Dataset Information

Proteomics reveals the inhibitory mechanism of Levodopa against esophageal squamous cell carcinoma

ABSTRACT: The high recurrence rate and poor survival prospects of esophageal squamous cell carcinoma (ESCC) patients after treatment make the ongoing research on chemoprevention drugs for ESCC particularly important. In helping to solve this problem, we screened a large number of FDA-approved drugs and found that levodopa, a drug used to treat Parkinson’s disease, has an inhibitory effect on the growth of ESCC cells. To elucidate the molecular mechanisms, we applied mass spectrometry to investigate the anti-tumor activity of levodopa on ESCC. The results suggest that levodopa can down-regulate oxidative phosphorylation, non-alcoholic fatty liver disease (NAFLD) and parkinson disease pathways. SDHD, NDUFS4 and MT-CO3, major respiratory compounds in the mitochondria, were involved in these pathways. Down-regulation of these proteins is associated with mitochondrial dysfunction. Western blotting and immunofluorescence results confirmed the authenticity of proteomics data. Cell viability assay revealed that mitochondrial activity had been suppressed after levodopa treatment. Mitochondrial membrane potential reduction was detected by JC-1 and TMRE assays. And transmission electron microscope (TEM) analysis indicated that mitochondrial morphology changed. Taken together, levodopa inhibits the growth of ESCC through restraining the mitochondria.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: xiaomei li

LAB HEAD: Kangdong Liu

PROVIDER: PXD013824 | Pride | 2021-09-08

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

Items per page:

1 - 5 of 13

Publications

Proteomics Reveal the Inhibitory Mechanism of Levodopa Against Esophageal Squamous Cell Carcinoma.

Li Zhenzhen Z Li Xin X He Xinyu X Jia Xuechao X Zhang Xiaofan X Lu Bingbing B Zhao Jimin J Lu Jing J Chen Lexia L Dong Ziming Z Liu Kangdong K Dong Zigang Z

Frontiers in pharmacology 20200925

High recurrence rates and poor survival of patients with esophageal squamous cell carcinoma (ESCC) after treatment make ongoing research on chemoprevention drugs for ESCC particularly important. In this study, we screened a large number of FDA-approved drugs and found levodopa, a drug used to treat Parkinson's disease, had an inhibitory effect on the growth of ESCC cells. To elucidate the molecular mechanisms involved, we applied quantitative proteomics to investigate the anti-tumor activity of ...[more]

PMID: 33101026

Similar Datasets

Project description:The management of cardiac ischemic injury has been challenged by ischemia and reperfusion (I/R) injury. Reactive oxygen species (ROS) generated from mitochondrial reverse electron transport (RET) during the early phase of reperfusion is considered to be the initiating cause for ischemia and reperfusion injury. Ginsenosides and their prescriptions are widely used in the clinic for treatment of myocardial ischemia, however, the action to combat ROS remains to be elucidated. In this work, we used TMT-based proteomic approach to detect differential proteins from mitochondrial fractions of mice hearts with ischemia-reperfusion. Results indicated that the mass error of identified peptides was within 10 ppm, and most of the identified peptides were composed of 7-23 amino acids. Using these qualified data, 17,262 peptides, with a confidence level ≥ 95%, were mapped to 3,054 protein groups. PCA analysis showed that the model group was clearly separated from the blank group, while the protein pattern was partly reversed with Rb1 treatment. With a criteria of p-value < 0.05, 591 significantly changed proteins including 186 increased and 405 decreased ones in the model group were identified compared with the blank group. These proteins were divided into 4 clusters. Proteins in Cluster I highly increased in the model group, but clearly decreased in the Rb1 group. Proteins in Cluster IV clearly decreased in the model group, but markedly increased in the Rb1 group. Proteins in Cluster III and in Cluster IV were not significantly or slightly regulated by the Rb1 treatment. GO analysis of Cluster I and II indicated that the molecular function of these proteins were closely related to oxidoreductase activity and NADH dehydrogenase activity. Our result showed that Rb1 administration before ischemia markedly decreased infarct size (48 h post-I/R), and preserved cardiac function (2 weeks post-I/R), and subsequently limited tissue fibrosis (28 days post-I/R). These results indicated that targeted inhibition of mitochondrial complex I in the early stage of reperfusion by Rb1 is a potential therapeutic strategy for alleviating IR injury. This work not only indicates a potent molecular target for the precision therapy of myocardial ischemia by Rb1, but also provides novel knowledge for the management of cardiac ischemic injury by traditional Chinese medicine.

