[The Effects of IGHG1 on the Proliferation and Apoptosis of Human Acute Myeloid Leukemia THP-1 Cells].

转染 THP1细胞系 分子生物学 髓系白血病 细胞凋亡 细胞周期 流式细胞术 细胞生长 信号转导 生物 SMAD公司 细胞培养 化学 癌症研究 细胞生物学 生物化学 遗传学
作者
Xin Gao,Li-Jing Chu,Tianping Chen
出处
期刊:PubMed 卷期号:31 (5): 1263-1271
标识
DOI:10.19746/j.cnki.issn.1009-2137.2023.05.002
摘要

To investigate the effects of the immunoglobulin G1 heavy chain constant region (IGHG1) on the proliferation and apoptosis of acute myeloid leukemia (AML) THP-1 cells and its possible mechanism of action.Human AML THP-1 cells were cultured in vitro and divided into control (normally cultured THP-1 cells), pcDNA3.1 [THP-1 cells transfected with IGHG1 overexpression (pcDNA3.1-IGHG1) negative control plasmid], pcDNA3.1-IGHG1 (THP-1 cells transfected with plasmid containing pcDNA3.1-IGHG1), LY364947 [transforming growth factor-β (TGF-β)/signal transduction protein (Smad) inhibitor LY364947 20 μmol/L treated THP-1 cells], si-NC [THP-1 cells transfected with IGHG1-small interfering RNA (siRNA) negative control], si-IGHG1 (THP-1 cells transfected with IGHG1-siRNA), and si-IGHG1+LY364947 (IGHG1-siRNA and LY364947 co-treated THP-1 cells) a total of 7 groups. Fluorescence quantitative PCR was used to detect the expression of IGHG1 and immunoglobulin G (IgG) mRNA of THP-1 cells in each group; CCK-8 was used to detect THP-1 cells proliferation activity; flow cytometry was used to detect THP-1 cells apoptosis and cell cycle in each group; Western blot was used to detect the THP-1 cells proliferation, apoptosis and the expression of TGF-β/Smad signaling pathway related proteins.Compared with the control group, after overexpression of IGHG1, the expression of IGHG1 and IgG mRNA, cell proliferation viability, S phase cell ratio, expressions of Cyclin D1, B cell lymphoma-2 (Bcl-2), IgG, TGF-β1, phosphorylated Smad3 (p-Smad3)/Smad3 protein in THP-1 cells were significantly increased (P<0.05), the apoptosis rate, G0/G1 phase cell ratio, expression of p21, Bcl-2 related X protein (Bax), Caspase-3 protein were significantly reduced (P<0.05); after inhibiting TGF-β/Smad signaling pathway or silencing IGHG1, the expression of IGHG1 and IgG mRNA, cell proliferation viability, S phase cell ratio, expression of Cyclin D1, Bcl-2, IgG, TGF-β1, p-Smad3/Smad3 protein in THP-1 cells were significantly reduced (P<0.05), the apoptosis rate, G0/G1 phase cell ratio, expressions of p21, Bax, and Caspase-3 protein were significantly increased (P<0.05); and compared with silencing IGHG1, after co-treatment of IGHG1 gene silencing and TGF-β/Smad pathway inhibition, the expression of IGHG1 and IgG mRNA, cell proliferation viability, S phase cell ratio, expressions of Cyclin D1, Bcl-2, IgG, TGF-β1, p-Smad3/Smad3 protein in THP-1 cells were significantly reduced (P<0.05), the apoptosis rate, G0/G1 phase cell ratio, expression of p21, Bax, and Caspase-3 protein were significantly increased (P<0.05).Silencing IGHG1 gene can down-regulate the expression of IgG, inhibit the proliferation of human AML THP-1 cells, block cell cycle progression, and induce cell apoptosis; its mechanism may be related to the inhibition of the TGF-β/Smad pathway activation.IGHG1对人急性髓系白血病THP-1细胞增殖、凋亡的影响.