MicroRNA-451 regulates chemoresistance in renal cell carcinoma by targeting ATF-2 gene

Renal cell carcinoma (RCC) is a malignant tumor, which severely threatens human’s life, moreover, the multi-drug resistance (MDR) under RCC undoubtedly strengthen the difficulties in the treatment. MiR-451 has been considered to play an important role in regulation of MDR in several cancers, but the role of it in MDR of RCC has not been explored. This study aims to explore the mechanism of miR-451 as a target to regulate chemotherapy resistance, which is crucial for further exploring novel therapy for RCC. Two human cell lines (ACHN and GRC-1) were performed in this study and adriamycin (ADM) was used to construct MDR cell lines. qRT-PCR was used to determine the mRNA expression of miR-451 and ATF-2. Weston blot was used to determine protein expression. MTT assay and flow cytometry were used for assessing cell viability and apoptosis, individually. Luciferase reporter assay was used to detect the targeting of miR-451 and ATF-2. Results presented that the expression of miR-451 was higher in low MDR cell line (ACHN) comparing with the high MDR cell line (GRC-1), while the expression of ATF-2 revealed an opposite results. MiR-451 targeted ATF-2 and regulated its expression. Overexpression of miR-451 strengthened drug resistance, decreased cell viability, and increased cell apoptosis of GRC-1 pretreated by ADM, while overexpressed ATF-2 reversed the effect induced by miR-451 overexpression. Then miR-451 knockdown improved drug susceptibility, decreased cell apoptosis, and increased cell viability of ACHN induced by ADM, however, ATF-2 suppression reversed the low rate of cell apoptosis and high rate of cell viability induced by miR-451 knockdown. Our results revealed that miR-451 regulates the drug resistance of RCC by targeting ATF-2 gene, which might be critical for overcoming MDR in RCC patients. Impact statement This is the first study to emphasize the expression of miR-451 on regulating multi-drug resistance (MDR) in renal cell carcinoma (RCC). Our study found that miR-451 regulates the drug resistance of RCC by targeting ATF-2, which might be critical for overcoming MDR in RCC patients. This study not only provides solid theory foundation for the clinical therapy, but also offers unique insights for the further RCC research. Furthermore, the study helps us to understand the mechanism of MDR, which was crucial for identifying the chemoresistance on several related tumors.

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Metabolomic Analysis to Elucidate Mechanisms of Sunitinib Resistance in Renal Cell Carcinoma

Metabolites ◽

10.3390/metabo11010001 ◽

2020 ◽

Vol 11 (1) ◽

pp. 1

Author(s):

Tomonori Sato ◽

Yoshihide Kawasaki ◽

Masamitsu Maekawa ◽

Shinya Takasaki ◽

Kento Morozumi ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Energy Metabolism ◽

Cell Carcinoma ◽

Cell Line ◽

Cell Lines ◽

Cancer Progression ◽

Renal Cell ◽

Treatment Resistance ◽

Tca Cycle ◽

Glutamine Uptake

Metabolomics analysis possibly identifies new therapeutic targets in treatment resistance by measuring changes in metabolites accompanying cancer progression. We previously conducted a global metabolomics (G-Met) study of renal cell carcinoma (RCC) and identified metabolites that may be involved in sunitinib resistance in RCC. Here, we aimed to elucidate possible mechanisms of sunitinib resistance in RCC through intracellular metabolites. We established sunitinib-resistant and control RCC cell lines from tumor tissues of RCC cell (786-O)-injected mice. We also quantified characteristic metabolites identified in our G-Met study to compare intracellular metabolism between the two cell lines using liquid chromatography-mass spectrometry. The established sunitinib-resistant RCC cell line demonstrated significantly desuppressed protein kinase B (Akt) and mesenchymal-to-epithelial transition (MET) phosphorylation compared with the control RCC cell line under sunitinib exposure. Among identified metabolites, glutamine, glutamic acid, and α-KG (involved in glutamine uptake into the tricarboxylic acid (TCA) cycle for energy metabolism); fructose 6-phosphate, D-sedoheptulose 7-phosphate, and glucose 1-phosphate (involved in increased glycolysis and its intermediate metabolites); and glutathione and myoinositol (antioxidant effects) were significantly increased in the sunitinib-resistant RCC cell line. Particularly, glutamine transporter (SLC1A5) expression was significantly increased in sunitinib-resistant RCC cells compared with control cells. In this study, we demonstrated energy metabolism with glutamine uptake and glycolysis upregulation, as well as antioxidant activity, was also associated with sunitinib resistance in RCC cells.