Project description:Abstract Rationale: Interferon (IFN) plays a key role in immunotherapy by acting through interferon-stimulated genes. Interferon resistance limits its efficacy. Ubiquitination modification of key proteins is a popular research direction in hepatocellular carcinoma (HCC), and it is also one of the important mechanisms of interferon resistance. This study is dedicated to discovering the link between the pathogenesis of interferon resistance and ubiquitination modifications in HCC. Methods: We first combined the TCGA LIHC dataset and genes from the iUUCD 2.0 database to screen the most promising ubiquitination-related gene, UBE2O, in HCC using a bioinformatics approach. A combination of proteomic analysis, mass spectrometry, and survival analysis was used to screen for pathways and proteins negatively regulated by UBE2O. The effect of silencing UBE2O on interferon sensitivity in HCC cells was assessed using in vitro clone formation, migration, and wound healing assays and in vivo subcutaneous tumor xenograft models. Changes in the half-life of substrates following inhibition of UBE2O were assessed using a cycloheximide method. Two databases were used to predict ubiquitination sites on IFIT3. After rescue experiments to exclude the effect of substrate, proteomic analysis was reperformed to analyze the effect on the interferon pathway and phenotype. The effect on substrate and interferon sensitivity was observed using arsenic trioxide (ATO) to inhibit UBE2O. Results: Initially, UBE2O was deduced from 807 UUCRGs in the TCGA LIHC dataset by six consecutive screening steps. UBE2O was significantly overexpressed in HCC cell lines and tissues compared to the corresponding normal group, and low expression of UBE2O was associated with a better prognosis. UBE2O was knocked down for proteomic analysis, and the results suggested that UBE2O negatively regulates the interferon-alpha/beta signaling (p＜0.01) and the interferon pathway (p＜0.01). IFIT3 in the interferon pathway was identified as a possible ubiquitinated substrate of UBE2O by combining mass spectrometry, the human protein atlas (HPA), and TCGA-based survival analysis. There was a significant negative correlation between UBE2O and IFIT3, including survival analysis showing opposite trends and a negative correlation in protein expression. UBE2O binds to IFIT3, and interferon-α promotes this binding. The stability of IFIT3 protein increased with decreasing UBE2O (p＜0.05), and these effects disappeared with the mutational inactivation of UBE2O. K236 of IFIT3 is the active site of ubiquitination. By increasing IFIT3, the knockdown of UBE2O increases the sensitivity of HCC cells to interferon, leading to reduced proliferation and migration (all p＜0.05). These effects disappeared after the elimination of IFIT3. Additionally, ATO inhibited UBE2O and therefore increased interferon sensitivity in HCC cells. Conclusions: UBE2O exerts a protumor role and targets a new substrate, IFIT3, for ubiquitination and degradation, impeding the interferon pathway activation and interferon sensitization in HCC. The findings will shed light on the interferon-based targeted or combination therapeutic strategies for HCC in the future.

			Action	DRS
	rep1_71203LQ_Ctrl-01.raw	Raw
	rep1_71203LQ_Ctrl-02.raw	Raw
	rep1_71203LQ_Ctrl-03.raw	Raw
	rep1_71203LQ_Ctrl-04.raw	Raw
	rep1_71203LQ_Ctrl-05.raw	Raw