Upregulation of Id3 inhibits cell proliferation and induces apoptosis in A549/DDP human lung cancer cells in vitro
Upregulation of Id3 inhibits cell proliferation and induces apoptosis in A549/DDP human lung cancer cells in vitro
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DOI：
10.3892/mmr.2016.5221
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发表时间：
2016-07-01
影响因子：
3.4
通讯作者：
Li, Xiaojun
中科院分区：
文献类型：
--
作者：
Chen, Fangfang;Zhao, Qinfei;Li, Xiaojun
DNA结合抑制因子（Inhibitor of DNA binding，Id）3是Id多基因家族中的一员，是一种显性负螺旋-环-螺旋转录因子，在人类肿瘤中起癌基因或肿瘤抑制因子的作用。它的上调最近被证明对肺癌具有抑制作用，肺癌是全球癌症相关死亡率的主要原因。由于耐药性是癌症治疗的主要瓶颈，本研究评估了Id 3抑制顺铂耐药的A549肺腺癌细胞（A549/DDP）的能力。用增强型绿色荧光蛋白过表达质粒（pEGFP）和Id 3过表达质粒（Id 3/pEGFP）瞬时转染A549/DPP细胞，经荧光共聚焦显微镜、PCR和western blot检测证实转染成功。采用MTT法、Hoechst 33258荧光显微镜和Annexin V/propidium iodide双染流式细胞仪检测Id 3对A549/DDP细胞活力和凋亡的影响。结果显示，Id 3过表达可明显抑制A549/DDP细胞的增殖和活力，并呈时间依赖性。此外，Id 3的过表达使A549/DDP细胞的凋亡率从2.73%显著增加到16.92%，证实了Id 3在肿瘤生长的阴性对照中的意义。本研究的结果表明，Id 3的过表达可能作为一种新的策略，抑制顺铂敏感的肺癌。Id 3过表达是否能增强肺癌细胞对顺铂的敏感性，将进行进一步的实验研究。
Inhibitor of DNA binding (Id) 3 is a member of the Id multigene family of dominant-negative helix-loop-helix transcription factors, which function as oncogenes or tumor suppressors in human cancers. Its upregulation was recently shown to have inhibitory effects on lung cancer, which is the leading cause of cancer-associated mortality worldwide. As drug resistance represents a major bottleneck of cancer therapy, the present study assessed the ability of Id3 to inhibit cisplatin-resistant A549 lung adenocarcinoma cells (A549/DDP). A549/DPP cells were transiently transfected with enhanced green fluorescence protein overexpression plasmid (pEGFP) or Id3 overexpression plasmid (Id3/pEGFP), which was confirmed by confocal fluorescence microscopy, PCR and western blot analysis. The effects of Id3 on the viability and apoptosis of A549/DDP were determined using an MTT assay, fluorescence microscopy with Hoechst 33258 staining and flow cytometry following Annexin V/propidium iodide double staining. The results revealed that overexpression of Id3 significantly inhibited the proliferation and viability of A549/DDP cells in a time-dependent manner. Furthermore, overexpression of Id3 significantly increased the apoptotic rate of A549/DDP cells from 2.73 to 16.92%, confirming the implication of Id3 in the negative control of tumour growth. The results of the present study revealed that overexpression of Id3 may serve as a novel strategy for inhibiting cisplatin-sensitive lung cancer. Further experiments will be performed to determine whether Id3 overexpression could enhance the sensitivity of lung cancer cells to DDP.