敲减辅酶Q10B表达对裸鼠食管鳞癌皮下移植瘤细胞增殖、凋亡及上皮间质转化的影响  

Effects of knocking down coenzyme Q10B on proliferation,apoptosis and EMT of esophageal squamous cell carcinoma xenografts in nude mice

作　　者：魏瑜[1] 李志芳 曹雷雨[1] 高艳 马晓丽[1] 张莉[1] WEI Yu;LI Zhifang;CAO Leiyu;GAO Yan;MA Xiaoli;ZHANG Li(The Fourth Department of General Internal Medicine/Special Needs Medicine,The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学第一附属医院综合内四科/特需内科,乌鲁木齐830054

出　　处：《山东医药》2024年第28期40-43,共4页Shandong Medical Journal

基　　金：省部共建中亚高发病成因与防治国家重点实验室开放课题资助项目(SKL-HIDCA-2022-SG1);新疆维吾尔自治区重点研发计划项目(2020B03003,2020B03003-3);新疆维吾尔自治区自然科学基金资助项目(2022D01C783)。

摘　　要：目的探讨敲减辅酶Q10B(COQ10B)表达对裸鼠食管鳞状细胞癌(鳞癌)皮下移植瘤细胞增殖、凋亡及上皮间质转化(EMT)的影响。方法利用慢病毒转染技术将COQ10B的干扰慢病毒及其阴性对照转染至食管鳞癌细胞(KYSE150细胞)中,构建敲减COQ10B表达的KYSE150细胞(sh-COQ10B组)和阴性对照KYSE150细胞(sh-NC组)。选择雌性BALB/c裸鼠20只,随机分入sh-COQ10B组和sh-NC组各10只,分别于裸鼠右前肩胛处皮下接种转染sh-COQ10B和sh-NC的KYSE150细胞,以此构建裸鼠皮下移植瘤模型,观察接种后不同时间皮下移植瘤生长情况并测量瘤体的体积和质量。接种第26天实验结束后取瘤体组织,采用HE、免疫组化、TUNEL染色法观察裸鼠皮下移植瘤中细胞增殖和凋亡的情况,以Westernblotting法检测移植瘤中EMT相关蛋白(E-cadherin和Vimentin)和凋亡相关蛋白(Bcl-2和Bax)表达。结果与sh-NC组比较,sh-COQ10B组中COQ10B蛋白表达量低(P<0.05);皮下移植瘤模型构建成功且裸鼠均无死亡。sh-COQ10B组移植后不同时间瘤体体积和质量均小于sh-NC组(P均<0.05)。与sh-NC组比较,sh-COQ10B组肿瘤细胞凋亡率高,瘤体组织内Bcl-2、Vimentin蛋白表达低,Bax、E-cadherin表达高,差异均有统计学意义(P均<0.05)。结论敲减COQ10B表达可抑制裸鼠食管鳞癌皮下移植瘤细胞增殖和EMT的发生,促进肿瘤细胞凋亡。Objective To investigate the effects of coenzyme Q10B(COQ10B)knockdown on the proliferation,apoptosis and epithelial-mesenchymal transition(EMT)of esophageal squamous cell carcinoma xenografts in nude mice.Methods The interference lentivirus of COQ10B and its negative control were transfected into KYSE150 cells by lentivi-rus transfection technology.The KYSE150 cells with knockdown of COQ10B expression(sh-COQ10B group)and the nega-tive control KYSE150 cells(sh-NC group)were constructed.Twenty female BALB/c nude mice were randomly divided in-to the sh-COQ10B group(n=10)and sh-NC group(n=10).KYSE150 cells transfected with sh-COQ10B and sh-NC were subcutaneously injected into the right anterior scapula of nude mice to establish subcutaneous transplantation tumor mod-els.The growth of subcutaneous xenografts at different time after inoculation was observed and the size and the weight of tu-mors were measured.At the end of the experiment on the 26th day after inoculation,tumor tissues were taken,HE,immu-nohistochemistry,and TUNEL staining methods were used to observe cell proliferation and apoptosis in subcutaneous xeno-grafts in nude mice.Western blotting was used to detect the expression levels of EMT-related proteins(E-cadherin and Vi-mentin)and apoptosis-related proteins(Bcl-2 and Bax).Results Compared with the sh-NC group,the protein expres-sion of COQ10B in the sh-COQ10B group decreased(P<0.05).The subcutaneous transplanted tumor models were estab-lished successfully and no death occurred in nude mice.The tumor volume and mass in the sh-COQ10B group were lower than those in the sh-NC group(both P<0.05).Compared with the sh-NC group,the apoptosis rate of tumor cells in the sh-COQ10B group increased,the protein expression levels of Bcl-2 and Vimentin in tumor tissues decreased,and the expres-sion levels of Bax and E-cadherin increased,with statistically significant difference(all P<0.05).Conclusion Knock-ing down COQ10B expression can inhibit the proliferation of subcutaneous transplanted tumor and the occurrence

关 键 词：食管鳞状细胞癌 裸鼠皮下移植瘤 辅酶Q10B 细胞增殖 细胞凋亡 上皮间质转化 

分 类 号：R735.1[医药卫生—肿瘤]