首页|敲低Sec31A对头颈鳞状细胞癌恶性表型的影响

敲低Sec31A对头颈鳞状细胞癌恶性表型的影响

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目的 探讨敲低Sec31A对头颈鳞状细胞癌(head and neck squamous cell carcinoma,HNSCC)恶性表型的影响及可能机制.方法 从TCGA数据库中获得HNSCC组织与癌旁组织的转录组测序数据并比较Sec31A的表达水平,免疫组化染色分析Sec31A在HNSCC组织中的表达情况,利用Kaplan-Meier生存分析比较Sec31A与HNSCC患者预后的关系;在HNSCC细胞系HN6、HN4 中转染小干扰质粒 si-Sec31A、si-NC,通过 CCK-8 实验、平板克隆实验、划痕愈合实验和 Transwell 实验检测Sec31A敲低后对HNSCC细胞生物学行为的影响;通过蛋白免疫印迹(Western Blot,WB)实验检测HN6 及HN4 敲低Sec31A后细胞中PI3K/AKT/mTOR通路相关蛋白表达水平的变化;构建HN6-siSec31A和HN6-siNC的转染稳定细胞株接种于裸鼠皮下进一步验证Sec31A在体内的成瘤作用.结果 TCGA数据显示Sec31A在HNSCC组织中高于癌旁正常组织(P<0.01),且Sec31A高表达与患者的预后不良显著相关(P<0.05),免疫组化染色显示Sec31A在HNSCC中表达强于正常组织;在HN6 和HN4 细胞中,敲低Sec31A后,细胞的增殖、迁移和侵袭能力明显弱于对照组;通过WB实验发现,HN6、HN4 转染si-Sec31A后,p-PI3K、p-AKT、p-mTOR蛋白表达水平显著降低;HN6 敲低Sec31A后在裸鼠皮下的移植瘤体积明显小于对照组.结论 敲低Sec31A后可以抑制HNSCC细胞增殖、迁移和侵袭的能力,可能是通过PI3K/AKT/mTOR通路发挥作用.
The effect of knocking down Sec31A on the malignant phenotype of HNSCC
Objective To explore the impact of knocking down Sec31A on the malignant phenotype of head and neck squamous cell carcinoma(HNSCC)and its possible mechanisms.Methods Transcriptome sequencing data of HNSCC tissues and adjacent tissues were obtained from the TCGA database,and the expression levels of Sec31A were compared.Immunohistochemical staining was used to analyze the expression of Sec31A in HNSCC tissues.Kaplan-Meier survival analysis was used to compare the relationship between Sec31A and the prognosis of HNSCC patients.Small interfering plasmids si-Sec31A and si-NC were transfected into HNSCC cell lines HN6 and HN4,and the impact of knocking down Sec31A on the biological behavior of HNSCC cells was detected through CCK-8 exper-iments,plate cloning experiments,scratch healing experiments,and Transwell experiments.Changes in the expression levels of PI3K/AKT/mTOR pathway related proteins in cells were detected after knocking down Sec31A with HN6 and HN4 through Western Blot(WB)experiments.Stable transfected cell lines of HN6 siSec31A and HN6 siNC were constructed and inoculated subcutaneously in nude mice to further verify the tumorigenic effect of Sec31A in vivo.Results TCGA data showed that Sec31A was higher in HNSCC tissues than in adjacent normal tissues(P<0.01),and high expression of Sec31A was significantly correlated with poor prognosis in pa-tients(P<0.05).Immunohistochemical staining showed that Sec31A was expressed stronger in HNSCC tissues than in normal tissues.In HN6 and HN4 cells,knocking down Sec31A resulted in significantly weaker proliferation,migration,and invasion abilities compared to the control group.Through WB experiments,it was found that transfection of si-Sec31A with HN6 and HN4 significantly reduced the expression levels of p-PI3K,p-AKT,and p-mTOR proteins.After knocking down Sec31A with HN6,the transplanted tumor volume in nude mice was significantly smaller than that in the control group.Conclusion Knocking down Sec31A can inhibit the proliferation,migration and invasion of HNSCC cells,possibly through the PI3K/AKT/mTOR pathway.

Sec31Ahead and neck squamous cell carcinomamolecular targeted therapyPI3K/AKT/mTOR pathway

何垚、赵振远、高腾、林鹏、陈以任、宋晓萌

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南京医科大学口腔疾病重点实验室,江苏南京(210029)

南京医科大学附属口腔医院口腔颌面外科,江苏南京(210029)

Sec31A 头颈鳞状细胞癌 分子靶向治疗 PI3K/AKT/mTOR通路

国家自然科学基金

81772887

2024

口腔医学
南京医科大学口腔医学院

口腔医学

CSTPCD
影响因子:0.641
ISSN:1003-9872
年,卷(期):2024.44(7)
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