Knockdown of TRIM66 Suppresses the Proliferation, Migration, Invasion and Glycolysis in Cholangiocarcinoma Cells

Abstract Background: The tripartite motif (TRIM) family proteins feature highly conserved order of domains in the RBCC motif and most of them play an essential role in various cellular processes. Recently, increasing evidence has shown association of TRIM proteins with cancer development. In this study, we examined the expression pattern and biological functions of TRIM66 in cholangiocarcinoma (CCA).Methods: Western blot was performed for the protein levels of TRIM66, E-cadherin, α-catenin, N-cadherin, vimentin, p-PI3K, PI3K, p-Akt and Akt. MTT assay, wound healing assay and transwell assay were conducted for cell proliferation, migration and invasion, respectively. Glucose uptake and lactate production were determined using specific kits.Results: TRIM66 was overexpressed in CCA tissues and cell lines. In addition, knockdown of TRIM66 significantly inhibited proliferation, migration, invasion and glycolysis of CCA cells. Moreover, TRIM66 silencing obviously decreased levels of phosphorylated PI3K and Akt in CCA cells.Conclusion: Our study provided a novel insight into the roles of TRIM66 in CCA and suggested TRIM66 as a promising therapeutic target for CCA treatment.

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Downregulation of KIF21B by miR-132-3p suppresses cellular functions in gastric cancer via regulating Wnt/β-catenin signaling pathway

10.21203/rs.3.rs-850811/v1 ◽

2021 ◽

Author(s):

Bingtian Liu ◽

Ling Qiang ◽

Bingxin Guan ◽

Zhipeng Ji

Keyword(s):

Gastric Cancer ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Small Cell Lung ◽

Cellular Functions ◽

Transwell Assay ◽

Microarray Gene Expression ◽

Protein Levels ◽

Normal Tissues ◽

Kinesin Family

Abstract Recently, kinesin family member 21B (KIF21B) has been reported to be an oncogene in non-small cell lung cancer and hepatocellular carcinoma. However, the functional role and related mechanisms underlying gastric cancer (GC) pathogenesis remain largely uncovered. Here, we first found that the expression of KIF21B was upregulated in GC tissues compared with adjunct normal tissues by analysis of Oncomine microarray gene expression datasets and clinical specimens. Knockdown of KIF21B significantly suppressed the proliferation, migration and invasion in GC cell lines using CCK-8 assay and transwell assay. By luciferase reporter assay, KIF21B was confirmed as the target of miR-132-3p in GC cells and suppressed after miR-132-3p overexpression. Moreover, miR-132-3p was down-regulated and inversely correlated with KIF21B expression in GC tissues. Further functional experiments demonstrated that overexpression of KIF21B remarkedly reversed the suppressive effects of miR-132-3p overexpression on GC cell proliferation, migration and invasion. Furthermore, western blot analysis manifested that miR-132-3p overexpression downregulated the protein levels of Wnt1, c-Myc, β-catenin, PCNA and N-cadherin, and upregulated E-cadherin expression in GC cells, which were all alleviated after KIF21B overexpression. In summary, our findings provide the first evidence that down-regulation of KIF21B by miR-132-3p suppresses cellular functions in gastric cancer via regulating Wnt/β-catenin signaling.

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miR-577 suppressed the metastasis, EMT and viability via NF-κB pathway by targeting CXCL5 through in hepatocellular carcinoma

10.21203/rs.3.rs-27744/v1 ◽

2020 ◽

Author(s):

