cancer through Targeting MEX3A

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Jian Chu ◽  
Tangya Li ◽  
Lei Li ◽  
Huiwen Fan
Keyword(s):  
Breast Cancer ◽  
Cell Growth ◽  
Western Blotting ◽  
Underlying Mechanism ◽  
Luciferase Reporter ◽  
Anticancer Effect ◽  
Promoting Effect ◽  
Qrt Pcr ◽  
Apoptosis Level

Objective. The study was designed to evaluate the underlying mechanism of microRNA-139-5p in breast cancer (BC). Methods. Expression statuses of microRNA-139-5p and MEX3A were measured by qRT-PCR and western blotting. The anticancer effect of microRNA-139-5p in vitro was tested by a set of assays. Interaction between microRNA-139-5p and MEX3A was validated by dual-luciferase detection. Results. MicroRNA-139-5p expression in BC cells was obviously low, while MEX3A was significantly overexpressed. MicroRNA-139-5p restrained proliferative, invasive, and migratory abilities of BC cells and increased apoptosis level of BC cells, while MEX3A exerted a promoting effect on BC cell growth. Dual-luciferase reporter detection confirmed that microRNA-139-5p bound to MEX3A 3 ′ -UTR. Conclusions. MicroRNA-139-5p inhibited the development of BC by targeting MEX3A. MicroRNA-139-5p/MEX3A may be a target for BC therapy.

scholarly journals Knockdown of lncRNA HOTTIP Inhibits Retinoblastoma Progression by Modulating the miR-101-3p/STC1 Axis

2021 ◽  
Vol 20 ◽  
pp. 153303382199783
Author(s):  
XiangWen Yuan ◽  
Zhaoyan Sun ◽  
Congxian Cui
Keyword(s):  
Cell Proliferation ◽  
Western Blotting ◽  
Underlying Mechanism ◽  
Luciferase Reporter ◽  
Luciferase Reporter Gene ◽  
Qrt Pcr ◽  
Gene Assay ◽  
Proliferation And Apoptosis ◽  
Y79 Cells

Objective: Retinoblastoma (RB) is a frequent eye cancer in children. Long non-coding RNA (LncRNA) HOXA transcript at the distal tip (HOTTIP) is aberrantly expressed in cancer tissues. This study explores the underlying mechanism of lncRNA HOTTIP in RB. Methods: HOTTIP expression in normal retinal cells and RB cell lines was detected using qRT-PCR. The proliferation of RB cells was measured using CCK-8 and EdU assays, and apoptosis was detected using flow cytometry and Western blotting after the transfection of si-HOTTIP into Y79 cells and pc-HOTTIP into HXO-RB-44 cells. The target relationships between HOTTIP and miR-101-3p, and miR-101-3p and STC1 were predicted by bioinformatics website and verified using dual-luciferase reporter gene assay. The binding of HOTTIP and miR-101-3p was verified using RNA pull-down assay. STC1 mRNA and protein in RB cells were measured using qRT-PCR and Western blotting. Moreover, si-HOTTIP and in-miR-101-3p/in-NC, and si-HOTTIP and pc-STC1/pcDNA were co-transfected into Y79 cells respectively to evaluate cell proliferation and apoptosis. Xenograft study was conducted, and Ki67-positive expression was detected using immunohistochemical staining. Results: HOTTIP expression was promoted in RB tissues and cells. Downregulation of HOTTIP inhibited proliferation and promoted apoptosis of Y79 cells, while upregulation of HOTTIP promoted proliferation and inhibited apoptosis of HXO-RB-44 cells. There were target relationships between HOTTIP and miR-101-3p, and miR-101-3p and STC1. Inhibition of miR-101-3p or overexpression of STC1 reversed the effect of si-HOTTIP on the proliferation and apoptosis of RB cells. Xenograft study showed that knockdown of HOTTIP suppressed the growth of RB in vitro. Conclusion: It could be concluded that HOTTIP sponged miR-101-3p to upregulate STC1 expression, thereby promoting RB cell proliferation and inhibiting apoptosis.

scholarly journals Long Non-coding RNA TMEM220-AS1 Suppressed Hepatocellular Carcinoma by Regulating the miR-484/MAGI1 Axis as a Competing Endogenous RNA

