NUDT1促进口腔鳞状细胞癌细胞增殖的体外实验研究

doi:10.12439/kqhm.1005-4979.2025.01.004

摘要/Abstract

摘要：

目的： 观察Nudix水解酶1（Nudix hydrolase 1，NUDT1）对口腔鳞状细胞癌（oral squamous cell carcinoma，OSCC）细胞增殖的作用，并初步探讨其作用机制。方法： 应用免疫组织化学法对30例OSCC患者的肿瘤组织和10例正常口腔黏膜组织石蜡切片进行检测。蛋白质印迹法（Western blotting）检测NUDT1在OSCC细胞和正常口腔黏膜上皮细胞中的表达。用小干扰RNA（small interfering RNA，siRNA）干扰法下调NUDT1的表达，CCK8实验检测在OSCC细胞（CAL27细胞系和HN30细胞系）中沉默NUDT1对细胞增殖的影响，用凋亡试剂盒检测沉默NUDT1对细胞凋亡的影响。结果：免疫组织化学检测结果显示，NUDT1在OSCC组织中的表达高于其在正常口腔黏膜组织中，Western blotting检测结果显示，与正常口腔黏膜上皮细胞相比，NUDT1在OSCC细胞CAL27和HN30中表达升高（P<0.01）。抑制NUDT1的基因表达能明显抑制OSCC细胞（CAL27和HN30细胞系）的增殖能力（P<0.05），并且促进OSCC细胞的凋亡。结论：与正常口腔黏膜组织相比，NUDT1在OSCC组织中表达升高，下调NUDT1对于抑制OSCC细胞的增殖和促进OSCC细胞凋亡具有重要意义。

关键词: 口腔鳞状细胞癌, Nudix水解酶1, 增殖, 凋亡

Abstract:

Objective: To investigate the effect of Nudix hydrolase 1 (NUDT1) on the proliferation of oral squamous cell carcinoma (OSCC) cells, and its mechanism of function was tentatively explored.Methods: Immunohistochemistry was applied to examine paraffin sections of 30 patient tissues with OSCC and 10 normal oral mucosa tissues. The expression of NUDT1 in OSCC cells and normal oral mucosal epithelial cells was detected by Western blotting. In addition, the expression of NUDT1 was down-regulated by small interfering RNA (siRNA) interference. The effect of NUDT1 silencing on cell proliferation in OSCC cell lines CAL27 and HN30 was examined by CCK8 assay, and the effect of NUDT1 silencing on cell apoptosis was analyzed by immunofluorescence through an apoptosis kit.Results: Immunohistochemistry assay showed that NUDT1 expression was higher in OSCC tissues than that in normal oral mucosa tissues, and Western blotting assay revealed that NUDT1 expression was elevated in OSCC cells (CAL27 and HN30 cells) compared with that in normal oral mucosal epithelial cells (P<0.01). Inhibition of NUDT1 gene expression significantly inhibited the proliferation ability of OSCC cells (P<0.05), and promoted apoptosis of OSCC cells.Conclusion: NUDT1 expression was elevated in OSCC tissues, compared to that in normal oral mucosa tissues. Downregulation of NUDT1 was important for inhibiting the proliferation of OSCC cells and promoting apoptosis of OSCC cells.

Key words: oral squamous cell carcinoma, Nudix hydrolase 1, proliferation, apoptosis

中图分类号:

R782

郭浩兵, 鲁莉莉, 郝雅翠, 赵博文, 高艺涵, 朴松林. NUDT1促进口腔鳞状细胞癌细胞增殖的体外实验研究[J]. 《口腔颌面外科杂志》, 2025, 35(1): 21-27.

GUO Haobing, LU Lili, HAO Yacui, ZHAO Bowen, GAO Yihan, PIAO Songlin. In vitro experimental study of NUDT1 promoting the proliferation of oral squamous cell carcinoma cells[J]. 《Journal of Oral and Maxillofacial Surgery》, 2025, 35(1): 21-27.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2025/V35/I1/21

图/表 5

图1 正常组织与OSCC组织中NUDT1的表达水平 A.正常组织免疫组织化学染色图(×400)；B.高分化OSCC组织免疫组织化学染色图(×400)；C.低分化OSCC组织免疫组织化学染色图(×400)；D. NUDT1在不同病理等级组织中的比较，***表示P<0.001，与正常组织比较，##表示P<0.01，与高分化OSCC组织比较。

Figure 1 Expression levels of NUDT1 in normal tissues vs OSCC tissues

图2 NUDT1在HOMEC、CAL27与HN30细胞中的表达 A. NUDT1与GAPDH的蛋白表达条带；B.各组间NUDT1蛋白相对表达量比较，**表示P<0.01，与HOMEC组比较。

Figure 2 Expression of NUDT1 in HOMEC vs CAL27 and HN30 cells

图3 NUDT1 siRNA的转染效率 A.蛋白表达条带；B. CAL27细胞NUDT1蛋白表达定量分析；C. HN30细胞NUDT1蛋白表达定量分析。**表示P<0.01，***表示P<0.001，与Si-NUDT1组比较。

Figure 3 Transfection efficiency of NUDT1 siRNA

图4 抑制NUDT1基因表达对OSCC细胞系增殖的影响 A.抑制NUDT1基因表达影响CAL27细胞增殖；B.抑制NUDT1基因表达影响HN30细胞增殖。*表示P<0.05，**表示P<0.01，与Si-NUDT1组比较。

Figure 4 Effects of inhibition of NUDT1 gene expression on the proliferation of OSCC cell lines

图5 抑制NUDT1基因表达促进OSCC细胞系凋亡(×200) A. CAL27细胞凋亡（左列）与坏死（右列）结果；B. HN30细胞凋亡（左列）与坏死（右列）结果。正常细胞显示为暗蓝色，凋亡细胞显示为亮蓝色，坏死细胞显示为亮红色。

Figure 5 Inhibition of NUDT1 gene expression promotes apoptosis in OSCC cell lines (×200)

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