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Inhibition of SMYD2 suppresses tumor progression by down-regulating microRNA-125b and attenuates multi-drug resistance in renal cell carcinoma: Erratum

Theranostics ◽

10.7150/thno.68810 ◽

2022 ◽

Vol 12 (1) ◽

pp. 419-420

Author(s):

Libin Yan ◽

Beichen Ding ◽

Haoran Liu ◽

Yangjun Zhang ◽

Jin Zeng ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Drug Resistance ◽

Tumor Progression ◽

Cell Carcinoma ◽

Renal Cell ◽

Multi Drug Resistance

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Inhibition of SMYD2 suppresses tumor progression by down-regulating microRNA-125b and attenuates multi-drug resistance in renal cell carcinoma

Theranostics ◽

10.7150/thno.37628 ◽

2019 ◽

Vol 9 (26) ◽

pp. 8377-8391 ◽

Cited By ~ 3

Author(s):

Libin Yan ◽

Beichen Ding ◽

Haoran Liu ◽

Yangjun Zhang ◽

Jin Zeng ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Drug Resistance ◽

Tumor Progression ◽

Cell Carcinoma ◽

Renal Cell ◽

Multi Drug Resistance

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MiR-125b promotes migration and invasion by targeting vitamin D receptor in renal cell carcinoma

10.21203/rs.3.rs-19608/v1 ◽

2020 ◽

Author(s):

XiYuan He ◽

ShangFan Liao ◽

DongMing Lu ◽

FaBiao Zhang ◽

yongyang wu

Keyword(s):

Renal Cell Carcinoma ◽

Vitamin D ◽

Cell Carcinoma ◽

Cell Lines ◽

Vitamin D Receptor ◽

Cell Apoptosis ◽

Renal Cell ◽

Migration And Invasion ◽

Proliferation And Metastasis ◽

And Behavior

Abstract Background To investigate the expression of miR-125b and vitamin D receptor (VDR) in renal cell carcinoma (RCC) and assess the possible association between them. Then, to elucidate whether miR-125b can regulate the expression of VDR and affect proliferation and metastasis in RCC. Methods The expression of miR-125b was detected by quantitative real-time polymerase chain reaction (RT-PCR) in RCC cell lines. MiR-125b mimic and inhibitor were employed to measure the function and behavior of miR-125b in RCC cell lines. The relationship between miR-125 and VDR was verified using luciferase assays, and their expression was also examined in primary tumor and normal peritumoral kidney tissues in 20 clear cell RCC (ccRCC) samples. Results Overexpression of miR-125b promoted migration and invasion and reduced cell apoptosis in ACHN cells, while inhibition of miR-125b suppressed migration and invasion and induced cell apoptosis in 786-O cells. Overexpression of miR-125b decreased VDR expression via targeting VDR. Expression of miR-125b mRNA was significantly higher in ccRCC tissues than in normal adjacent tissues, and the expression of miR-125b mRNA negatively correlated with that of VDR (r=-0.444, p=0.04). Conclusion Overexpression of miR-125b decreased the expression of VDR and the promoted migration and invasion of RCC cells; in addition, there was a negative correlation between miR-125b and VDR expression in ccRCC.

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Inhibition of SMYD2 suppresses tumor progression by down-regulating microRNA-125b and attenuates multi-drug resistance in renal cell carcinoma

European Urology Open Science ◽

10.1016/s2666-1683(20)33579-5 ◽

2020 ◽

Vol 19 ◽

pp. e1481

Author(s):

L. Yan ◽

H. Liu ◽

G. Sun ◽

Y. Zhang

Keyword(s):

Renal Cell Carcinoma ◽

Drug Resistance ◽

Tumor Progression ◽

Cell Carcinoma ◽

Renal Cell ◽

Multi Drug Resistance

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Ovatodiolide Targetsβ-Catenin Signaling in Suppressing Tumorigenesis and Overcoming Drug Resistance in Renal Cell Carcinoma

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2013/161628 ◽

2013 ◽

Vol 2013 ◽

pp. 1-16 ◽

Cited By ~ 12

Author(s):