2021 ◽  
Vol 9 ◽  
Author(s):  
Cong Cao ◽  
Jun Li ◽  
Guangzhi Li ◽  
Gaoyu Hu ◽  
Zhihua Deng ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Western Blotting ◽  
Pdz Domain ◽  
Cell Counting ◽  
Luciferase Reporter ◽  
Qrt Pcr ◽  
Low Levels ◽  
Hcc Cells

Long non-coding RNAs (lncRNAs) have a considerable regulatory influence on multiple biological processes. Nevertheless, the role of TMEM220-AS1 in hepatocellular carcinoma (HCC) remains unclear. We used The Cancer Genome Atlas (TCGA) database to analyze the differentially expressed lncRNAs. qRT-PCR was used to verify the results for a large population. The in vitro effects of TMEM220-AS1 on HCC cells were determined using Cell Counting Kit-8 (CCK-8), 5-ethynyl-2’-deoxyuridine (EdU), flow cytometry, and Transwell assays in HCC cells. We used qRT-PCR and western blotting to identify the epithelial-mesenchymal transition (EMT). Moreover, we performed bioinformatics analysis, western blotting, dual luciferase reporter gene assay, RNA pull-down, and RNA binding protein immunoprecipitation (RIP) to investigate the underlying molecular mechanisms of TMEM220-AS1 function. Finally, the function of TMEM220-AS1 was verified in vivo. The results showed that TMEM220-AS1 was expressed at considerably low levels in HCC. It was demonstrated that malignant phenotypes and EMT of HCC cells were promoted by the knock down of TMEM220-AS1 both in vivo and in vitro. TMEM220-AS1, which was detected primarily in the cytoplasm, functioned as an miRNA sponge to bind miR-484 and promote the level of membrane-associated guanylate kinase, WW, and PDZ domain containing 1 (MAGI1), thereby curbing the malignant phenotypes of HCC cells. In conclusion, low levels of TMEM220-AS1 promote proliferation and metastasis through the miR-484/MAGI1 axis in HCC.

scholarly journals LncRNA EGFR-AS1 Regulates Breast Cancer Cells Function and Sensitivity to Docetaxel Via the miR-149-5p/ELP5 Axis

2021 ◽  
Author(s):  
Shiping Li ◽  
Xiaoyi Mi ◽  
Mingfang Sun ◽  
Jie Zhang ◽  
Miaomiao Hao ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Western Blotting ◽  
Breast Cancer Cells ◽  
Breast Cancer Tissue ◽  
Luciferase Reporter ◽  
Cancer Tissue ◽  
Cancer Tissues

Abstract Background: Recently, an increasing number of studies have focused on investigating long non-coding RNAs (lncRNAs) and their role in regulating the progression of various cancer types. However, the biological effects and underlying mechanisms of EGFR-AS1, a typical lncRNA, remain largely unclear in breast cancer.Methods: Differential expression of EGFR-AS1 in breast cancer tissue was analyzed using an integrative database and verified in breast cancer tissue samples and cells via real-time PCR analysis and western blotting analysis. The tumor promoter role of EGFR-AS1 in breast cancer cells was determined through MTT, EDU analysis, colony formation and transwell assays,and the effect of EGFR-AS1 on docetaxel drug sensitivity was examined. We then performed bioinformatic analysis and the dual-luciferase reporter assay to identify the binding sites of EGFR-AS1/miR-149-5p and miR-149-5p/ELP5. Results from western blotting and biological function studies provided insights into whether the EGFR-AS1/miR-149-5p/ELP5 axis regulates breast cancer development in vitro and in vivo. Results: EGFR-AS1 is upregulated in breast cancer tissues and cells and promotes the progression of breast cancer cells both in vitro and in vivo. Moreover, miR-149-5p is downregulated in breast cancer tissues and cell lines. Mechanistically, EGFR-AS1 regulates ELP5 levels by sponging miR-149-5p, thereby affecting cell progression and promoting epithelial-to-mesenchymal transition. Hence, the EGFR-AS1/miR-149-5p/ELP5 axis is involved in breast cancer proliferation, migration, invasion, and resistance to the chemotherapeutic drug, docetaxel, in breast cancer cells. Conclusions: EGFR-AS1 sponges miR-149-5p to affect the expression level of ELP5 ultimately acting as a new tumor promotor in breast cancer. This study provides novel insights into diagnostic and docetaxel-related chemotherapy targets for breast cancer.

