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

Abstract

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

CLC Number:

R730.2

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

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