定量蛋白質(zhì)組學(xué)分析顯示G蛋白偶聯(lián)受體激酶4誘導(dǎo)HepG2細(xì)胞生長抑制

2024年4月，海南省腫瘤醫(yī)院，海南醫(yī)科大學(xué)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(xué)科學(xué)研究中心，中南大學(xué)湘雅二醫(yī)院外科（a Hainan Cancer Hospital, Affiliated Cancer Hospital of Hainan Medical University, Department of Radiotherapy Oncology,

b Hainan Clinical Research Center for Hepatopathy and Liver Critical Illness,

c Guilin Medical University, Center for Science Research,

d Central South University, The Second Xiangya Hospital, Department of Surgery,）

Yan Wang研究團(tuán)隊(duì)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(zhì)組學(xué)分析顯示G蛋白偶聯(lián)受體激酶4誘導(dǎo)HepG2細(xì)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(xì)胞的生物學(xué)作用及其可能的生物學(xué)機(jī)制。

材料與方法

用過表達(dá)grk4的慢病毒載體(OE)和陰性對照慢病毒載體(NC)感染HepG2細(xì)胞，用細(xì)胞計(jì)數(shù)試劑盒-8和流式細(xì)胞術(shù)(FCM)評估細(xì)胞增殖、周期和凋亡。利用定量蛋白質(zhì)組學(xué)技術(shù)分析比較OE細(xì)胞和NC細(xì)胞的蛋白譜和差異表達(dá)蛋白(DEPs)，并利用生物信息學(xué)分析和平行反應(yīng)監(jiān)測(PRM)技術(shù)研究其功能、表達(dá)和可能的機(jī)制。

結(jié)果

接受OE的HepG2細(xì)胞比接受NC的HepG2細(xì)胞生長更慢。流式細(xì)胞儀顯示，與NC細(xì)胞相比，OE細(xì)胞發(fā)生了s期周期阻滯，OE細(xì)胞和NC細(xì)胞均未發(fā)生凋亡。在定量蛋白質(zhì)組學(xué)鑒定的7006個(gè)蛋白中，根據(jù)過濾參數(shù)檢測了403個(gè)DEPs，其中135個(gè)表達(dá)下調(diào)，268個(gè)表達(dá)上調(diào)。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(xì)胞增殖、周期和代謝的生物學(xué)過程。PRM驗(yàn)證了與PPAR通路相關(guān)的DEPs的表達(dá)。

結(jié)論

本研究表明，GRK4可阻止HepG2細(xì)胞增殖，使細(xì)胞周期阻滯在s期，而PPAR通路通過GRK4參與HepG2細(xì)胞的調(diào)控。

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