scholarly journals MSC-AS1 knockdown inhibits cell growth and temozolomide resistance by regulating miR-373-3p/CPEB4 axis in glioma through PI3K/Akt pathway

2020 ◽  
Author(s):  
Chong Li ◽  
Shiyu Feng ◽  
Ling Chen
Keyword(s):  
Cell Growth ◽  
Glioma Cells ◽  
Cell Counting ◽  
Luciferase Reporter ◽  
Akt Pathway ◽  
Promoting Effect ◽  
Element Binding Protein ◽  
Glioma Growth

Abstract Long non-coding RNAs (lncRNAs) have been widely reported to regulate the development and chemoresistance of a variety of tumors. Temozolomide (TMZ) is a first-line chemotherapy for treatment of glioma. However, the effect and the regulatory mechanism of lncRNA MSC-AS1 (MSC-AS1) in TMZ-resistant glioma remain unrevealed. Levels of MSC-AS1, microRNA-373-3p (miR-373-3p), and cytoplasmic polyadenylation element binding protein 4 (CPEB4) were determined by quantitative real-time polymerase chain reaction (qRT-PCR). All protein expression was detected by western blot. Cell viability and the half maximal inhibitory concentration (IC50) value of TMZ was assessed by cell counting kit-8 (CCK-8) assay. Cell cloning ability and apoptosis were examined by colony formation and flow cytometry assays, respectively. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were performed to verify the correlation between miR-373-3p and MSC-AS1 or CPEB4. The xenograft models were established to determine the effect of MSC-AS1 in vivo. MSC-AS1 was up-regulated in TMZ-resistant glioma tissues and cells, and glioma patients with high MSC-AS1 expression tend to have lower overall survival rate. MSC-AS1 suppression reduced the IC50 value of TMZ and proliferation, promoted apoptosis and TMZ sensitivity, and affected PI3K/Akt pathway in TMZ-resistant glioma cells. MSC-AS1 acted as miR-373-3p sponge, and miR-373-3p directly targeted CPEB4. Silencing miR-373-3p reversed the promoting effect of MSC-AS1 or CPEB4 knockdown on TMZ sensitivity. Furthermore, MSC-AS1 knockdown inhibited TMZ-resistant glioma growth in vivo by regulating miR-373-3p/CPEB4 axis through PI3K/Akt pathway. Collectively, MSC-AS1 knockdown suppressed cell growth and the chemoresistance of glioma cells to TMZ by regulating miR-373-3p/CPEB4 axis in vitro and in vivo through activating PI3K/Akt pathway.

Eplerenone Prevents Atrial Fibrosis via the TGF-β Signaling Pathway

Cardiology ◽  
2017 ◽  
Vol 138 (1) ◽  
pp. 55-62 ◽  
Author(s):  
Lili Du ◽  
Mu Qin ◽  
Yi Yi ◽  
Xiaoqing Chen ◽  
Weifeng Jiang ◽  
...  
Keyword(s):  
Protein Expression ◽  
Western Blotting ◽  
Feedback Regulation ◽  
Underlying Mechanism ◽  
In Vivo Studies ◽  
Atrial Fibrosis ◽  
Qrt Pcr ◽  
Tgf Β1