Lirui Tu ◽

Jing Liu ◽

Wei Li ◽

Xiuguang Song ◽

Hongwei Xu

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Viability ◽

Cell Invasion ◽

Great Role ◽

Transwell Assay ◽

Protein Levels ◽

Xenograft Growth ◽

Kaplan Meier ◽

Insight Into

Abstract Background Hepatocellular carcinoma (HCC) is a common malignant cancer worldwide. miR-577 have a role in inhibiting cell viability, metastasis in many tumors. This research was to explore the great role of miR-577 in hepatocellular carcinoma. Methods RT-qPCR and western blot were performed to evaluate the the miR-577 and genes mRNA and protein levels. Transwell assay and CCK-8 were applied to measure the viable and invasive abilities. Meanwhile, Kaplan-Meier method was used to assess the survival of HCC patients. Results miR-577 was downregulated in HCC tissues, which predicted a worse overall survival in HCC. miR-577 targeted to CXCL5 and mediated its expression in HCC. miR-577 suppressed cell invasion and EMT in HuH-7 cells. miR-577 inhibited cell viability via NF-κB pathway. In addition, miR-577 overxpression impaired the xenograft growth of HuH-7 cells. Conclusion miR-577 inhibited cell invasion, EMT and viability via NF-κB pathway by targeting to CXCL5 in HCC. The newly identified miR-577/CXCL5 axis provides novel insight into the pathogenesis of hepatocellular carcinoma.

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TRIM Proteins in Colorectal Cancer: TRIM8 as a Promising Therapeutic Target in Chemo Resistance

Biomedicines ◽

10.3390/biomedicines9030241 ◽

2021 ◽

Vol 9 (3) ◽

pp. 241

Author(s):

Flaviana Marzano ◽

Mariano Francesco Caratozzolo ◽

Graziano Pesole ◽

Elisabetta Sbisà ◽

Apollonia Tullo

Keyword(s):

Colorectal Cancer ◽

Clinical Practice ◽

Therapeutic Target ◽

Malignant Tumors ◽

Premature Death ◽

Chemotherapy Response ◽

Protein Levels ◽

Promising Therapeutic Target ◽

Trim Proteins

Colorectal cancer (CRC) represents one of the most widespread forms of cancer in the population and, as all malignant tumors, often develops resistance to chemotherapies with consequent tumor growth and spreading leading to the patient’s premature death. For this reason, a great challenge is to identify new therapeutic targets, able to restore the drugs sensitivity of cancer cells. In this review, we discuss the role of TRIpartite Motifs (TRIM) proteins in cancers and in CRC chemoresistance, focusing on the tumor-suppressor role of TRIM8 protein in the reactivation of the CRC cells sensitivity to drugs currently used in the clinical practice. Since the restoration of TRIM8 protein levels in CRC cells recovers chemotherapy response, it may represent a new promising therapeutic target in the treatment of CRC.

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Tumor Cell-Derived Exosomal hsa_circ_0017252 Suppresses the Tumorigenesis of Gastric Cancer Through Suppressing M2 Polarization of Macrophage

10.21203/rs.3.rs-907938/v1 ◽

2021 ◽

Author(s):

Jin Song ◽

Xiaolong Xu ◽

Shasha He ◽

Ning Wang ◽

Yunjing Bai ◽

...

Keyword(s):

Gastric Cancer ◽

Migration And Invasion ◽

Nanoparticle Tracking Analysis ◽

Transwell Assay ◽

Gene Expressions ◽

Protein Levels ◽

M2 Polarization ◽

Transmission Electron ◽

And Migration

Abstract Background: Gastric cancer (GC) is a source of global cancer death. MiR-17-5p is reported to regulate the tumorigenesis of GC. Meanwhile, hsa_circ_0017252 is known to be upregulated in GC. However, the relation between hsa_circ_0017252 and miR-17-5p in GC remains unclear. Methods: Cell viability, migration and invasion were tested by CCK-8 and transwell assay, respectively. Gene expressions were detected by RT-qPCR, and the protein levels in cells or exosomes were tested by western blot. The efficiency of exosomes isolation was investigated by transmission electron microscope (TEM) and nanoparticle tracking analysis (NTA). Meanwhile, cell apoptosis was tested by flow cytometry. In vivo model was constructed to assess the function of MKN45 cells-derived exosomal hsa_circ_0017252 in GC. Results: Hsa_circ_0017252 was verified to be significantly downregulated in GC tissues. Hsa_circ_0017252 upregulation significantly decreased the viability and migration of GC cells, and hsa_circ_0017252 could bind with miR-17-5p. Additionally, exosomal hsa_circ_0017252 reversed the polarization of M2 macrophages, and the polarized macrophages decreased the GC cell invasion. Furthermore, exosomes with upregulated hsa_circ_0017252 suppressed the tumor growth of GC. Conclusion: Delivery of hsa_circ_0017252 by GC cells-derived exosomes inhibits the tumorigenesis of GC through reversing M2 polarization of macrophages. Thus, our finding might provide a new method for GC treatment.

