PM2.5对L02肝细胞部分癌基因和凋亡相关基因表达的影响

doi:10.3969/j.issn.1004-616x.2020.04.006

癌变·畸变·突变 ›› 2020, Vol. 32 ›› Issue (4): 281-285.doi: 10.3969/j.issn.1004-616x.2020.04.006

PM_2.5对L02肝细胞部分癌基因和凋亡相关基因表达的影响

秦双建^1,2, 李柏茹^1,2, 蔡颖^2,3, 郑凯^2,3, 王冰玉^2,3, 李闰冰^2,3, 肖芳¹, 曾明¹, 徐新云²

1. 中南大学湘雅公共卫生学院, 湖南长沙 410078;
2. 深圳市疾病预防控制中心, 广东深圳 518055;
3. 南华大学公共卫生学院, 湖南衡阳 421001

收稿日期:2020-05-08 修回日期:2020-06-28 出版日期:2020-07-31 发布日期:2020-08-01
通讯作者: 曾明,E-mail:zengming@csu.edu.cn;徐新云,E-mail:xyxu2008@163.com E-mail:zengming@csu.edu.cn;xyxu2008@163.com
作者简介:秦双建,E-mail:1375195449@qq.com。
基金资助:
深圳市科技研发项目（JCYJ20170413101713324，JCYJ20190807102205480）

PM_2.5 exposure on expression of oncogenes and apoptosis genes in hepatocytes

QIN Shuangjian^1,2, LI Boru^1,2, CAI Ying^2,3, ZHENG Kai^2,3, WANG Bingyu^2,3, LI Runbing^2,3, XIAO Fang¹, ZENG Ming¹, XU Xinyun²

1. Xiangya School of Public Health, Central South University, Changsha 410078, Hunan;
2. Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, Guangdong;
3. School of Public Health, University of South China, Hengyang 421001, Hunan, China

Received:2020-05-08 Revised:2020-06-28 Online:2020-07-31 Published:2020-08-01

摘要/Abstract

摘要： 目的： 探讨细颗粒物（PM_2.5）对人正常肝细胞（L02肝细胞）癌基因和凋亡相关基因表达的影响。方法： 以L02肝细胞为研究对象，设10和50 μg/mL两个PM_2.5混悬液染毒组，以未染毒的L02肝细胞为阴性对照，10 μmol/L的K₂CrO₄（下用Cr⁶⁺表示）染毒细胞为阳性对照，均处理24 h，应用实时荧光定量聚合酶链式反应（qPCR）和Western blot技术分别检测促癌基因c-myc、c-fos、抑癌基因p53和促凋亡相关基因Caspase-3、Caspase-8，抑凋亡基因Bcl-2的mRNA以及相应的蛋白表达变化。结果： qPCR法分析结果显示，与阴性对照组比较，10、50 μg/mL PM_2.5混悬液染毒组以及阳性对照组L02肝细胞癌基因c-myc的mRNA表达水平分别升高69.5%、118.0%、51.1%，c-fos表达水平分别升高50.3%、64.4%、23.3%，p53表达水平分别下降4.0%、22.0%、18.0%；凋亡相关基因Caspase-3的mRNA表达水平分别升高31.0%、30.5%、42.0%，Caspase-8表达分别升高25.3%、40.3%、64.5%，Bcl-2表达分别下降40.5%、46.9%、41.4%，差异均有统计学意义（P < 0.05）。Western blot法分析结果显示，在10、50 μg/mL PM_2.5混悬液和Cr⁶⁺染毒后L02肝细胞c-myc蛋白表达水平分别升高32.3%、49.3%、70.6%，c-fos蛋白表达水平分别升高11.3%、50.2%、45.2%，p53蛋白表达水平分别下降17.5%、40.0%、42.9%；Caspase-3蛋白表达水平分别升高23.1%、33.9%、43.1%，Caspase-8蛋白表达水平分别升高31.4%、52.1%、80.0%，Bcl-2蛋白表达水平分别下降14.3%、23.3%、36.9%，差异均有统计学意义（P < 0.05）。结论： PM_2.5染毒引起L02肝细胞中促癌基因和促凋亡相关基因表达升高，抑癌基因和抑凋亡相关基因表达下降，提示PM_2.5对L02肝细胞的恶性转化和凋亡可能有促进与激活作用。