Objectives: Eplerenone (EPL), an antagonist of the mineralocorticoid receptor, is beneficial for atrial fibrillation and atrial fibrosis. However, the underlying mechanism remains less well known. We aimed to investigate the effect of EPL on atrial fibrosis using a mouse with selective atrial fibrosis and to explore the underlying mechanisms. Methods: EPL-treated MHC-TGFcys33ser transgenic mice that have selective atrial fibrosis (Tx+EPL mice), as well as control mice, were used for in vivo studies including histological analyses, Western blotting, and qRT-PCR studies. TGF-β1-stimulated atrial fibroblasts were treated with EPL or vehicle for the in vitro studies including Western blotting and qRT-PCR studies. In addition, Smad7 siRNA was used to knock down Smad7. Results: EPL inhibited atrial fibrosis in the Tx mice. In addition, EPL suppressed the expression of fibrosis-related molecules induced by TGF-β1 in vivo and in vitro. This occurred in concert with a downregulation of Smad7 protein expression and an upregulation of p-Smad2/3 protein expression. In addition, knockdown of Smad7 by siRNA abolished the protective roles of EPL. Conclusions: EPL inhibited atrial fibrosis in Tx mice. The underlying mechanism may involve increased protein expression of Smad7, which enhances the inhibitory feedback regulation of TGF-β1/Smad signaling.

scholarly journals Sp1-activated Long Noncoding RNA NCK1-AS1 Facilitates Cell Growth of Breast Cancer via Sponging miR-361-5p 

2020 ◽  
Author(s):  
Hongyao Jia ◽  
Di Wu ◽  
Zhiru Zhang ◽  
Sijie Li
Keyword(s):  
Breast Cancer ◽  
Cell Growth ◽  
Noncoding Rna ◽  
Malignant Tumors ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Cancer Tissues ◽  
Sphere Formation ◽  
First Time

Abstract Background: Breast cancer (BC) is one of the most lethal and malignant tumors in the world. Accumulating documents have illuminated the vital roles of long noncoding RNAs (lncRNAs) which are closely related to the progression of human cancers, including BC. LncRNA NCK1-AS1 was reported to be an oncogene in some cancers; nevertheless, its functionality is not well elucidated in BC. Methods: NCK1-AS1 expression status was detected in human breast cancer tissues and cell lines by the means of RT-qPCR. The impacts of NCK1-AS1 deficiency on breast cancer cell stemness, viability, proliferation, migration, invasion, and apoptosis were measured in vitro via sphere formation assay, western blot analysis, CCK-8, colony formation, transwell, TUNEL and flow cytometry analysis. Furthermore, luciferase reporter assay, RIP and ChIP were used to verify the mutual effects between molecules.Results: In this study, lncRNA NCK1-AS1 was verified to be elevated in BC tissues and cells. In addition, NCK1-AS1 silencing hampered BC cell growth by inhibiting cell stemness, viability, proliferation, migration and invasion as well as promoting cell apoptosis. Moreover, NCK1-AS1 functioned as a molecular sponge for miR-361-5p and modulated SP1 expression. Rescue experiments signified that SP1 overexpression restored NCK1-AS1 silencing-mediated suppression on BC cell growth. Importantly, SP1 was uncovered to bind with NCK1-AS1 promoter, suggesting a positive feedback loop of NCK1-AS1/miR-361-5p/SP1 in BC cells. Conclusion: Taken together, these findings showed for the first time that NCK1-AS1 served as a carcinogenic gene in BC cells via combining with miR-361-5p to upregulate SP1.

scholarly journals MiRNA-26a Contributes to the Acquisition of Malignant Behaviors of Doctaxel-Resistant Lung Adenocarcinoma Cells through Targeting EZH2

2017 ◽  
Vol 41 (2) ◽  
pp. 583-597 ◽  
Author(s):  
Jing Chen ◽  
Yuejuan Xu ◽  
Leilei Tao ◽  
Yan Pan ◽  
Kai Zhang ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Western Blotting ◽  
Human Cancer ◽  
Flow Cytometric Analysis ◽  
Human Lung Cancer ◽  
Luciferase Reporter ◽  
Apoptosis Rate ◽  
Qrt Pcr

