下调SETDB1通过SOX7甲基化抑制口腔癌上皮-间充质转化、侵袭和转移的机制探讨

doi:10.19439/j.sjos.2024.01.005

摘要/Abstract

摘要： 目的: 探讨SETDB1通过SOX7甲基化抑制口腔癌上皮-间充质转化（epithelial mesenchymal transition,EMT）、迁移和侵袭机制。方法: 应用qRT-PCR和Western印迹法检测口腔癌KB细胞和口腔黏膜上皮ATCC细胞中SETDB1 和SOX7 mRNA和蛋白表达水平。构建SETDB1 si-RNA,以脂质体介导法转染口腔癌KB细胞。siRNA-SETDB1为实验组（si-S）,siRNA空载体为阴性对照组（si-N）,未转染KB细胞为空白对照组（NC）。qRT-PCR和Western印迹法检测SETDB1 mRNA和蛋白表达水平,验证转染效果;焦磷酸测序检测SOX7甲基化水平。Western印迹法检测N-钙黏蛋白（N-cadherin）、Vimentin、β-catenin和Slug蛋白表达,MTT法检测细胞存活能力,流式细胞术检测细胞凋亡,划痕愈合实验检测细胞迁移能力,Transwell小室实验检测细胞侵袭能力。采用SPSS 21.0软件包进行统计学分析。结果: Rt-qPCR和Western印迹法检测结果显示,si-S组SETDB1 mRNA和蛋白表达量显著下降（P<0.05）。焦磷酸测序检测结果显示,下调SETDB1可显著降低SOX7甲基化率,增加SOX7蛋白表达。Western印迹法检测结果显示,下调SETDB1可显著抑制口腔癌KB细胞中EMT相关蛋白N-cadherin、Vimentin、β-catenin和Slug的表达（P<0.05）。细胞功能学结果显示,下调SETDB1可显著抑制KB细胞的存活、迁移和侵袭能力。结论: 下调SETDB1可通过调控SOX7甲基化水平,抑制口腔癌细胞EMT、迁移和侵袭,为口腔癌临床诊断和治疗提供新思路和靶点。

关键词: SETDB1, 甲基转移酶, SOX7, 甲基化, 口腔癌

Abstract: PURPOSE: To explore the mechanism of SETDB1 inhibiting epithelial mesenchymal transition (EMT),migration and invasion in oral cancer via SOX 7 methylation. METHODS: SETDB1 and SOX7 mRNA and protein expression levels in KB cells of oral cancer and oral mucosal epithelial ATCC cells were determined by qRT-PCR and Western blot (WB). SETDB1 si-RNA was structured, then transfect into KB cells of oral cancer by liposome-mediated method. siRNA-SETDB1 was the experimental group (si-S), siRNA empty vector was the negative control group (si-N), and untransfected KB cells were the blank control group(NC). SETDB1 mRNA and protein expression levels were detected by qRT-PCR and Western blot(WB), to verify the transfection effect. The methylation levels of SOX7 were determined by pyrosequencing. The expression of N-cadherin, Vimentin, β-catenin, and Slug proteins was detected by WB. Cell viability was measured by MTT assay, migration ability was tested by scratch healing assay, and invasion ability was tested by Transwell chamber assay. Statistical analysis was performed with SPSS 21.0 software package. RESULTS: The results of Rt-qPCR and WB showed that the SETDB1 mRNA and protein expression decreased significantly in si-S group(P<0.05). Pyrosequencing test results showed that the regulation of SETDB1 could significantly reduce the SOX7 methylation rate and increased the SOX7 protein expression. WB results showed that knockdown of SETDB1 significantly inhibited the expression of EMT-related proteins N-cadherin, Vimentin, β-catenin and Slug in oral cancer KB cells (P<0.05). The results of cell functology experiments showed that knockdown of SETDB1 could significantly inhibit survival, migration and invasion of KB cells. CONCLUSIONS: Downregulation of SETDB1 could suppress EMT, migration and invasion of oral cancer cells by regulating SOX7 methylation level, providing new ideas and targets for the diagnosis and treatment of oral cancer.

Key words: SETDB1, Methyltransferase, SOX 7, Methylation, Oral cancer

中图分类号:

R739.8

郝妍, 白相宇, 霍峰, 陈喜波. 下调SETDB1通过SOX7甲基化抑制口腔癌上皮-间充质转化、侵袭和转移的机制探讨[J]. 上海口腔医学, 2024, 33(1): 30-35.

HAO Yan, BAI Xiang-yu, HUO Feng, CHEN Xi-bo. Downregulation of SETDB1 suppresses epithelial mesenchymal transition and invasion and metastasis in oral cancer via SOX7 methylation[J]. Shanghai Journal of Stomatology, 2024, 33(1): 30-35.

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