Jar-Yi Ho ◽

Ren-Jun Hsu ◽

Chieh-Lin Wu ◽

Wen-Liang Chang ◽

Tai-Lung Cha ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Drug Resistance ◽

Cell Carcinoma ◽

Cell Lines ◽

Renal Cell ◽

Synergistic Effects ◽

Biological Effects ◽

Mouse Xenograft Model ◽

Signaling Inhibitors

Dysregulatedβ-catenin signaling is intricately involved in renal cell carcinoma (RCC) carcinogenesis and progression. Determining potentialβ-catenin signaling inhibitors would be helpful in ameliorating drug resistance in advanced or metastatic RCC. Screening forβ-catenin signaling inhibitors involvedin silicoinquiry of the PubChem Bioactivity database followed by TCF/LEF reporter assay. The biological effects of ovatodiolide were evaluated in 4 RCC cell linesin vitroand 2 RCC cell lines in a mouse xenograft model. The synergistic effects of ovatodiolide and sorafenib or sunitinib were examined in 2 TKI-resistant RCC cell lines. Ovatodiolide, a pure compound ofAnisomeles indica, inhibitedβ-catenin signaling and reduced RCC cell viability, survival, migration/invasion, andin vitrocell orin vivomouse tumorigenicity. Cytotoxicity was significantly reduced in a normal kidney epithelial cell line with the treatment. Ovatodiolide reduced phosphorylatedβ-catenin (S552) that inhibitedβ-catenin nuclear translocation. Moreover, ovatodiolide decreasedβ-catenin stability and impaired the association ofβ-catenin and transcription factor 4. Ovatodiolide combined with sorafenib or sunitinib overcame drug resistance in TKI-resistant RCC cells. Ovatodiolide may be a potentβ-catenin signaling inhibitor, with synergistic effects with sorafenib or sunitinib, and therefore, a useful candidate for improving RCC therapy.

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Hydroxychloroquine Potentiates Apoptosis Induced by PPARα Antagonist in 786-O Clear Cell Renal Cell Carcinoma Cells Associated with Inhibiting Autophagy

PPAR Research ◽

10.1155/2021/6631605 ◽

2021 ◽

Vol 2021 ◽

pp. 1-14

Author(s):

Ruizhe Mao ◽

Jian Shi ◽

Xuyi Ma ◽

Haiyan Xu

Keyword(s):

Renal Cell Carcinoma ◽

Cell Viability ◽

Cell Carcinoma ◽

Cell Line ◽

Renal Cell ◽

Clear Cell ◽

Therapeutic Drug ◽

Peroxisome Proliferator ◽

Cell Renal Cell Carcinoma ◽

Ccrcc Cell

Clear cell renal cell carcinoma (ccRCC) is the major pathological pattern of renal cell carcinoma. The ccRCC cells exhibit a certain degree of inherent drug resistance due to some genetic mutations. In recent years, peroxisome proliferator-activated receptor-α (PPARα) antagonists have been reported as a targeted therapeutic drug capable of inducing apoptosis and cell cycle arrest in the ccRCC cell line. Autophagy, which can be induced by stress in eukaryotic cells, plays a complex role in the proliferation, survival, and death of tumor cells. In our study, we found that the expression of PPARα was low in highly differentiated ccRCC tissues and 786-O cell line but high in poorly differentiated ccRCC tissues. The level of PPARα expression in ccRCC tissues is correlated to the grade of differentiation, but not to the sex or age of ccRCC patients. The findings also revealed that the PPARα antagonist GW6471 can lower cell viability and induce autophagy in the 786-O ccRCC cell line. This autophagy can be inhibited by hydroxychloroquine. When treated with a combination of hydroxychloroquine and GW6471, the viability of the 786-O cells was decreased further when compared to the treatment with GW6471 or hydroxychloroquine alone, and apoptosis was promoted. Meanwhile, when human kidney 2 cells were cotreated with hydroxychloroquine and GW6471, cell viability was only slightly influenced. Hence, our finding indicates that the combination of GW6471 and hydroxychloroquine may constitute a novel and potentially effective treatment for ccRCC. Furthermore, this approach is likely to be safe owing to its minimal effects on normal renal tissues.

