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. 2021 Feb 1;39(1):81-87.
doi: 10.7518/hxkq.2021.01.012.

Effect of chromodomain helicase/ATPase DNA binding protein 1-like gene on the invasion and metastasis of tongue squamous cell carcinoma CAL27 cells

[Article in English, Chinese]
Kai-Li Hu  1 , Xin Fan  2 , Wen-Ting Hu  3 , Hong-Li Li  4 , Qing-Hua Tang  1 , Xue-Hui Sun  3
Affiliations

Effect of chromodomain helicase/ATPase DNA binding protein 1-like gene on the invasion and metastasis of tongue squamous cell carcinoma CAL27 cells

[Article in English, Chinese]
Kai-Li Hu et al. Hua Xi Kou Qiang Yi Xue Za Zhi. .

Abstract
in English, Chinese

Objectives: A study was conducted to investigate the molecular mechanism of chromodomain helicase/ATPase DNA binding protein 1-like gene (CHD1L) influencing the invasion and metastasis of tongue squamous cell carcinoma and to provide a new target for clinical inhibition of invasion and metastasis of tongue squamous cell carcinoma.

Methods: Ualcan website was used to analyze the expression of CHD1L in normal epithelial tissue and primary head and neck squamous cell carcinoma and to analyze the effect of lymph node metastasis on the expression of CHD1L in tissues with head and neck squamous cell carcinoma. The relationship between CHD1L expression and the survival rate of patients with head and neck squamous cell carcinoma was tested by the GEPIA website. Western blot was used to quantify the levels of CHD1L protein in human tongue squamous cell carcinoma CAL27 and immortalized human skin keratinocyte cell HaCaT. After knocking down CAL27 in human tongue squamous cell carcinoma cells with an RNA interference plasmid, the cells were designated as SiCHD1L/CAL27 and Scr/CAL27. Western blot was utilized to detect the expression of CHD1L in each group of cells. The change in CAL27 cell proliferation ability was tested by EdU proliferation test after CHD1L knockdown. The change of cell migration ability of each group cells was tested through the wound healing assay. Western blot was used to detect epithelial-mesenchymal transition (EMT) marker E-cadherin and Vimentin protein expression levels.

Results: Ualcan database showed that the expression of CHD1L in primary head and neck squamous cell carcinoma tissues was higher than in normal epithelial tissues and in head and neck squamous cell carcinoma tissues with lymph node metastasis. GEPIA website analysis showed that the overall survival rate of patients with head and neck squamous cell carcinoma with high expression of CHD1L was significantly lower than that of patients with low expression. Western blot results showed that CHD1L expression in human tongue squamous carcinoma cells CAL27 was higher than that of human normal skin cells HaCaT. CHD1L expression in SiCHD1L/CAL27 cells was much lower than that in Scr/CAL27 cells. Results of EdU proliferation experiments showed the significant reduction in the cell proliferation ability of the SiCHD1L/CAL27 cells. Results of the wound healing experiments showed the reduction in the migration capacity of the SiCHD1L/CAL27 cells. The expression of E-cadherin increased, whereas that of Vimentin decreased, in SiCHD1L/CAL27 cells.

Conclusions: CHD1L promoted the EMT, proliferation, migration, and invasion ability of tongue squamous cell carcinoma cells.

