甲基丙烯酸缩水甘油酯通过ERK/MMP14信号通路影响16HBE细胞恶性转化的研究

doi:10.3969/j.issn.1004-616x.2024.04.003

癌变·畸变·突变 ›› 2024, Vol. 36 ›› Issue (4): 261-267.doi: 10.3969/j.issn.1004-616x.2024.04.003

• 论著 • 上一篇

甲基丙烯酸缩水甘油酯通过ERK/MMP14信号通路影响16HBE细胞恶性转化的研究

崔旭芳^1,2, 王全凯^1,2, 金惠萍^1,2, 李昕苇^1,2, 顾轶婷¹, 乌瀚宝栎尔¹, 康同影¹, 许建宁^1,2

1. 中国疾病预防控制中心职业卫生与中毒控制所, 北京 100050;
2. 中国疾病预防控制中心创伤与化学中毒全国重点实验室, 北京 100050

收稿日期:2023-12-29 修回日期:2024-06-12 发布日期:2024-08-06
通讯作者: 许建宁
作者简介:崔旭芳，E-mail：cuixufang2022@163.com。
基金资助:
国家自然科学基金(81673221)

Glycidyl methacrylate affects malignant transformation of 16HBE cells through the ERK/MMP14 signaling pathway

CUI Xufang^1,2, WANG Quankai^1,2, JIN Huiping^1,2, LI Xinwei^1,2, GU Yiting¹, WUHAN Baolier¹, KANG Tongying¹, XU Jianning^1,2

1. National Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention, Beijing 100050;
2. State Key Laboratory of Trauma and Chemical Poisoning, Chinese Center for Disease Control and Prevention, Beijing 100050, China

Received:2023-12-29 Revised:2024-06-12 Published:2024-08-06

摘要/Abstract

摘要： 目的：探讨甲基丙烯酸缩水甘油酯(GMA)是否通过ERK/MMP14信号通路影响人支气管上皮(16HBE)细胞的恶性转化，为进一步探究GMA诱导16HBE细胞恶性转化的可能分子机制提供线索。方法：8 μg/mL的GMA重复染毒16HBE细胞为GMA处理组，等体积二甲基亚砜(DMSO)处理细胞作为溶剂对照组，处理后的细胞传代培养。收获第40代恶性转化16HBE细胞，采用软琼脂集落形成实验确证细胞的恶性转化程度；细胞划痕实验和Transwell细胞迁移实验检测细胞的迁移能力；Western blot实验用于验证MMP14蛋白在恶性转化16HBE细胞中的表达情况，并检测ERK1/2及其磷酸化蛋白的表达水平；采用实时荧光定量PCR(qPCR)法检测两组细胞中MMP14和ERK通路关键信号分子ERK1、ERK2的mRNA表达水平。结果：GMA处理组细胞在软琼脂中形成的集落数显著多于DMSO对照组(P<0.05)。细胞划痕实验和Transwell细胞迁移实验结果显示，GMA诱导的恶性转化16HBE细胞的整体和个体迁移能力均显著大于DMSO对照组细胞(P<0.05)。与DMSO对照组相比，恶性转化16HBE细胞中MMP14 mRNA和蛋白的表达水平升高；p-ERK1/2蛋白和ERK1 mRNA的表达水平亦升高，差异具有统计学意义(P<0.05)，而ERK2 mRNA的表达水平差异无统计学意义(P>0.05)。结论：GMA诱导16HBE细胞的恶性转化过程可能与ERK/MMP14信号通路的激活相关，本研究结果为GMA诱导16HBE细胞恶性转化的致癌机制研究提供了新的线索。

关键词: 甲基丙烯酸缩水甘油酯, 16HBE细胞, 基质金属蛋白酶14, 细胞外调节蛋白激酶1/2

Abstract: OBJECTIVE：To investigate whether glycidyl methacrylate (GMA) would affect malignant transformation of human bronchial epithelial (16HBE) cells through the ERK/MMP14 signaling pathway，and to investigate its molecular mechanism for malignant transformation of the 16HBE cells. METHODS：16HBE cells which were repeatedly exposed to 8 μg/mL GMA were used as the GMA treatment group，and the cells treated with the same volume of dimethyl sulfoxide (DMSO) as the solvent control group. At the 40th passage of the 16HBE cells，they were harvested，and the degree of malignant transformation was identified using the soft agar clone formation assay. The wound healing and the transwell cell migration assays were used to detect the migration ability of cells. Western blot was used to verify expression of the MMP14 protein，and levels of ERK1/2 and its phosphorylated proteins. Quantitative real-time PCR (qPCR) was used to detect mRNA expression levels of MMP14 and of the key signal molecules of the ERK pathway：ERK1 and ERK2. RESULTS：The number of colonies formed by cells in the GMA-treated group in soft agar was greater than that in the DMSO-treated group (P<0.05). Results from the wound healing and the transwell cell migration assays showed that the migration ability of the GMA-treated 16HBE cells were significantly higher than those of the DMSO-treated control cells (P<0.05). Compared with the control cells，expression levels of MMP14 mRNA and protein in 16HBE cells were increased. The expression levels of p-ERK1/2 protein and ERK1 mRNA were also increased，and the differences were statistically significant (P<0.05)，while there was no significant difference in ERK2 mRNA expression (P<0.05). CONCLUSION：The process of GMA-induced malignant transformation of 16HBE cells was associated with activation of the ERK/MMP14 signaling pathway. Our results are helpful for a better understanding of mechanism in GMA-induced malignant transformation of 16HBE cells.

Key words: glycidyl methacrylate, 16HBE cells, MMP14, ERK1/2

中图分类号:

R730.2

崔旭芳, 王全凯, 金惠萍, 李昕苇, 顾轶婷, 乌瀚宝栎尔, 康同影, 许建宁. 甲基丙烯酸缩水甘油酯通过ERK/MMP14信号通路影响16HBE细胞恶性转化的研究[J]. 癌变·畸变·突变, 2024, 36(4): 261-267.

CUI Xufang, WANG Quankai, JIN Huiping, LI Xinwei, GU Yiting, WUHAN Baolier, KANG Tongying, XU Jianning. Glycidyl methacrylate affects malignant transformation of 16HBE cells through the ERK/MMP14 signaling pathway[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2024, 36(4): 261-267.

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甲基丙烯酸缩水甘油酯通过ERK/MMP14信号通路影响16HBE细胞恶性转化的研究

Glycidyl methacrylate affects malignant transformation of 16HBE cells through the ERK/MMP14 signaling pathway

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