探讨免疫球蛋白G1重链恒定区(IGHG1)对急性髓系白血病(AML)细胞系THP-1细胞增殖、凋亡的影响及其可能的作用机制.体外培养人AML THP-1细胞,分为对照(正常培养的THP-1细胞)、pcDNA3.1[转染IGHG1过表达(pcDNA3.1-IGHG1)阴性对照质粒的THP-1细胞]、pcDNA3.1-IGHG1(转染pcDNA3.1-IGHG1的THP-1细胞)、LY364947[转化生长因子-β(TGF-β)/信号转导蛋白(Smad)抑制剂LY364947 20 μmol/L处理THP-1细胞)]、si-NC[转染IGHG1小干扰RNA(IGHG1-siRNA)阴性对照的THP-1细胞]、si-IGHG1(转染IGHG1-siRNA的THP-1细胞)和si-IGHG1+LY364947(IGHG1-siRNA和LY364947共同处理THP-1细胞)共7组。荧光定量PCR法检测各组THP-1细胞中IGHG1和免疫球蛋白G(IgG) mRNA的表达;CCK-8法检测各组THP-1细胞增殖活力;流式细胞术检测各组THP-1细胞凋亡率和细胞周期变化;蛋白印迹法检测各组THP-1细胞增殖、凋亡及TGF-β/Smad信号通路相关蛋白的表达.与对照组相比,过表达IGHG1后THP-1细胞中IGHG1和IgG mRNA表达、细胞增殖活力、S期的细胞比例、细胞周期蛋白D1(Cyclin D1)、B细胞淋巴瘤-2(Bcl-2)、IgG、TGF-β1、磷酸化Smad3(p-Smad3)/Smad3蛋白表达均显著升高(P<0.05),细胞凋亡率、G0/G1期的细胞比例、p21、Bcl-2相关X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)蛋白表达均显著降低(P<0.05)。抑制TGF-β/Smad信号通路或沉默IGHG1后THP-1细胞中IGHG1和IgG mRNA表达、细胞增殖活力、S期的细胞比例、Cyclin D1、Bcl-2、IgG、TGF-β1、p-Smad3/Smad3蛋白表达均显著降低(P<0.05),细胞凋亡率、G0/G1期的细胞比例、p21、Bax、Caspase-3蛋白表达均显著升高(P<0.05);且与沉默IGHG1相比,IGHG1基因沉默和TGF-β/Smad通路抑制共同处理的THP-1细胞中IGHG1和IgG mRNA表达、细胞增殖活力、S期的细胞比例、Cyclin D1、Bcl-2、IgG、TGF-β1、p-Smad3/Smad3蛋白表达均显著降低(P<0.05),细胞凋亡率、G0/G1期的细胞比例、p21、Bax、Caspase-3蛋白表达均显著升高(P<0.05).沉默IGHG1基因可下调IgG的表达,抑制人AML THP-1细胞增殖,阻滞细胞周期进程,并诱导细胞凋亡;其作用机制可能与抑制TGF-β/Smad通路的激活有关.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
英姑应助聪慧的正豪采纳,获得10
1秒前
2秒前
整齐荟完成签到,获得积分10
2秒前
领导范儿应助威武的元冬采纳,获得30
6秒前
6秒前
junjun2011完成签到,获得积分10
8秒前
Dasha完成签到,获得积分10
8秒前
充电宝应助puzi采纳,获得10
10秒前
LiuLiu发布了新的文献求助10
11秒前
深情安青应助昏睡的绿海采纳,获得10
11秒前
12秒前
大大的西瓜完成签到 ,获得积分10
16秒前
华仔应助XYN1采纳,获得10
17秒前
17秒前
A阿澍发布了新的文献求助30
17秒前
丘比特应助TMX采纳,获得10
19秒前
21秒前
量子星尘发布了新的文献求助10
22秒前
24秒前
无奈满天发布了新的文献求助10
27秒前
27秒前
27秒前
28秒前
29秒前
30秒前
zhou269完成签到,获得积分10
32秒前
在逃公主许翠花完成签到,获得积分10
32秒前
littleblack发布了新的文献求助10
33秒前
TMX发布了新的文献求助10
33秒前
33秒前
DaiTing发布了新的文献求助10
33秒前
34秒前
简单点发布了新的文献求助10
34秒前
华仔应助无奈满天采纳,获得10
34秒前
fjejj发布了新的文献求助10
35秒前
sisi发布了新的文献求助10
36秒前
华仔完成签到,获得积分10
38秒前
华仔应助特梅头采纳,获得10
40秒前
41秒前
无奈满天完成签到,获得积分10
42秒前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
‘Unruly’ Children: Historical Fieldnotes and Learning Morality in a Taiwan Village (New Departures in Anthropology) 400
Indomethacinのヒトにおける経皮吸収 400
Phylogenetic study of the order Polydesmida (Myriapoda: Diplopoda) 370
基于可调谐半导体激光吸收光谱技术泄漏气体检测系统的研究 350
Robot-supported joining of reinforcement textiles with one-sided sewing heads 320
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3988920
求助须知:如何正确求助?哪些是违规求助? 3531290
关于积分的说明 11253247
捐赠科研通 3269903
什么是DOI,文献DOI怎么找? 1804830
邀请新用户注册赠送积分活动 882027
科研通“疑难数据库(出版商)”最低求助积分说明 809052