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

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

10.21203/rs.3.rs-1126402/v1 ◽

2021 ◽

Author(s):

Lei Jiang ◽

Yuqiang Li ◽

Yan Li ◽

Tao Yang ◽

Dongsheng Li ◽

...

Keyword(s):

Lactate Production ◽

Migration And Invasion ◽

Biological Functions ◽

Wound Healing Assay ◽

Transwell Assay ◽

Protein Levels ◽

Cellular Processes ◽

Promising Therapeutic Target ◽

Trim Proteins ◽

Insight Into

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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Features of the lymphocytic microenvironment in metastatic uveal melanoma

Уральский медицинский журнал ◽

10.52420/2071-5943-2021-20-2-36-42 ◽

2021 ◽

Vol 20 (2) ◽

pp. 36-42

Author(s):

A. Y. Shamanova ◽

E. L. Kazachkov ◽

I. Е. Panova ◽

A. V. Vazhenin ◽

Т. N. Shamaeva ◽

...

Keyword(s):

Immune Response ◽

Uveal Melanoma ◽

Antitumor Effect ◽

Primary Melanoma ◽

Distant Metastases ◽

Melanoma Cells ◽

The Body ◽

Immune Reactivity ◽

Tumor Metastases ◽

Cd4 Lymphocytes

Introduction. Uveal melanoma is a malignant neoplasm of the vascular tract of the eye. Prevention of metastasis of this tumor is one of the main tasks in order to increase the rates of relapse-free survival of patients. Despite the pronounced immunosuppressive activity of uveal melanoma cells, its lymphocytic microenvironment exerts its antitumor effect.Aim of the study. Compare the lymphocytic microenvironment of primary uveal melanomas and distant metastases (to the liver).Мaterials and methods. The tissue material of choroid melanoma after enucleation and the material of tumor metastases for the period 2013-2018 were studied. An immunohistochemical study was performed using CD8, CD4, and CD56 markers for the qualitative and quantitative assessment of lymphocytes in the tumor stroma.Results. Differences were found in the lymphocytic infiltration of the uveal melanoma stroma and its distant metastases. A statistically significantly greater representation of CD4, CD8-lymphocytes and CD56 cells in tumor metastases than in primary melanoma tissue samples, with CD4-lymphocytes predominant. A direct high-strength correlation was registered between the number of CD4-lymphocytes and CD8-lymphocytes.Discussion. Malignant cells actively modify their cellular and stromal-vascular environment, ensuring their active growth and reproduction. The question of the immune reactivity of the surrounding cells in relation to uveal melanoma remains debatable. According to our data, which is consistent with a number of other studies, uveal melanoma cells do not completely evade the body's immune response. Thus, the determination of possible points of antitumor exposure can be based on a detailed study of the microenvironment of uveal melanoma. Conclusions. The pronounced lymphocytic infiltrate found in uveal melanoma metastases in comparison with the primary tumor indicates an active immune response of the body to the tumor. These results of our study confirm the importance of further studying the immune-mediated antitumor effect on uveal melanoma and the need to investigate possible approaches to immunotherapy.

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PROTEIN 24 HIV DAN LIMFOSIT T-CD4+ DI INFEKSI HIV TAHAP I

INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY ◽

10.24293/ijcpml.v21i3.740 ◽

2016 ◽

Vol 21 (3) ◽

pp. 273

Author(s):

I Made Sila Darmana ◽

Endang Retnowati ◽

Erwin Astha Triyono

Keyword(s):