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Quercetin inhibits the tumorigenesis of colorectal cancer cells through downregulation of hsa_circ_0006990

10.21203/rs.3.rs-1019635/v1 ◽

2021 ◽

Author(s):

Bin Chen ◽

Haijuan Xiao ◽

Linguangjin Wu ◽

Ting Wang ◽

Shuyun Wang ◽

...

Keyword(s):

Colorectal Cancer ◽

Cancer Cells ◽

Luciferase Assay ◽

Migration And Invasion ◽

Colony Formation Assay ◽

Transwell Assay ◽

Protein Levels ◽

Colorectal Cancer Cells ◽

New Strategies ◽

Malignant Behavior

Abstract Background This study was intended to investigate the function of Quercetin in chemoresistant colorectal cancer (CRC) cells. In addition, this research aimed to explore the mechanism by which Quercetin regulates the malignant behavior of CRC cells. Methods To induce THP-1 cells into M2 tumor-associated macrophages (M2-TAMs), THP-1 cells were stimulated by PMA and IL-4. MDC staining was used to investigate the autophagy in M2-TAMs. Meanwhile, cell proliferation was tested by colony formation assay. In addition, wound healing and transwell assay were performed to detect the cell migration and invasion, respectively. Dual luciferase assay was used to investigate the correlation between hsa_circ_0006990 and miR-132-3p/miR-532-3p. Furthermore, mRNA and protein levels were detected by RT-qPCR and western blot, respectively. Results Quercetin suppressed autophagy of M2-TAMs. In addition, M2-TAMs significantly inhibited the apoptosis and promoted the proliferation of CRC cells, while this phenomenon was reversed by Quercetin. Meanwhile, the expression of hsa_circ_0006990 in CRC cells was decreased by M2-TAMs, while Quercetin reversed this phenomenon. Furthermore, overexpression of hsa_circ_0006990 significantly reversed the anti-tumor effect of Quercetin on CRC. Conclusion Quercetin inhibited the tumorigenesis of colorectal cancer cells through downregulation of hsa_circ_0006990. Thus, our study might shed new lights on exploring the new strategies against CRC.

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Nesfatin-1/Nucleobindin-2 Is a Potent Prognostic Marker and Enhances Cell Proliferation, Migration, and Invasion in Bladder Cancer

Disease Markers ◽

10.1155/2018/4272064 ◽

2018 ◽

Vol 2018 ◽

pp. 1-9 ◽

Cited By ~ 3

Author(s):

Guang-Ming Liu ◽

Zi-Qiang Xu ◽

Hong-Shun Ma

Keyword(s):

Bladder Cancer ◽

Poor Prognosis ◽

High Expression ◽

Migration And Invasion ◽

Wound Healing Assay ◽

Transwell Assay ◽

T24 Cells ◽

Tumor Growth And Metastasis

In recent researches, high expression of nesfatin-1/nucleobindin-2 (NUCB2) is linked to poor prognosis in prostate and colon cancer due to the enhancement in proliferation, migration, and invasion. However, the role of nesfatin-1 in bladder cancer is not clear. In this study, we examined the expression of NUCB2 in bladder cancer using immunohistochemistry and observed that its high expression was associated with recurrence and metastasis. In addition, the transwell assay and wound healing assay showed that cell migration and invasion were decreased with NUCB2 knockdown in T24 and 5637 cells. In vivo, tumor growth and metastasis were inhibited with shRNA treatment in T24 cells. Those results showed that NUCB2 played an important role in bladder cancer and could be considered a potent prognostic factor in bladder cancer.

