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抑制乙酰辅酶A羧化酶1促进食管癌细胞迁移

[Knockdown of ACC1 promotes migration of esophageal cancer cell].

作者信息

Qian H, Gu C W, Liu Y Z, Zhao B S

机构信息

Department of Thoracic Surgery, the First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, China.

Life Science Research Center, the First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2023 Jun 23;45(6):482-489. doi: 10.3760/cma.j.cn112152-20210517-00383.

Abstract

To investigate the effect of acetyl-CoA carboxylase 1 (ACC1) knockdown on the migration of esophageal squamous cell carcinoma (ESCC) KYSE-450 cell and underlying mechanism. Lentiviral transfection was conducted to establish sh-NC control cell and ACC1 knocking down cell (sh-ACC1). Human siRNA HSP27 and control were transfected by Lipo2000 to get si-HSP27 and si-NC. The selective acetyltransferase P300/CBP inhibitor C646 was used to inhibit histone acetylation and DMSO was used as vehicle control. Transwell assay was performed to detect cell migration. The expression of HSP27 mRNA was examined by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) and the expressions of ACC1, H3K9ac, HSP27 and epithelial-mesenchymal transition-related proteins E-cadherin and Vimentin were detected by western blot. The expression level of ACC1 in sh-NC group was higher than that in sh-ACC1 group (<0.01). The number of cell migration in sh-NC group was (159.00±24.38), lower than (361.80±26.81) in sh-ACC1 group (<0.01). The protein expression levels of E-cadherin and Vimentin in sh-NC group were statistically significant compared with sh-AAC1 group (<0.05). The migrated cell number in sh-NC+ si-NC group was (189.20±16.02), lower than (371.60±38.40) in sh-ACC1+ si-NC group (<0.01). The migrated cell number in sh-NC+ si-NC group was higher than that in sh-NC+ si-HSP27 group (152.40±24.30, <0.01), and the migrated cell number in sh-ACC1+ si-NC group was higher than that in sh-ACC1+ si-HSP27 group (<0.01). The protein expression levels of E-cadherin and Vimentin in sh-NC+ si-NC group were significantly different from those in sh-ACC1+ si-NC and sh-NC+ si-HSP27 groups (<0.01). The protein expression levels of E-cadherin and Vimentin in sh-ACC1+ si-NC group were significantly different from those in sh-ACC1+ si-HSP27 group (<0.01). After 24 h treatment with C646 at 20 μmmo/L, the migrated cell number in sh-NC+ DMSO group was (190.80±11.95), lower than (395.80±17.10) in sh-ACC1+ DMSO group (<0.01). The migrated cell number in sh-NC+ DMSO group was lower than that in sh-NC+ C646 group (256.20±23.32, <0.01). The migrated cell number in sh-ACC1+ DMSO group was higher than that in sh-ACC1+ C646 group (87.80±11.23, <0.01). The protein expressions of H3K9ac, HSP27, E-cadherin and Vimentin in sh-NC+ DMSO group were significantly different from those in sh-ACC1+ DMSO group and sh-NC+ C646 group (<0.01). The protein expression levels of H3K9ac, HSP27, E-cadherin and Vimentin in sh-ACC1+ DMSO group were significantly different from those in sh-ACC1+ C646 group (<0.01). Knockdown of ACC1 promotes the migration of KYSE-450 cell by up-regulating HSP27 and increasing histone acetylation.

摘要

探讨乙酰辅酶A羧化酶1(ACC1)基因敲低对食管鳞状细胞癌(ESCC)KYSE-450细胞迁移的影响及其潜在机制。通过慢病毒转染建立sh-NC对照细胞和ACC1基因敲低细胞(sh-ACC1)。采用脂质体2000转染人源小干扰RNA(siRNA)HSP27和对照,获得si-HSP27和si-NC。使用选择性乙酰转移酶P300/CBP抑制剂C646抑制组蛋白乙酰化,以二甲基亚砜(DMSO)作为溶剂对照。采用Transwell实验检测细胞迁移。通过逆转录-定量实时聚合酶链反应(RT-qPCR)检测HSP27 mRNA的表达,采用蛋白质免疫印迹法检测ACC1、H3K9ac、HSP27以及上皮-间质转化相关蛋白E-钙黏蛋白和波形蛋白的表达。sh-NC组ACC1的表达水平高于sh-ACC1组(<0.01)。sh-NC组细胞迁移数为(159.00±24.38),低于sh-ACC1组的(361.80±26.81)(<0.01)。sh-NC组E-钙黏蛋白和波形蛋白的蛋白质表达水平与sh-AAC1组相比差异有统计学意义(<0.05)。sh-NC+si-NC组迁移细胞数为(189.20±16.02),低于sh-ACC1+si-NC组的(371.60±38.40)(<0.01)。sh-NC+si-NC组迁移细胞数高于sh-NC+si-HSP27组(152.40±24.30,<0.01),sh-ACC1+si-NC组迁移细胞数高于sh-ACC1+si-HSP27组(<0.01)。sh-NC+si-NC组E-钙黏蛋白和波形蛋白的蛋白质表达水平与sh-ACC1+si-NC组和sh-NC+si-HSP27组相比差异有统计学意义(<0.01)。sh-ACC1+si-NC组E-钙黏蛋白和波形蛋白的蛋白质表达水平与sh-ACC1+si-HSP27组相比差异有统计学意义(<0.01)。用20 μmmol/L C646处理24小时后,sh-NC+DMSO组迁移细胞数为(190.80±11.95),低于sh-ACC1+DMSO组的(395.80±17.10)(<0.01)。sh-NC+DMSO组迁移细胞数低于sh-NC+C646组(256.20±23.32,<0.01)。sh-ACC1+DMSO组迁移细胞数高于sh-ACC1+C646组(87.80±11.23,<0.01)。sh-NC+DMSO组H3K9ac、HSP27、E-钙黏蛋白和波形蛋白的蛋白质表达与sh-ACC1+DMSO组和sh-NC+C646组相比差异有统计学意义(<0.01)。sh-ACC1+DMSO组H3K9ac、HSP27、E-钙黏蛋白和波形蛋白的蛋白质表达水平与sh-ACC1+C646组相比差异有统计学意义(<0.01)。ACC1基因敲低通过上调HSP27和增加组蛋白乙酰化促进KYSE-450细胞迁移。

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