摘要
目的探讨鸟嘌呤核苷酸结合蛋白α亚基12(G protein subunitα12,GNA12)对鼻咽癌细胞生物学行为的影响。方法利用GEPIA2、Human Protein Altas数据库预测GNA12在头颈部肿瘤中的表达情况,利用Keplan-Meier plotter数据库分析GNA12表达与头颈部肿瘤预后的关系。采用qRT-PCR实验检测正常鼻咽黏膜上皮细胞系(NP69)和鼻咽癌细胞系(CNE-2、5-8F、HK-1)中GNA12的表达水平。用携带si-GNA12和NC的干扰序列分别转染鼻咽癌细胞CNE-2和5-8F细胞系,构建敲低的细胞株(si-GNA12组)和对照细胞株(NC组)。采用划痕实验和Transwell实验检测细胞迁移能力,CCK-8和集落克隆实验检测细胞的增殖能力。采用Western blot实验检测上皮-间质转化中N-cadherin、E-cadherin、Snail以及Vimentin蛋白和ERK信号通路中ERK及p-ERK蛋白表达情况。结果数据库结果显示,头颈部肿瘤组织中GNA12表达高于正常头颈部组织(P<0.05)。GNA12表达越高,预后越差(P<0.05)。与正常鼻咽黏膜上皮细胞相比,鼻咽癌细胞系中GNA12 mRNA表达水平较高(P<0.05)。与NC组相比,si-GNA12组GNA12 mRNA表达水平下降(P<0.05)。与NC组相比,si-GNA12组CNE-2和5-8F细胞的增殖、迁移和侵袭均受到抑制(P<0.05)。Western blot显示,与NC组相比,si-GNA12组N-cadherin、Vimentin、Snail蛋白表达下降(P<0.05),E-cadherin蛋白表达上升(P<0.05),ERK信号通路相关蛋白p-ERK蛋白表达下降(P<0.05)。结论GNA12可能通过调控ERK信号通路促进鼻咽癌细胞的增殖和侵袭迁移。
Objective To investigate the effect of G protein subunitα12(GNA12)on the biological behaviors of nasopharyngeal carcinoma cells.Methods The expression of GNA12 in head and neck tumors was predicted using GEPIA2 and Human Protein Altas databases,and the relationship between GNA12 expression and the prognosis of head and neck tumors was analyzed by Keplan-Meier plotter database.The expression levels of GNA12 in normal nasopharyngeal mucosal epithelial cell lines(NP69)and nasopharyngeal carcinoma cell lines(CNE-2,5-8F,HK-1)were detected by qRT-PCR assay.Nasopharyngeal carcinoma cells CNE-2 and 5-8F were transfected with the interfering sequences carrying si-GNA12 and NC to construct knockdown cell lines(si-GNA12 group)and control cell lines(NC group),respectively.Cell migration ability was detected using wound healing assay and Transwell assay,and cell proli-feration ability was detected using CCK-8 and colony cloning assay.Western blot assay was used to detect the expressions of N-cadherin,E-cadherin,Snail,and Vimentin proteins in the epithelial-mesenchymal transition,and ERK and p-ERK proteins in the ERK signaling pathway.Results According to the databases,GNA12 expression in head and neck tumors was higher than that in normal head and neck tissues(P<0.05).The higher the GNA12 expression was,the worse the prognosis was(P<0.05).Compared with normal nasopharyngeal mucosal epithelial cells,GNA12 mRNA levels were increased in nasopharyngeal carcinoma cell lines(P<0.05).Compared with NC group,the expression level of GNA12 mRNA was decreased in si-GNA12 group(P<0.05),and cell proliferation,migration,and invasion were inhibited(P<0.05).Western blot result showed that compared with NC group,the protein expressions of N-cadherin,Vimentin and Snail were decreased in si-GNA12 group(P<0.05),the protein expression of E-cadherin was increased(P<0.05),and the expression of p-ERK protein was decreased(P<0.05).Conclusion GNA12 can promote the proliferation,invasion and migration of nasopharyngeal carcinoma cells by regulating the
作者
张坤
李敏
葛思辰
蒋成义
ZHANG Kun;LI Min;GE Sichen;JIANG Chengyi(Department of Otorhinolaryngology,Head and Neck Surgery,First Affiliated Hospital of Bengbu Medical University,Bengbu 233000,China)
出处
《山西医科大学学报》
CAS
2024年第3期293-302,共10页
Journal of Shanxi Medical University
基金
安徽省重点研究与开发项目:人口健康专项(202104j07020015)
蚌埠医学院研究生科研创新计划项目(Byyczx22052)。