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miR-126a-3p induces proliferation, migration and invasion of trophoblast cells in pre-eclampsia-like rats by inhibiting A Disintegrin and Metalloprotease 9

Bioscience Reports ◽

10.1042/bsr20191271 ◽

2019 ◽

Vol 39 (12) ◽

Cited By ~ 1

Author(s):

Shenglong Zhao ◽

Jiandong Wang ◽

Zheng Cao ◽

Lei Gao ◽

Yuanyuan Zheng ◽

...

Keyword(s):

Cell Cycle ◽

Wound Healing ◽

Underlying Mechanism ◽

Migration And Invasion ◽

Cell Cycle Distribution ◽

Wound Healing Assay ◽

Trophoblast Cells ◽

Transwell Assay ◽

Blank Group

Abstract The present study aimed to investigate the underlying mechanism of miR-126a-3p in the proliferation, migration and invasion of trophoblast cells in pre-eclampsia-like rats by targeting A Disintegrin and Metalloprotease 9 (ADAM9). First, the interaction between miR-126a-3p and ADAM9 was confirmed via biochemical assays. Placental tissues and trophoblast cells were then obtained. RNA in situ hybridization was performed in order to detect miR-126a-3p expression in the placenta. Subsequently, a series of biological assays, including reverse transcription-quantitative PCR (RT-qPCR), Western blotting, MTT assay, apoptosis assay, cell cycle assay, wound healing assay and transwell assay were adopted in order to determine the cell proliferation, cell cycle distribution, apoptotic rate, and migration and invasion of trophoblast cells in each group. The results revealed that miR-126a-3p was down-regulated in the placenta of pre-eclampsia-like rats. In vivo experiments’ results indicated that miR-126a-3p could inhibit ADAM9 expression, and induce cyclin D1, Matrix metalloproteinase (MMP) 2 (MMP-2), MMP-9 expression. MTT, apoptosis and cell cycle assay results revealed that trophoblast cells transfected with miR-126a-3p mimic or si-ADAM9 exhibited higher proliferative activity and a lower apoptotic rate compared with the blank group (all P<0.05). The wound healing assay and transwell assay results confirmed that, compared with the blank group, the migration and invasion ability of trophoblast cells in the miR-126a-3p mimic group and small interfering RNA (siRNA)-ADAM9 group were significantly increased (all P<0.05). Conversely, miR-126a-3p inhibitor treatment revealed the opposite effect (all P<0.05). In conclusion, the present study demonstrated that miR-126a-3p could enhance proliferation, migration and invasion, but decrease the apoptosis rate of trophoblast cells in pre-eclampsia-like rats through targeting ADAM9.

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Knockdown of YAP/TAZ Inhibits the Migration and Invasion of Fibroblast Synovial Cells in Rheumatoid Arthritis by Regulating Autophagy

Journal of Immunology Research ◽

10.1155/2020/9510594 ◽

2020 ◽

Vol 2020 ◽

pp. 1-11

Author(s):

