c-Kit蛋白在甲基丙烯酸缩水甘油酯诱导的恶性转化前期16HBE细胞中的作用机制

doi:10.3969/j.issn.1004-616x.2025.06.001

癌变·畸变·突变 ›› 2025, Vol. 37 ›› Issue (6): 425-431.doi: 10.3969/j.issn.1004-616x.2025.06.001

• 论著 •

c-Kit蛋白在甲基丙烯酸缩水甘油酯诱导的恶性转化前期16HBE细胞中的作用机制

崔旭芳^1,2, 李昕苇^1,2, 周倩^1,2, 张岩^1,2, 许建宁^1,2, 王全凯^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:2025-08-07 修回日期:2025-09-15 发布日期:2025-12-06
通讯作者: 王全凯，E-mail：kylewang@sina.com
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)

Mechanism of c-Kit protein in premalignant transformation of 16HBE cells induced by glycidyl methacrylate

CUI Xufang^1,2, LI Xinwei^1,2, ZHOU Qian^1,2, ZHANG Yan^1,2, XU Jianning^1,2, WANG Quankai^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:2025-08-07 Revised:2025-09-15 Published:2025-12-06

摘要/Abstract

摘要： 目的：研究甲基丙烯酸缩水甘油酯(GMA)诱导的恶性转化前期16HBE细胞中c-Kit蛋白的作用机制。方法：使用8 μg/mL的GMA对16HBE细胞进行3次重复染毒，同体积二甲基亚砜(DMSO)处理细胞作为对照组，收获第10代(P10)的两组细胞。分别采用逆转录实时荧光定量PCR(RT-qPCR)和Western blot实验检测两组细胞中c-Kit mRNA和蛋白的表达水平。进一步使用5和20 μmol/L c-Kit蛋白特异性抑制剂ISCK03作用GMA处理组细胞24 h后，采用Western blot实验检测通路相关蛋白PI3K、Akt及其磷酸化水平，细胞周期相关蛋白Cyclin D1、Cyclin B1和凋亡蛋白Caspase-3的表达水平；采用流式细胞术检测各组细胞周期和细胞凋亡率的变化。结果：与DMSO对照组相比，P10代GMA处理组细胞中c-Kit mRNA和蛋白表达水平均显著降低(P<0.01)。与GMA处理组相比，使用20 μmol/L ISCK03抑制c-Kit蛋白表达后，PI3K和Akt蛋白的磷酸化水平降低、Cyclin B1和Caspase-3蛋白的表达水平下降、Cyclin D1蛋白的表达水平升高、细胞凋亡率降低，差异均具有统计学意义(均为P<0.05)。结论：c-Kit蛋白可能通过抑制PI3K/Akt信号通路调控细胞周期和细胞凋亡过程，在细胞恶性转化前期发挥重要作用。本研究为GMA潜在的致癌机制研究提供了参考。

关键词: 甲基丙烯酸缩水甘油酯, 16HBE细胞, 恶性转化前期, c-Kit蛋白

Abstract: OBJECTIVE：To investigate the mechanistic role of downregulating the differentially expressed protein c-Kit in glycidyl methacrylate (GMA)-induced premalignant transformation of 16HBE cells. METHODS：Three replicates of 16HBE cells were treated with 8 μg/mL GMA，and the same volume of dimethyl sulfoxide (DMSO) treatment served as a control group. The GMA- and DMSO-treated cells were passaged to the 10^th generation (P10) and then harvested. Real-time quantitative PCR (RT-qPCR) and Western blot experiments were performed to detect expression levels of c-Kit mRNA and protein in the two groups of cells. Following 24-hour treatment of GMA-treated cells with 5 and 20 μmol/L of the c-Kit-specific inhibitor ISCK03，western blot assay was performed to detect pathway-related proteins including PI3K，Akt，and their phosphorylation levels，as well as the expression levels of cycle-related proteins Cyclin D1 and Cyclin B1，and the apoptotic protein Caspase-3. Flow cytometry was employed to assess cell cycle progression and apoptotic changes across all groups. RESULTS：Compared with the DMSO control group，both c-Kit mRNA and protein expression levels were reduced in P10 cells treated with GMA (P<0.01). Following inhibition of c-Kit protein expression using 5 and 20 μmol/L ISCK03，compared to the GMA-treated group，the phosphorylation levels of PI3K and Akt proteins decreased，the expression levels of Cyclin B1 and Caspase-3 proteins declined，while the expression level of Cyclin D1 protein increased. Furthermore，cell cycle progression was impeded，and apoptosis rate was decreased，with all differences being statistically significant (all P<0.05). CONCLUSION：C-Kit protein may play an important role in the premalignant transformation of cells by regulating the cell cycle and apoptotic processes through inhibition of the PI3K/Akt signaling pathway. This study provides a reference for the mechanistic study of the potential carcinogenicity of GMA.

Key words: glycidyl methacrylate, 16HBE cells, premalignant transformation, c-Kit protein

中图分类号:

R730.2

崔旭芳, 李昕苇, 周倩, 张岩, 许建宁, 王全凯. c-Kit蛋白在甲基丙烯酸缩水甘油酯诱导的恶性转化前期16HBE细胞中的作用机制[J]. 癌变·畸变·突变, 2025, 37(6): 425-431.

CUI Xufang, LI Xinwei, ZHOU Qian, ZHANG Yan, XU Jianning, WANG Quankai. Mechanism of c-Kit protein in premalignant transformation of 16HBE cells induced by glycidyl methacrylate[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2025, 37(6): 425-431.

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c-Kit蛋白在甲基丙烯酸缩水甘油酯诱导的恶性转化前期16HBE细胞中的作用机制

Mechanism of c-Kit protein in premalignant transformation of 16HBE cells induced by glycidyl methacrylate

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