整合素连接激酶在食管鳞状细胞癌组织中的表达及其对KYSE-150细胞增殖、凋亡及裸鼠移植瘤生长的影响  

Expression of integrin-lined kinase in esophageal squamous cell carcinoma tissues and its effect on proliferation and apoptosis of KYSE-150 cells and the growth of xenografts in nude mice

作　　者：马晓丽[1] 高艳 魏瑜[1] 曹雷雨[1] 张周华 张莉[1] MA Xiaoli;GAO Yan;WEI Yu;CAO Leiyu;ZHANG Zhouhua;ZHANG Li(The Fourth Department of General Internal Medicine,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,Xinjiang,China;The First Clinical Medical College,Xinjang Medical University,Urumqi 830054,Xinjang,China)

机构地区：[1]新疆医科大学第一附属医院综合内四科,新疆乌鲁木齐830054 [2]新疆医科大学第一临床医学院,新疆乌鲁木齐830054

出　　处：《中国肿瘤生物治疗杂志》2022年第6期549-556,共8页Chinese Journal of Cancer Biotherapy

基　　金：新疆维吾尔自治区自然科学基金资助项目(No.2020D01C258)。

摘　　要：目的:分析整合素连接激酶(ILK)基因在食管鳞状细胞癌(ESCC)组织中的表达水平及其与患者临床病理特征之间的关系,探讨其对KYSE-150细胞增殖、凋亡和裸鼠皮下移植瘤生长的影响。方法:选取2012年1月至2014年12月手术切除并经病理证实的75例ESCC患者的癌组织和其配对的癌旁组织标本,用组织芯片技术及免疫组织化学染色法检测ESCC组织和癌旁组织中ILK的表达情况;qPCR法检测ESCC细胞ECA109、TE-1、EC9706、KYSE-150中ILK mRNA的表达,选用ILK表达最高的KYSE-150细胞进行后续细胞功能学研究。使用ILK干扰慢病毒感染KYSE-150细胞下调ILK的表达,qPCR和WB法检测ILK基因敲降效率;MTT实验、克隆形成实验和FACS检测干扰ILK表达对KYSE-150细胞增殖能力和凋亡水平的影响;裸鼠皮下成瘤实验检测干扰ILK对KYSE-150细胞移植瘤生长的影响。结果:ESCC组织中ILK蛋白阳性表达率高于癌旁组织(P<0.05),且ILK高表达与淋巴结转移有关联(P<0.05)。ILK干扰慢病毒感染的KYSE-150细胞中ILK mRNA表达明显受到抑制(P<0.05),ILK蛋白水平表达下调,以上结果提示ILK敲降成功。与感染阴性对照病毒的KYSE-150细胞相比,ILK干扰慢病毒感染的KYSE-150细胞的增殖能力、克隆形成数均显著降低(均P<0.05),但细胞凋亡率升高(P<0.05)。与对照组相比,干预组裸鼠移植瘤生长缓慢,移植瘤的质量及体积均较小(均P<0.05)。结论:ESCC组织中ILK的表达高于癌旁组织,且ILK高表达与患者发生淋巴结转移有关联;抑制ILK基因可导致KYSE-150细胞增殖能力降低,促进细胞凋亡而抑制裸鼠移植瘤生长。Objective: To analyze the expression level of integrin-linked kinase(ILK) gene in esophageal squamous cell carcinoma(ESCC) tissues and its relationship with the clinicopathological characteristics of patients, and to explore its effect on KYSE-150 cell proliferation, apoptosis and the growth of subcutaneous xenograft tumor in nude mice. Methods: The cancer tissues and paired paracancerous tissues of 75 patients with ESCC who had surgical resection and confirmed by pathological examination from January 2012 to December 2014 were selected. Tissue chip technology and immunohistochemical staining were used to detect the expression of ILK in ESCC tissues and para-cancerous tissues;qPCR was used to detect the ILK expression in ESCC cell ECA109, TE-1, EC9706 and KYSE-150, and the KYSE-150 cells with the highest ILK expression was selected for subsequent cell functional studies. The KYSE-150 cells were transfected with ILK interference lentivirus to down-regulate the expression of ILK, and the knockdown efficiency was verified by qPCR and WB methods;the effects of interfering ILK expression on the proliferation and apoptosis of KYSE-150 cells were detected by MTT assay, clone formation assay and FACS;subcutaneous tumorigenesis assay in nude mice was used to detect the effect of interfering ILK on the growth of KYSE-150 cells transplanted tumors. Results: The positive rate of ILK protein in ESCC tissues was higher than that in para-cancerous tissues(P<0.05), and the high expression of ILK was associated with lymph node metastasis(P<0.05). The mRNA expression of ILK in shILK-infected KYSE-150 cells was significantly inhibited(P<0.05)and the expression of ILK protein was significantly down-regulated in the shILK group, which indicated the successful knockdown of ILK. Compared with the cells transfected with shCtrl, the proliferation ability and colony formation number of KYSE-150 cells in the shILK group were significantly reduced(both P<0.05), but the apoptosis rate was significantly increased(P<0.05). Compared with the NC

关 键 词：食管鳞状细胞癌 KYSE-150细胞 整合素连接激酶 SIRNA 增殖 凋亡 裸鼠 

分 类 号：R735.1[医药卫生—肿瘤] R730.2[医药卫生—临床医学]