Wei Zhou ◽

Qin Shen ◽

Hui Wang ◽

Jie Yang ◽

Chen Zhang ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

Migration And Invasion ◽

Binding Motif ◽

Transcriptional Coactivator ◽

Wound Healing Assay ◽

Transwell Assay ◽

Chain Reaction ◽

Polymerase Chain ◽

Novel Targets ◽

Fibroblast Like Synoviocytes

The purpose of this study was to investigate the effect of knockdown of the yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) on the migration and invasion of the rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS) and to preliminarily elucidate the mechanisms between YAP/TAZ and autophagy in the migration and invasion of RA-FLS. RA-FLS stable knockdown of YAP or TAZ was successfully established by using lentiviral-mediated gene knockdown techniques. Wound healing assay and Transwell assay were used to evaluate the effect of knockdown of YAP or TAZ on the migration and invasion of RA-FLS. Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) and western blotting assays were performed to examine the expression of indicated genes. The results showed that YAP and TAZ were upregulated in RA-FLS, and knockdown of YAP or TAZ inhibited the migration and invasion, reduced the expression of N-cadherin and Vimentin, and increased the accumulation of E-cadherin and β-catenin in RA-FLS. Our results also demonstrated that knockdown of YAP or TAZ promoted autophagy which increased the accumulation of LC3B-II and ULK1 and decreased the amount of SQSTM1/p62 in RA-FLS. Furthermore, our data displayed that inhibition of autophagy either with 3-MA or CQ can partially reverse the decrease of migration and invasion induced by YAP and TAZ knockdown in RA-FLS. Our experiments preliminarily revealed that YAP/TAZ and autophagy play important roles in the migration and invasion of RA-FLS, which might provide novel targets for the treatment of RA.

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Ubiquitination and degradation of MGMT by TRIM72 increases the sensitivity of uveal melanoma cells to Dacarbazine treatment

Cancer Biomarkers ◽

10.3233/cbm-210345 ◽

2021 ◽

pp. 1-10

Author(s):

Xun Li ◽

Cong Yang ◽

Ning Luo ◽

Yunzhi Yang ◽

Yan Guo ◽

...

Keyword(s):

Uveal Melanoma ◽

Negative Correlation ◽

Antitumor Effect ◽

Melanoma Cells ◽

Methyl Transferase ◽

Protein Levels ◽

Promising Therapeutic Target ◽

Tripartite Motif ◽

Increased Sensitivity ◽

Trim Proteins

Uveal melanoma (UM) is the most common primary intraocular malignancy in adults with high metastasis rates. The O6-methylguanine DNA methyl transferase (MGMT) is involved in chemoresistance of Dacarbazine (DTIC) treatment. Our previous study found that the combination of oncolytic adenovirus H101 and DTIC in the treatment of UM cells shows a synergistic antitumor effect mainly though down-regulation of MGMT. MGMT knockdown by shRNAs increases the sensitivity of uveal melanoma cells to DTIC treatment. The protein hemostasis of MGMT is important for the antitumor effect of DTIC. Tripartite motif-containing protein 72 (TRIM72) belongs to the tripartite motif (TRIM) proteins family and was identified as a novel E3 ligase for MGMT, which interacts with and mediates the ubiquitination of MGMT. TRIM72 knockdown increases the protein levels of MGMT, while reduces the ubiquitination of MGMT. Further study indicated that MGMT is highly expressed in UM cells, and the protein levels of MGMT and TRIM72 shows a negative correlation. UM cells that ectopically expressing TRIM72 shows increased sensitivity to DTIC treatment, which is consistent with the antitumor affect exhibited by H101. These results suggest that TRIM72 is a promising therapeutic target for UM treatment.

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Advances in determining signaling mechanisms of ceramide and role in disease

The Journal of Lipid Research ◽

10.1194/jlr.s092874 ◽

2019 ◽

Vol 60 (5) ◽

pp. 913-918 ◽

Cited By ~ 16

Author(s):

Jeffrey L. Stith ◽

Fabiola N. Velazquez ◽

Lina M. Obeid

Keyword(s):

Effector Proteins ◽

Membrane Properties ◽

Biological Functions ◽

Cellular Processes ◽

The Past ◽

Signaling Mechanisms ◽

Metabolizing Enzyme ◽

Insight Into

Ceramide is a critical bioactive lipid involved in diverse cellular processes. It has been proposed to regulate cellular processes by influencing membrane properties and by directly interacting with effector proteins. Advances over the past decade have improved our understanding of ceramide as a bioactive lipid. Generation and characterization of ceramide-metabolizing enzyme KO mice, development of specific inhibitors and ceramide-specific antibodies, use of advanced microscopy and mass spectrometry, and design of synthetic ceramide derivatives have all provided insight into the signaling mechanisms of ceramide and its implications in disease. As a result, the role of ceramide in biological functions and disease are now better understood, with promise for development of therapeutic strategies to treat ceramide-regulated diseases.

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