T Lymphocytes ◽

Hiv Infection ◽

T Lymphocyte ◽

Negative Correlation ◽

Stage I ◽

Cross Sectional ◽

Protein Levels ◽

P24 Protein ◽

Persistent Generalized Lymphadenopathy ◽

Spearman Test

Measuring HIV p24 protein is a test which is more practical than determination of CD4+ T-lymphocyte counts and viral load, as it does not require a very sophisticated instrument and requires a lower cost. Independent predictive value of p24 to the decline of CD4+ T-lymphocytes, clinical progression and survival in HIV-infected patients have been reported. In this study, HIV-infected patients were found to have HIV p24 protein levels inversely proportional to CD4+ T-lymphocyte counts by using Spearman test (R2=0.225; p=0.0331). Studies on the correlation between HIV p24 protein levels and CD4+ T-lymphocyte counts in stage I HIV infection have not yet been reported. The aim of this study was to prove the correlation between HIV p24 protein levels and CD4+ T-lymphocytes in stage I HIV infection. Research issue was whether a correlation between HIV p24 protein levels and CD4+ T-lymphocyte counts in stage I HIVinfection existed ? The hypothesis was that a correlation between HIV p24 protein levels and CD4+ T-lymphocyte counts in stage I HIV infection existed. The study design was cross sectional observational. Subjects consisted of 30 stage I HIV-infected patients treated at the Infectious Disease Intermediate Care Unit, Dr. Soetomo Hospital and VCT Clinic of the Dr. Ramelan Naval Hospital, Surabaya from May to July 2014. Stage I HIV infection is an asymptomatic HIV infection or with persistent generalized lymphadenopathy and the patient is able to perform normal activities. Levels of p24 were measured by ELISA method and CD4+ T-lymphocyte counts using flowcytometry(BD FACSCaliburTM). The results were statistically analyzed using Pearson’s correlation test. HIV p24 protein levels in stage I of HIV infection ranged from 1.8 to 10.8 pg/mL, mean of 5.14 pg/mL and a standard deviation of 2.08 pg/mL. CD4+ T-lymphocyte counts decreased with a range of 49-559 cells /uL for absolute values and 4.42–26.02% for percentage values Correlations between blood p24 levels and CD4+ T-lymphocyte counts either absolute (r=–0.392, p=0.032) or percentage (r=–0.363, p=0.049) were found. In stage I HIV-infected patients, a negative correlation was found between p24 levels and CD4+ T-lymphocyte counts, in both CD4+T-lymphocyte counts as absolute and as well as percentage values. This negative correlation showed that the p24 HIV levels were inversely proportional to the CD4+ T-lymphocyte counts. HIV p24 protein levels have a possibility to be used predicting CD4+ T-lymphocyte counts

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Negative Correlation Between Serum Levels of Homocysteine and Apolipoprotein M

Current Molecular Medicine ◽

10.2174/1566524019666190308115624 ◽

2019 ◽

Vol 19 (2) ◽

pp. 120-126

Author(s):

J. Wei ◽

Y. Yu ◽

Y. Feng ◽

J. Zhang ◽

Q. Jiang ◽

...

Keyword(s):

Negative Correlation ◽

Hepg2 Cells ◽

Serum Levels ◽

Significant Negative Correlation ◽

Mrna Levels ◽

Rt Pcr ◽

Apolipoprotein M ◽

Protein Mass ◽

Protein Levels ◽

Phosphoinositide 3 Kinase

Background: Homocysteine (Hcy) has been suggested as an independent risk factor for atherosclerosis. Apolipoprotein M (apoM) is a constituent of the HDL particles. The goal of this study was to examine the serum levels of homocysteine and apoM and to determine whether homocysteine influences apoM synthesis. Methods: Serum levels of apoM and Hcy in 17 hyperhomocysteinemia (HHcy) patients and 19 controls were measured and their correlations were analyzed. Different concentrations of homocysteine (Hcy) and LY294002, a specific phosphoinositide 3- kinase (PI3K) inhibitor, were used to treat HepG2 cells. The mRNA levels were determined by RT-PCR and the apoM protein mass was measured by western blot. Results: We found that decreased serum apoM levels corresponded with serum HDL levels in HHcy patients, while the serum apoM levels showed a statistically significant negative correlation with the serum Hcy levels. Moreover, apoM mRNA and protein levels were significantly decreased after the administration of Hcy in HepG2 cells, and this effect could be abolished by addition of LY294002. Conclusions: resent study demonstrates that Hcy downregulates the expression of apoM by mechanisms involving the PI3K signal pathway.

