ZIC1基因过表达激活P53信号通路抑制胸膜间皮瘤细胞增殖

邓勇军; 陈倩; 邹建彬; 宫政; 刘焕鹏

doi:10.12259/j.issn.2095-610X.S20240405

ZIC1基因过表达激活P53信号通路抑制胸膜间皮瘤细胞增殖

doi: 10.12259/j.issn.2095-610X.S20240405

1.
云南大学附属医院胸外科，云南昆明　650021
2.
大理大学公共卫生学院，云南大理　671000

基金项目: 云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目（202001AY0001-089）；云南省兴滇英才支持计划2020年名医专项基金资助项目（YNWR-MY-2020-048）

详细信息

作者简介:
邓勇军（1972～），男，湖北天门人，医学博士，主任医师，主要从事胸外科疾病的微创手术治疗、胸部肿瘤基础与临床研究、胸膜间皮瘤表观遗传学研究工作

通讯作者:
邓勇军， E-mail：thoraces@163.com

中图分类号: R734.3
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出版历程
- 收稿日期: 2023-12-21
- 网络出版日期: 2024-04-07
- 刊出日期: 2024-04-29

Overexpression of ZIC1 Gene Inhibits Proliferation of Pleural Mesothelioma Cells by Activating P53 Signaling Pathway

1.
Dept. of Thoracic Surgery，Affiliated Hospital of Yunnan University ，Kunming Yunnan 650021
2.
School of Public Health，Dali University，Dali Yunnan 671000，China

摘要

摘要: 目的探讨小脑锌指结构1（ZIC1）基因过表达对胸膜间皮瘤SMC-1细胞增殖、凋亡的影响以及相应的机制。方法用携带ZIC1 基因的慢病毒颗粒转染SMC-1细胞作为过表达组，用空载慢病毒颗粒感染SMC-1细胞作为空载体组，将常规培养未进行转染的SMC-1细胞作为空白对照组，采用Western blot在蛋白水平检测3组细胞中ZIC1蛋白的表达情况；采用CCK-8细胞增殖试验检测各组细胞增殖能力，Hoechst-PI双染法检测各组细胞凋亡情况；采用Western blot检测P53介导的细胞凋亡信号通路的主要基因P53、P21、MDM2及P53磷酸化位点Ser392蛋白表达水平。结果与空载组和对照组相比，ZIC1过表达组中ZIC1蛋白的表达明显增高，差异有统计学意义（F = 4.665，P = 0.036）；ZIC1过表达组中肿瘤细胞的增殖活性明显减弱，而肿瘤细胞的凋亡明显增加，差异有统计学意义（P < 0.05）；ZIC1基因过表达组中P53信号通路中的主要基因P53、P21、MDM2及P53-Ser392蛋白表达均明显增加（P < 0.05）。结论 ZIC1基因过表达可以抑制胸膜间皮瘤SMC-1细胞增殖，促进其凋亡，其可能的机制为通过上调P53基因磷酸化位点的表达激活P53基因介导的细胞凋亡信号通路来实现。
- ZIC1 /
- 胸膜间皮瘤细胞 /
- P53通路 /
- 细胞增殖 /
- 细胞凋亡
Abstract: Objective To investigate the effect of cerebellar zinc finger structure 1 （ZIC1） gene overexpression on the proliferation and apoptosis of pleural mesothelioma cells （SMC-1 cells） and the corresponding molecular mechanism. Methods SMC-1 cells were transfected with lentivirus particles carrying ZIC1 gene as the experimental group, infected with empty lentivirus particles as the control group, and conventional cultured untransfected SMC-1 cells as the blank control group. Western blot was used to detect the expression of ZIC1 protein in the three groups. CCK-8 cell proliferation assay was used to detect the proliferation ability of each group, and Hoechst-PI double staining was used to detect the apoptosis of each group. The protein expression levels of P53, P21, MDM2 and P53 phosphorylation site Ser392 in P53-mediated apoptosis signaling pathway were detected by Western blot. Results Compared with the empty vector group and the control group, the expression of ZIC1 protein in the ZIC1 overexpression group was significantly increased, with a statistically significant difference （F = 4.665, P = 0.036）. Compared with the control group, the proliferation ability of SMC-1 cells in the ZIC1 overexpression group was significantly reduced, while the apoptosis of tumor cells was significantly increased, with a statistically significant difference （P < 0.05）. The main genes P53, P21, MDM2, and P53 Ser392 protein expression in the P53 signaling pathway in the ZIC1 gene overexpression group were significantly increased. Conclusion Overexpression of ZIC1 gene can inhibit the proliferation of pleural mesothelioma SMC-1 cells and promote their apoptosis. The possible mechanism is to activate the P53 gene mediated apoptosis signaling pathway by upregulating the expression of P53 gene phosphorylation sites.
- ZIC1 /
- Pleural mesothelioma cells /
- P53 signal pathway /
- Proliferation /
- Apoptosis

HTML全文

图 1 ZIC1在各组细胞中的蛋白表达情况

Mean±SD，n = 3，^*P < 0.05。

Figure 1. The ZIC1 protein expression in tumor cells of each group

下载: 全尺寸图片幻灯片

图 2 3组肿瘤细胞的增殖能力比较

Mean±SD，n = 3，^*P < 0.05。

Figure 2. The ability of cell proliferation of three groups

下载: 全尺寸图片幻灯片

图 3 3组胸膜间皮瘤细胞凋亡情况（×200）

采用HO-PI染色进行检测，红色荧光细胞为晚期凋亡或死亡细胞，蓝色荧光较亮的细胞为早期凋亡细胞，其他蓝色荧光强度较弱的细胞显示为活细胞和自然细胞。

Figure 3. Apoptosis of pleural mesothelioma cells in the 3 groups （×200）

下载: 全尺寸图片幻灯片

图 4 MDM2、P21、P53及P53（Ser392）在各组细胞中的蛋白表达情况

Mean±SD，n = 3，^*P < 0.05;^**P < 0.01;^*** P < 0.001。

Figure 4. The proteins expression of MDM2、P21、P53 and P53（Ser392） in each group of cells

下载: 全尺寸图片幻灯片

表 1 ZIC1在各组细胞中的蛋白表达

Table 1. The ZIC1 protein expression in tumor cells of each group

组别	ZIC1过表达组	空载体组	空白组	F	P
相对表达量	0.785±0.021	0.373±0.019	0.358±0.022	4.665	0.036^*
^*P < 0.05。

下载: 导出CSV

表 2 各实验组在不同时间点的 OD值

Table 2. The optical density value of each group cell at different time points

分组	8 h	12 h	24 h	36 h	48 h	72 h
ZIC1过表达组	0.677±0.016	0.788±0.018	0.913±0.021	0.988±0.017	1.023±0.022	1.165±0.020
空载体组	0.798±0.022	0.878±0.024	1.233±0.023	1.355±0.016	2.238±0.023	3.266±0.018
空白组	0.836±0.023	0.955±0.027	1.311±0.026	1.402±0.022	2.548±0.028	3.476±0.023

下载: 导出CSV

参考文献(19)

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