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SNHG16 promotes cell proliferation and inhibits cell apoptosis via regulation of the miR-1303p/STARD9 axis in renal cell carcinoma

10.1101/2021.03.10.434814 ◽

2021 ◽

Author(s):

Tao Cheng ◽

Weibing Shuang ◽

Dawen Ye ◽

Wenzhi Zhang ◽

Zhao Yang ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Cell Proliferation ◽

Cell Carcinoma ◽

Cell Apoptosis ◽

Renal Cell ◽

Target Gene ◽

Quantitative Polymerase Chain Reaction ◽

Cell Counting ◽

Luciferase Reporter ◽

High Morbidity

AbstractBackgroundRenal cell carcinoma (RCC) is a fatal malignant tumor with high morbidity. Numerous medical studies have suggested that long noncoding RNAs (lncRNAs) exert their biological function on various cancerous progresses. Herein, functions of lncRNA SNHG16 in RCC cells and the mechanism medicated by SNHG16 were investigated.MethodsThe expression levels of SNHG16 and its downstream genes in RCC cells and tissues were examined utilizing reverse transcription quantitative polymerase chain reaction analyses. Cell counting kit-8 and 5-Ethynyl-2’-deoxyuridine assays were carried out to evaluate the proliferation of RCC cells, and flow cytometry analyses were employed to determine the apoptosis of RCC cells. Western blot analysis was applied to examine protein levels associated with cell proliferation and apoptosis. The combination between SNHG16 and miRNA as well as miRNA and its target gene were explored by luciferase reporter, RNA pull down, and RNA immunoprecipitation assays.ResultsThe significant upregulation of SNHG16 was observed in RCC tissues and cells. SNHG16 downregulation inhibited the proliferation and promoted the apoptosis of RCC cells. In addition, SNHG16 served as a competing endogenous RNA for miR-1301-3p, and STARD9 was a target gene of miR-1301-3p in RCC cells. SNHG16 upregulated STARD9 expression by binding with miR-1301-3p in RCC cells. Rescue assays validated that SNHG16 promoted RCC cell promotion and induced RCC cell apoptosis by upregulating STARD9 expression.ConclusionsSNHG16 promotes RCC cell proliferation and suppresses RCC cell apoptosis via interaction with miR-1301-3p to upregulate STARD9 expression in RCC cells.

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Identification of an appropriate cell line for the renal cell carcinoma model

10.21203/rs.3.rs-39971/v1 ◽

2020 ◽

Author(s):

In Youb Chang ◽

Takbum Ohn ◽

Daeun Moon ◽

Young Hee Maeng ◽

Bo Gun Jang ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Cell Carcinoma ◽

Cell Line ◽

Cell Lines ◽

Renal Cell ◽

Tissue Microarrays ◽

Future Research ◽

Proximal Tubule Cells

Abstract Background Although renal cell carcinoma (RCC) is known to be susceptible to ferroptosis, we found primary RCC cells showed resistance to ferroptosis and aimed to investigate a feasible candidate for an appropriate cell line for the RCC model. Methods Glutathione peroxidase 4 (GPX4) immunostaining was adopted in the RCC tissue microarrays. Normal human proximal tubule cells (HK-2) and RCC cell lines were used for the MTT assay, Western blotting, sphere-forming assay, and orthotopic injection of athymic Balb/c-nude mice. Results GPX4 immunostaining showed low intensity compared to the normal kidney, which coincided with the ferroptosis-susceptibility of RCC. Primary RCC cell lines (Caki-2, SNU-333, SNU-349, and SNU-1272) showed resistance to 5-fluorouracil and a GPX4 inhibitor compared to the HK-2 cells and to metastatic RCC cells (Caki-1). The Caki-2 cells showed increased GPX4 and xCT, and the SNU-333 cells showed increased ferritin heavy chain (FTH1) compared to the other RCC cells. The Caki-2 cells showed increased aSMA, fibronectin, vimentin, and SNAIL, and the SNU-333 cells showed increased aSMA, E-cadherin, and EpCAM. The Caki-2 cells showed increased Sox-2 and CD105, and the SNU-333 cells showed increased c-Myc and Lgr5. The Caki-1 and SNU-333 cells formed spheres in vitro and orthotopic RCC masses in vivo. The injected SNU-333 tumor only showed high intensities of CD10 and PAX8, consistent with the diagnostic criteria for RCC. Conclusions The primary RCC cell lines used in this study were more resistant to ferroptosis and 5-fluorouracil, and the SNU-333 cells showed tumor-initiating capacities in vitro and in vivo. These results suggest that SNU-333 might be suitable as a orthotopic RCC model for future research.

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