Background/Aims: Accumulating evidence revealed that microRNAs (miRNAs) have been demonstrated as critical molecules in tumor development and progression. MiR-26a, located in a fragile chromosomal region associated with various human cancer, has been reported to be involved in regulating various cellular process, such as proliferation, apoptosis and invasion through targeting multiple oncogene. Docetaxel-mediated chemotherapy has been applied in improving the survival and prognosis of patients with advanced lung adenocarcinoma (LAD). However, chemoresistance remains a major impediment to clinical application of this agent. It has been presented that decreased miR-26a expression lead to cisplatin resistance and promoted growth and migration in human lung cancer. Enhancer of zeste homolog 2 (EZH2) is the target of miR-26a. The present study aimed to investigate the function of miR-26a/EZH2 in the acquisition of malignant behaviors of LAD. Methods: MiR-26a and EZH2 expression levels in the dcetaxel-insensitive groups (n = 19) and the docetaxel-sensitive groups (n = 18) were assessed by qRT-PCR. Colony formation assay, flow cytometric analysis, wound healing assay, cell transwell assays and western blotting were performed to assess the effects of miR-26a on proliferation, apoptosis and epithelial-to-mesenchymal (EMT) phenotypes in docetaxel resistant LAD cells in vitro. Xenograft transplantation, immunohistochemistry, tunel assays and western blotting assays were employed to demonstrate the role of miR-26a in docetaxel resistant LAD cells in vivo. The expression level of EZH2 in docetaxel-resistant LAD cells and corresponding parental cells was detected by qRT-PCR and western blotting. The relationship between miR-26a and EZH2 was confirmed by luciferase reporter assay. And rescue assays were performed to further confirm that miRNA-26a contributes to the acquisition of malignant behaviors of docetaxel-resistant LAD cells through targeting EZH2. Results: MiR-26a was significantly down-regulated in the dcetaxel-insensitive groups (n = 19) compared with the docetaxel-sensitive groups (n = 18) assessed by qRT-PCR. MiR-26a decreased the proliferation, increased the apoptosis rate and reversed EMT to MET of docetaxel-resistant LAD cells both in vivo and vitro. EZH2 was confirmed as target of miR-26a. Rescue assays further verified that the function of miR-26a exerts in docetaxel-resistant LAD cells is through targeting EZH2. Conclusions: Our data revealed that overexpression of miR-26a in docetaxel-resistant LAD cells could decrease the proliferation, increase the apoptosis rate and reverse EMT to MET of docetaxel-resistant LAD cells both in vivo and vitro and such function is partially exerted via downregulating EZH2. MiR-26a/EZH2 signal pathway makes contribute to the malignant phenotype of docetaxel-resistant of LAD cells which indicated that miR-26a exerts pivotal functions in the molecular etiology of chemoresistant lung adenocarcinoma.

scholarly journals LncRNA HAS2-AS1 Promotes Glioblastoma Proliferation by Sponging miR-137

2021 ◽  
Vol 11 ◽  
Author(s):  
Yalin Lu ◽  
Gaochao Guo ◽  
Rujun Hong ◽  
Xingjie Chen ◽  
Yan Sun ◽  
...  
Keyword(s):  
Underlying Mechanism ◽  
Cell Counting ◽  
Luciferase Reporter ◽  
Tumor Type ◽  
Downstream Target ◽  
Qrt Pcr ◽  
Lysine Specific Demethylase ◽  
Reporter Assays

GBM (Glioblastoma multiform) is the most malignant tumor type of the central nervous system and has poor diagnostic and clinical outcomes. LncRNAs (Long non-coding RNAs) have been reported to participate in multiple biological and pathological processes, but their underlying mechanism remains poorly understood. Here, we aimed to explore the role of the lncRNA HAS2-AS1 (HAS2 antisense RNA 1) in GBM. GSE103227 was analyzed, and qRT-PCR was performed to measure the expression of HAS2-AS1 in GBM. FISH (Fluorescence in situ hybridization) was performed to verify the localization of HAS2-AS1. The interaction between HAS2-AS1 and miR-137 (microRNA-137) was predicted by LncBook and miRcode followed by dual‐luciferase reporter assays, and the relationships among HAS2-AS1, miR-137 and LSD1 (lysine-specific demethylase 1) were assessed by WB (western blot) and qRT-PCR. Colony formation and CCK-8 (cell counting kit-8) assays were performed as functional tests. In vivo, nude mice were used to confirm the function of HAS2-AS1. HAS2-AS1 expression was upregulated in GBM cell lines, and HAS2-AS1 was localized mainly in the cytoplasm. In vitro, high HAS2-AS1 expression promoted proliferation, and knockdown of HAS2-AS1 significantly inhibited proliferation. Furthermore, HAS2-AS1 functioned as a ceRNA (competing endogenous RNA) of miR-137, leading to the disinhibition of its downstream target LSD1. The miR-137 level was downregulated by HAS2-AS1 overexpression and upregulated by HAS2-AS1 knockdown. In a subsequent study, LSD1 expression was negatively regulated by miR-137, while miR-137 reversed the LSD1 expression levels caused by HAS2-AS1. These results were further supported by the nude mouse tumorigenesis experiment; compared with xenografts with high HAS2-AS1 expression, the group with low levels of HAS2-AS1 exhibited suppressed proliferation and better survival. We conclude that lncRNA HAS2-AS1 promotes proliferation by functioning as a miR‐137 decoy to increase LSD1 levels and thus might be a possible biomarker for GBM.

