Quantitative proteomics assay reveals G protein-coupled receptor kinase 4-induced HepG2 cell growth inhibition

2024年4月，海南省腫瘤醫(yī)院，海南醫(yī)科大學附屬腫瘤醫(yī)院，放射腫瘤科，海南省肝病與肝臟危重癥臨床研究中心，桂林醫(yī)科大學科學研究中心，中南大學湘雅二醫(yī)院外科（Hainan Cancer Hospital, Affiliated Cancer Hospital of Hainan Medical University, Department of Radiotherapy Oncology,

Hainan Clinical Research Center for Hepatopathy and Liver Critical Illness,

Guilin Medical University, Center for Science Research,

dCentral South University, The Second Xiangya Hospital, Department of Surgery,）

Yan Wang研究團隊在《Heliyon》上發(fā)表論文：

“Quantitative proteomics assay reveals G protein-coupled receptor kinase 4-induced HepG2 cell growth inhibition”

“定量蛋白質組學分析顯示G蛋白偶聯(lián)受體激酶4誘導HepG2細胞生長抑制”

Abstract:

Background and aim: To investigate the biological effects and putative biological mechanism of Gprotein-coupled receptor kinase 4 (GRK4) on HepG2 cells.

Materials and methods: Cell proliferation, cycle, and apoptosis were evaluated by Cell Counting Kit-8 and flow cytometry (FCM) in HepG2 cells infected with either the GRK4-overexpressing lentivirus vector (OE) or the negative control lentivirus vector (NC). The protein profiles and differentially expressed proteins (DEPs) of the OE and NC cells were analyzed and compared using the quantitative proteomics technique, and their function, expression, and probable mechanism were investigated using bioinformatic assays and parallel reaction monitoring (PRM).

Results: HepG2 cells that received the OE grew more slowly than those that received the NC. FCM revealed that, when compared to the NC cells, the OE cells had undergone S-phase cycle arrest, and neither the OE nor NC cells underwent apoptosis. Among the 7006 proteins that were identified by quantitative proteomics, 403 DEPs were examined based on the filtering parameters, with the expressions of 135 being downregulated and 268 being upregulated. In addition to being involved in the peroxisome proliferator-activated receptor (PPAR) signaling pathway, the DEPs

were implicated in the biological processes of cell proliferation, cycle, and metabolism. PRM verified the expressions of DEPs that were connected to the PPAR pathway.

Conclusions: This study shows that GRK4 prevents HepG2 cells from proliferating and causes cell

cycle arrest in the S-phase, while the PPAR pathway is involved in the regulation of HepG2 cells via GRK4.

摘要：

背景與目的

探討G蛋白偶聯(lián)受體激酶4 (GRK4)對HepG2細胞的生物學作用及其可能的生物學機制。

材料與方法

用過表達grk4的慢病毒載體(OE)和陰性對照慢病毒載體(NC)感染HepG2細胞，用細胞計數(shù)試劑盒-8和流式細胞術(FCM)評估細胞增殖、周期和凋亡。利用定量蛋白質組學技術分析比較OE細胞和NC細胞的蛋白譜和差異表達蛋白(DEPs)，并利用生物信息學分析和平行反應監(jiān)測(PRM)技術研究其功能、表達和可能的機制。

結果

接受OE的HepG2細胞比接受NC的HepG2細胞生長更慢。流式細胞儀顯示，與NC細胞相比，OE細胞發(fā)生了s期周期阻滯，OE細胞和NC細胞均未發(fā)生凋亡。在定量蛋白質組學鑒定的7006個蛋白中，根據過濾參數(shù)檢測了403個DEPs，其中135個表達下調，268個表達上調。除了參與過氧化物酶體增殖物激活受體(PPAR)信號通路外，DEPs還參與細胞增殖、周期和代謝的生物學過程。PRM驗證了與PPAR通路相關的DEPs的表達。

結論

本研究表明，GRK4可阻止HepG2細胞增殖，使細胞周期阻滯在s期，而PPAR通路通過GRK4參與HepG2細胞的調控。

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