关键词: 大气细颗粒物, L02肝细胞, 癌基因, 凋亡相关基因, 基因表达

Abstract: OBJECTIVE: To study the effect of particulate matter 2.5(PM_2.5) exposure on expression of oncogenes and apoptosis-related genes in normal hepatocytes (L02 cells). METHODS: L02 cells were exposed to PM_2.5 (10 and 50 μg/mL), as the experimental group, to 10 μmol/L Cr⁶⁺ as the positive control group, and nothing as the negative control group,for 24 h. After the treatment,mRNA expression of oncogenes including promoting oncogenes including c-myc, c-fos and suppressing oncogenes p53 and promoting apoptosis-related genes including Caspase-3, Caspase-8 and suppressing apoptosis gene Bcl-2 were detected by fluorescent quantitative real-time PCR(qPCR),the protein expression of oncogenes and apoptosis genes were detected with Western blot. RESULTS: The qPCR showed that compared with the negative control group, significantly different mRNA expressions in the PM_2.5 10 μg/mL,50 μg/mL and positive control groups were:c-myc gene increased by 69.5%,118.0%,51.1%,c-fos gene increased by 50.3%,64.4%,23.3%;p53 gene decreased by 4.0%,22.0%,18.0%;Caspase-3 gene increased by 31.0%,30.5% and 42.0%;Caspase-8 expression level increased by 25.3%,40.3% and 64.5%,Bcl-2 gene expression level decreased by 40.5%,46.9% and 41.4%, respectively (P < 0.05). Western blot analyses showed that expression levels of c-myc protein increased by 32.2%, 49.3%,70.6%;c-fos protein increased by 11.3%,50.2%,45.2%;p53 protein decreased by 17.5%,40.0%, 42.5%;Caspase-3 protein increased by 23.1%,33.9%,43.1%;Caspase-8 protein increased by 31.4%,52.1%, 80.0%; Bcl-2 protein decreased by 14.3%, 23.3%, 36.9%, respectively, (P < 0.05). CONCLUSION: PM_2.5 exposure increased expression of promoting oncogenes and promoting apoptosis genes as well as decreased expression of suppressing oncogenes and suppressing apoptosis genes in L02 hepatocytes. The results indicate that PM_2.5 exposure can promote and activate malignant transformation and apoptosis of L02 hepatocytes.

Key words: PM_2.5, L02 hepatocytes, oncogenes, apoptosis-related genes, gene expression

中图分类号:

R122.7

秦双建, 李柏茹, 蔡颖, 郑凯, 王冰玉, 李闰冰, 肖芳, 曾明, 徐新云. PM_2.5对L02肝细胞部分癌基因和凋亡相关基因表达的影响[J]. 癌变·畸变·突变, 2020, 32(4): 281-285.

QIN Shuangjian, LI Boru, CAI Ying, ZHENG Kai, WANG Bingyu, LI Runbing, XIAO Fang, ZENG Ming, XU Xinyun. PM_2.5 exposure on expression of oncogenes and apoptosis genes in hepatocytes[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(4): 281-285.

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PM_2.5对L02肝细胞部分癌基因和凋亡相关基因表达的影响

PM_2.5 exposure on expression of oncogenes and apoptosis genes in hepatocytes

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PM2.5对L02肝细胞部分癌基因和凋亡相关基因表达的影响

PM2.5 exposure on expression of oncogenes and apoptosis genes in hepatocytes

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PM_2.5对L02肝细胞部分癌基因和凋亡相关基因表达的影响

PM_2.5 exposure on expression of oncogenes and apoptosis genes in hepatocytes