ATIP1和ATIP3a对口腔恶性肿瘤细胞增殖和凋亡的影响

doi:10.19438/j.cjoms.2021.05.006

摘要/Abstract

摘要： 目的探讨过表达MTUS1/ATIPs对口腔恶性肿瘤细胞增殖及凋亡的影响。方法应用RT-PCR检测舌鳞癌（TSCC）细胞UM1、腺样囊性癌组织（ACC）和唾液腺腺样囊性癌细胞（SACC-83）中MTUS1/ATIP表达水平及亚型分布。应用含ATIP1和ATIP3a片段的质粒转染口腔恶性肿瘤细胞UM1和SACC-83,48 h后以MTT检测细胞的增殖能力。采用流式细胞仪技术和细胞免疫荧光技术检测细胞周期和细胞凋亡率,Western免疫印迹检测细胞中ATIP1、ATIP3a、ERK1/2、pERK1/2、Caspase3的表达水平。采用SPSS 19.0软件包对数据进行统计学处理。结果 RT-PCR结果显示,ATIP1、ATIP3a、ATIP3b是MTUS1的主要亚型,在口腔恶性肿瘤细胞（TSCC细胞系UM1、ACC组织和SACC-83细胞）中表达显著降低。转染MTUS1/ATIP1和MTUS1/ATIP3a后,舌鳞癌细胞的增殖能力受到显著抑制。转染ATIP1后,UM1细胞的抑制率分别为38.8%（24 h）和57.3%（48 h）;转染ATIP3a后,UM1细胞的抑制率分别为48.1%（24 h）和56.5%（48 h）;同时转染ATIP1和ATIP3a后,UM1细胞的抑制率分别为93.5%（24 h）和88.8%（48 h）。同样,在SACC-83细胞中也出现类似效果（P<0.05）。细胞周期分析发现,高表达MTUS1/ATIP1后,UM1及SACC-83细胞阻滞在G1/G0期;而ATIP3a则诱导细胞阻滞在G2/M期（P<0.05）。Western免疫印迹检测显示,转染MTUS1/ATIP1和MTUS1/ATIP3a后,ERK表达升高,磷酸化ERK表达下降,Caspase 3表达升高。结论 MTUS1/ATIP是一个潜在的抑癌基因,可通过ERK途径,调控口腔恶性肿瘤细胞增殖并诱导细胞凋亡。

关键词: 口腔恶性肿瘤, MTUS1/ATIP1, MTUS1/ATIP3a, 增殖, 凋亡, ERK

Abstract: PURPOSE: To investigate the role of MTUS1/ATIP in suppression of proliferation and induce apoptosis of oral malignant tumor cells. METHODS: The expression level of MTUS1/ATIP in oral malignant tumor cells (UM1, SACC-83) was detected. Oral malignant tumor cells were transfected with ATIP1 and ATIP3a vector for 48 h. Proliferation analysis was performed with MTT assay. Flow cytometry and immunofluorescence technique were employed to measure cell cycle and apoptosis of oral malignant tumor cells. The expression level of proteins ATIP1, ATIP3a, Caspase3, ERK1/2, pERK1/2 in oral malignant tumor cells transfected with ATIP1 and ATIP3a was detected by Western blotting. The data was analyzed using SPSS 19.0 software package. RESULTS: ATIP1, ATIP3a and ATIP3b were the major isoforms produced by MTUS1 gene and significantly decreased in oral malignant tumor cells [TSCC cell line UM1, parotid adenoid cystic carcinoma (ACC) tissue and cell line SACC-83]. Restoration of MTUS1 (ATIP1 and/or ATIP3a) expression induced anti-proliferative activities against oral malignant tumor cells (UM1 and SACC-83), with a cooperative effect between ATIP1 and ATIP3a. In TSCC cells, the inhibition rate of ATIP1 was approximately 38.8% (24 h) and 57.3% (48 h), the inhibition rate of ATIP3a was approximately 48.1% (24 h) and 56.5% (48 h); however, the inhibition rate increased to 93.5% (24 h) and 88.8% (48 h) with combined action of ATIP1 and ATIP3a in TSCC cell lines (P<0.05). A similar effect was observed in adenoid cystic carcinoma (ACC) cells (P<0.05). Over-expression of ATIP1 led to G1/G0 cell cycle arrest, whereas ATIP3a caused G2/M arrest (P<0.05). Both ATIP1 and ATIP3a induced apoptosis and inhibited phosphorylation of ERK1/2 (extracellular-regulated kinase). CONCLUSIONS: The results suggest that MTUS1 is a promising anticancer agent for oral malignant tumor and has apoptosis-inducing activities via ERK1/2 pathway.

Key words: Oral malignant tumor, MTUS1/ATIP, Proliferation, Apoptosis, ERK

中图分类号:

R739.8

赵婷婷, 冯艳青, 何倩婷, 莫云龙, 董恩恩, 王安训. ATIP1和ATIP3a对口腔恶性肿瘤细胞增殖和凋亡的影响[J]. 中国口腔颌面外科杂志, 2021, 19(5): 417-423.

ZHAO Ting-ting, FENG Yan-qing, HE Qian-ting, MO Yun-long, DONG En-en, WANG An-xun. ATIP1 and ATIP3a, two isoforms of MTUS1/ATIP, suppressed proliferation and induced apoptosis in oral malignant tumor cells[J]. China J Oral Maxillofac Surg, 2021, 19(5): 417-423.

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