scholarly journals Linc00460/miR-641 promoted promotes proliferation and autophagy of breast cancer

2020 ◽  
Author(s):  
Wei Zhang ◽  
Huimin Zhang ◽  
Bin Wang ◽  
Yu Ren ◽  
Jianjun He ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Luciferase Reporter ◽  
In Vitro Tests ◽  
Luciferase Reporter Gene ◽  
Qrt Pcr ◽  
In Vivo Tests ◽  
Worse Prognosis

Abstract This manuscript aimed to investigate the oncogenic role of linc00460 in breast cancer both in vivo and in vitro and then specify its downstream microRNAs to build a ceRNA network. As for in vivo tests, we used subgroup analysis and nomogram for survival analysis. As for in vitro tests, qRT-PCR, CCK-8 and Dual luciferase reporter gene were used. Linc00460 expressed highly in breast cancer. The nomogram indicated that linc00460 was associated with worse prognosis. Linc00460 might negatively regulate miR-641 to promote the proliferation and autophagy of breast cancer cell. In conclusion, linc00460 could be a risk factor for breast cancer by regulating miR-641; and it has the potential to be a novel biomarker.

scholarly journals Estrogenicity of essential oils is not required to relieve symptoms of urogenital atrophy in breast cancer survivors

2018 ◽  
Vol 10 ◽  
pp. 175883591876618 ◽  
Author(s):  
Bruno M. Simões ◽  
Bertram Kohler ◽  
Robert B. Clarke ◽  
Jacqui Stringer ◽  
Lily Novak-Frazer ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Antifungal Activity ◽  
Cell Growth ◽  
Cancer Survivors ◽  
Essential Oils ◽  
Breast Cancer Survivors ◽  
Luciferase Reporter ◽  
Systemic Absorption ◽  
Urogenital Atrophy

Background Urogenital atrophy (UA) is a common treatment-limiting side effect of endocrine therapies. Topical estrogen is effective but systemic absorption may counter aromatase inhibitor efficacy. Numerous complementary approaches are marketed for use in UA without rigorous testing of their estrogenicity. We tested multiple essential oils in cancer cell growth and estrogen reporter assays in vitro and assessed clinical outcomes with the essential oil pessaries (EOPs) in breast cancer survivors with UA. Methods Effects on cell growth were tested in hormone-dependent (MCF-7) and -independent (MDA-MB-231) cell lines using the sulforhodamine-B assay. An estrogen response element (ERE) luciferase reporter assay was used to assess estrogenicity directly. Antifungal activity against two common pathogenic yeasts was assessed using standard microdilution methods. EOPs were offered to breast cancer survivors with symptomatic UA and the service evaluated using serial questionnaires. Results Two essential oils, Cymbopogon martinii and Pelargonium graveolens, demonstrated marked estrogenicity, stimulating ER+ cell growth and ERE-luciferase reporter activity to levels seen with premenopausal estradiol concentrations. Additional oils were screened for estrogenicity and Lavandula angustifolia and Chamaemelum nobile identified as non/minimally estrogenic. The antifungal activity of this combination of oils was confirmed. A second cohort of breast cancer survivors with UA received the second generation EOP with comparable improvement in symptom scores suggesting that estrogenicity may not be required for optimal therapy of UA. Conclusion Certain essential oils demonstrate profound estrogenicity and caution should be exercised before their use in breast cancer survivors. Our minimally estrogenic pessary will be formally tested in clinical trials.

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