目的: 探讨染色质解旋酶DNA结合蛋白（CHD1L）影响舌鳞状细胞癌侵袭和转移的分子机制，为临床抑制舌鳞状细胞癌的侵袭和转移提供新的靶点。方法: 运用Ualcan网站分析CHD1L在正常上皮组织和原发性头颈鳞状细胞癌组织中的表达情况，分析有无淋巴结转移对头颈鳞状细胞癌组织中CHD1L表达情况的影响；通过GEPIA网站分析CHD1L在头颈鳞状细胞癌中的表达与生存率的关系；蛋白质印迹法检测人舌鳞状细胞癌细胞CAL27和人正常皮肤细胞HaCaT中CHD1L蛋白的表达水平；用RNA干扰质粒敲低人舌鳞状细胞癌细胞CAL27中的CHD1L后，将细胞分别命名为SiCHD1L/CAL27及Scr/CAL27；蛋白质印迹法检测CHD1L在每组细胞中的表达情况；EdU增殖实验检测每组细胞增殖能力的改变；伤口愈合实验检测每组细胞迁移能力的改变；蛋白质印迹法检测上皮间质转化（EMT）标志物上皮钙黏着蛋白和波形蛋白的表达水平。结果: Ualcan数据库显示：CHD1L在原发头颈鳞状细胞癌组织中表达高于正常上皮组织中的表达，在出现淋巴结转移的头颈鳞状细胞癌组织中表达更高。GEPIA网站分析显示：CHD1L表达量高的头颈鳞状细胞癌患者总生存率较表达量低的患者明显降低。蛋白质印迹结果显示：人舌鳞状细胞癌细胞CAL27中CHD1L表达高于人正常皮肤细胞HaCaT中CHD1L的表达，敲低CHD1L以后CAL27细胞中CHD1L的表达量明显低于Scr/CAL27组细胞。EdU增殖实验结果显示：SiCHD1L/CAL27组细胞增殖能力明显降低。伤口愈合实验显示：SiCHD1L/CAL27组迁移能力降低，SiCHD1L/CAL27组细胞上皮钙黏着蛋白表达量增加而波形蛋白表达量降低。结论: CHD1L促进舌鳞状细胞癌细胞的EMT，促进舌鳞状细胞癌细胞的增殖、迁移侵袭能力。.

Keywords: chromodomain helicase/ATPase DNA gene; epithelial-mesenchymal transition; invasion; proliferation; tongue squamous cell carcinoma.

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Conflict of interest statement

利益冲突声明：作者声明本文无利益冲突。

Figures

图 1
图 1. CHD1L在头颈鳞状细胞癌组织中的表达及与患者生存分析
Fig 1 Expression of CHD1L in squamous cell carcinoma of the head and neck and its survival analysis A：Ualcan数据库中CHD1L在正常上皮组织和头颈鳞状细胞癌组织中的表达情况；B：Ualcan数据库中CHD1L在不同淋巴结转移数量的头颈鳞状细胞癌组织中的表达情况；C：GEPIA数据库中CHD1L的表达与头颈鳞状细胞癌患者生存情况的分析图。
图 2
图 2. CHD1L在各组细胞中的表达
Fig 2 Expression of CHD1L in indicated cells 左：CHD1L在人舌鳞状细胞癌细胞CAL27和人正常皮肤细胞HaCaT中的表达情况；右：CHD1L在Scr/CAL27细胞和SiCHD1L/CAL27细胞中的表达情况；上：蛋白质印迹检测图像；下：CHD1L蛋白的相对表达量。*P<0.05。
图 3
图 3. CHD1L对Scr/CAL27和SiCHD1L/CAL27增殖能力的影响
Fig 3 Effect of CHD1L on the proliferation ability of Scr/CAL27 and SiCHD1L/CAL27 cells 左：Scr/CAL27细胞和SiCHD1L/CAL27细胞的共聚焦显微镜图像；右：Scr/CAL27细胞和SiCHD1L/CAL27细胞的增殖情况。*P<0.05。
图 4
图 4. CHD1L对Scr/CAL27和SiCHD1L/CAL27迁移能力的影响
Fig 4 Effect of CHD1L on the migration ability of Scr/CAL27 and SiCHD1L/CAL27 cells 左：Scr/CAL27细胞和SiCHD1L/CAL27细胞的伤口愈合实验图像；右：Scr/CAL27细胞和SiCHD1L/CAL27细胞的相对迁移率。*P<0.05。
图 5
图 5. EMT相关蛋白在Scr/CAL27和SiCHD1L/CAL27的表达情况
Fig 5 Expression of EMT related proteins in Scr/CAL27 and SiCHD1L/CAL27 cells 左：上皮钙黏着蛋白和波形蛋白的蛋白质印迹图像；右：上皮钙黏着蛋白和波形蛋白的相对表达量。*P<0.05。
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