摘要
目的探讨SPANX家族成员C(SPANXC)过表达对头颈部鳞状细胞癌(HNSCC)Tu686细胞增殖与侵袭的影响。方法将Tu686细胞分为过表达(OE)-SPANXC组、NC组、Con组,对各组Tu686细胞进行转染,其中OE-SPANXC组加入GV358-SPANXC质粒,NC组加入空载体GV358,Con组只加等体积的培养基,共转染48 h。转染后收集细胞,采用实时荧光定量PCR检测Tu686细胞的SPANXC mRNA相对表达量;采用Western blotting检测SPANXC、细胞瘤病毒致癌基因同源物(c-Myc)、G1/S-特异性周期蛋白-D1(Cyclin D1)、β-连环蛋白(β-catenin)蛋白相对表达量;采用细胞计数试剂盒-8(CCK-8)细胞增殖实验和Transwell侵袭实验检测Tu686细胞的增殖和侵袭能力。结果OE-SPANXC组Tu686细胞SPANXC mRNA和蛋白的相对表达量高于Con组和NC组(P<0.05),提示SPANXC过表达的Tu686细胞构建成功。OE-SPANXC组细胞侵袭数量多于Con组和NC组(P<0.05)。培养5 d时CCK-8检测,OE-SPANXC组的细胞光密度值高于Con组和NC组(P<0.05)。OE-SPANXC组c-Myc、Cyclin D1和β-catenin蛋白相对表达量高于Con组和NC组(P<0.05)。结论SPANXC过表达可能通过上调c-Myc、Cyclin D1和β-catenin等促癌蛋白的表达,增强Tu686细胞的侵袭和增殖能力。
Objective To investigate the effect of overexpression of SPANX family member C(SPANXC)on the proliferation and invasion of Tu686 cells in head and neck squamous cell carcinoma(HNSCC).Methods Tu686 cells were divided into the overepression(OE)-SPANXC group,NC group and Con group,and transfected with corresponding reagents,respectively.Specifically,the OE-SPANXC group was transfected with GV358-SPANXC plasmid,the NC group with empty GV358 vector,and the Con group with an equal volume of culture medium only,with the total transfection duration of 48 h.Cells were collected after transfection.The relative expression level of SPANXC mRNA in Tu686 cells was detected by quantitative real-time PCR.Western blotting was performed to determine the relative protein expression levels of SPANXC,cellular myelocytomatosis oncogene homolog(c-Myc),G1/S-specific cyclin D1(Cyclin D1)andβ-catenin.The proliferation and invasion abilities of Tu686 cells were assessed via the Cell Counting Kit-8(CCK-8)cell proliferation assay and Transwell invasion assay,respectively.Results The relative expression levels of SPANXC mRNA and protein in Tu686 cells of the OE-SPANXC group were significantly higher than those in the Con and NC groups(P<0.05),indicating that the Tu686 cell line with SPANXC overexpression was successfully established.The number of invasive cells in the OE-SPANXC group was significantly greater than that in the Con and NC groups(P<0.05).At 5 days of culture,the optical density values of cells in the OE-SPANXC group were significantly higher than those in the Con and NC groups as detected by the CCK-8 assay(P<0.05).The relative protein expression levels of c-Myc,Cyclin D1 andβ-catenin in the OE-SPANXC group were significantly higher than those in the Con and NC groups(P<0.05).Conclusion SPANXC overexpression may enhance the proliferation and invasion abilities of Tu686 cells by upregulating the expression of oncogenic proteins including c-Myc,Cyclin D1 andβ-catenin.,
作者
肖荩茹
于海鸿
李华玲
陆合明
XIAO Jinru;YU Haihong;LI Hualing;LU Heming(Graduate School,Guangxi University of Chinese Medicine,Nanning 530000,Guangxi,China;Graduate School,Guilin Medical University,Guilin 541000,Guangxi,China;Clinical Cancer Center,the People's Hospital of Guangxi Zhuang Autonomous Region,Nanning 530000,Guangxi,China)
出处
《微创医学》
2025年第6期645-650,共6页
Journal of Minimally Invasive Medicine
基金
北京市希思科临床肿瘤学研究基金会课题(编号:Y-NESTLE2022MS-0180)。
关键词
头颈部鳞状细胞癌
SPANX家族成员C
细胞增殖
细胞侵袭
促癌蛋白
细胞实验
Head and neck squamous cell carcinoma
SPANXC family member C
Cell proliferation
Cell invasion
Oncogenic protein
Cell experiment