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TRIM32 and Malin in Neurological and Neuromuscular Rare Diseases

Cells ◽

10.3390/cells10040820 ◽

2021 ◽

Vol 10 (4) ◽

pp. 820

Author(s):

Lorena Kumarasinghe ◽

Lu Xiong ◽

Maria Adelaida Garcia-Gimeno ◽

Elisa Lazzari ◽

Pascual Sanz ◽

...

Keyword(s):

Common Ancestor ◽

Genetic Diseases ◽

Ubiquitin Ligases ◽

E3 Ubiquitin Ligases ◽

Lafora Disease ◽

C Terminus ◽

Rare Genetic Diseases ◽

Tripartite Motif ◽

Trim Proteins ◽

Ring E3

Tripartite motif (TRIM) proteins are RING E3 ubiquitin ligases defined by a shared domain structure. Several of them are implicated in rare genetic diseases, and mutations in TRIM32 and TRIM-like malin are associated with Limb-Girdle Muscular Dystrophy R8 and Lafora disease, respectively. These two proteins are evolutionary related, share a common ancestor, and both display NHL repeats at their C-terminus. Here, we revmniew the function of these two related E3 ubiquitin ligases discussing their intrinsic and possible common pathophysiological pathways.

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Downregulation of survivin expression enhances sensitivity of cultured uveal melanoma cells to cisplatin treatment

Experimental Eye Research ◽

10.1016/j.exer.2005.11.024 ◽

2006 ◽

Vol 83 (1) ◽

pp. 176-182 ◽

Cited By ~ 17

Author(s):

Haochuan Li ◽

Jerry Y. Niederkorn ◽

Sudha Neelam ◽

Hassan Alizadeh

Keyword(s):

Uveal Melanoma ◽

Survivin Expression ◽

Melanoma Cells

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Deregulated FASN Expression in BRAF Inhibitor-Resistant Melanoma Cells Unveils New Targets for Drug Combinations

Cancers ◽

10.3390/cancers13092284 ◽

2021 ◽

Vol 13 (9) ◽

pp. 2284

Author(s):

Serena Stamatakos ◽

Giovanni Luca Beretta ◽

Elisabetta Vergani ◽

Matteo Dugo ◽

Cristina Corno ◽

...

Keyword(s):

Drug Resistance ◽

Metastatic Melanoma ◽

Mutant Cell ◽

Braf Inhibitor ◽

Cancer Cell Line ◽

Melanoma Cells ◽

Melanoma Progression ◽

Increased Sensitivity ◽

Resistant Cells

Metabolic changes promoting cell survival are involved in metastatic melanoma progression and in the development of drug resistance. In BRAF-inhibitor resistant melanoma cells, we explored the role of FASN, an enzyme involved in lipogenesis overexpressed in metastatic melanoma. Resistant melanoma cells displaying enhanced migratory and pro-invasive abilities increased sensitivity to the BRAF inhibitor PLX4032 upon the molecular targeting of FASN and upon treatment with the FASN inhibitor orlistat. This behavior was associated with a marked apoptosis and caspase 3/7 activation observed for the drug combination. The expression of FASN was found to be inversely associated with drug resistance in BRAF-mutant cell lines, both in a set of six resistant/sensitive matched lines and in the Cancer Cell Line Encyclopedia. A favorable drug interaction in resistant cells was also observed with U18666 A inhibiting DHCR24, which increased upon FASN targeting. The simultaneous combination of the two inhibitors showed a synergistic interaction with PLX4032 in resistant cells. In conclusion, FASN plays a role in BRAF-mutated melanoma progression, thereby creating novel therapeutic opportunities for the treatment of melanoma.

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