摘要
目的:探讨SMC4在人食管癌(EC)组织中的表达及其对EC细胞增殖、侵袭和转移的作用机制。方法:收集2018年5月至2019年4月我院手术切除的EC组织标本及其癌旁正常组织标本各114例,免疫组化法检测人EC组织和正常食管组织中SMC4表达,并分析其与食管癌临床病理学参数的关系,采用细胞转染技术构建稳定下调SMC4的KYSE170和Eca-109细胞(KYSE170-siSMC4组/Eca-109-siSMC4组),以未经处理的KYSE170和Eca-109细胞(Control组)转染siNC的KYSE170/Eca-109细胞(KYSE170-siNC组/Eca-109-siNC组)作为对照,MTT法检测KYSE170和Eca-109细胞增殖;Transwell小室法检测转染后KYSE170和Eca-109细胞侵袭;划痕实验检测转染后KYSE170和Eca-109细胞迁移;IFA检测SMC4对上皮间质转化(EMT)相关分子N-cadherin表达的影响,Western blot检测细胞中PI3K、p-PI3K、Akt、p-Akt蛋白表达。结果:SMC4在食管癌组织中的阳性表达率高于正常食管黏膜组织(P<0.001);SMC4与肿瘤直径、TNM临床分期、浸润程度、淋巴结转移及组织分化程度密切相关(P<0.05);与Control组和KYSE170-siNC组相比,KYSE170-siSMC4组细胞在第4、5和6天增殖能力明显下降(P<0.05),Eca-109-siSMC4组4 d、5 d和6 d增殖能力明显低于Control组和Eca-109-siNC组(P<0.05);KYSE170-siSMC4组穿膜细胞数显著少于Control组和KYSE170-siNC组(P<0.05),Eca-109-siSMC4组穿膜细胞数明显少于Control组和Eca-109-siNC组(P<0.05);KYSE170-siSMC4组细胞迁移率明显低于Control组和KYSE170-siNC组(P<0.05),Eca-109-siSMC4组细胞迁移率明显低于Control组和Eca-109-siNC组(P<0.05);共聚焦显微镜观察发现,下调SMC4后,与Control组和KYSE170-siNC相比,KYSE170-siSMC4细胞N-cadherin表达明显下降(P<0.05);KYSE170-siSMC4组p-PI3K和p-Akt表达明显低于Control组和KYSE170-siNC组(P<0.05)。结论:下调SMC4表达可抑制人EC细胞增殖、侵袭和转移,并通过降低EMT相关分子N-cadherin表达抑制EMT,其作用机制可能与PI3K/AKT信号通路有关。
Objective:To investigate expression of SMC4 in human esophageal carcinoma(EC)tissue and its mechanism of proliferation,invasion and metastasis on EC cells.Methods:114 esophageal cancer tissue specimens and 114 corresponding adjacent normal tissue specimens surgically removed in our hospital from May 2018 to April 2019 were collected. Expressions of SMC4 in human EC tissue and normal tissue were detected by immunohistochemical method,and relationship between SMC4 expression and clinicopathological parameters of EC was analyzed. KYSE170 and Eca-109 cells(KYSE170-siSMC4 group/Eca-109-siSMC4 group)stably down-regulated SMC4 were constructed by cell transfection technology. Untreated KYSE170 and Eca-109 cells(Control group)and transfected siNC KYSE170/Eca-109 cells(KYSE170-siNC group/Eca-109-siNC group)were used as controls,and proliferation of KYSE170 and Eca-109 cells was detected by MTT method. Transwell chamber method to detect invasive of transfected KYSE170 and Eca-109 cells. Scratch test to detect migration of transfected KYSE170 and Eca-109 cells. IFA to detect effect of SMC4 on expression of N-cadherin,a molecule related to epithelial-mesenchymal transition(EMT). Western blot to detect expressions of PI3K,p-PI3K,Akt and p-Akt proteins in cells.Results:Positive expression rate of SMC4 in esophageal carcinoma tissue was higher than that in normal EC tissue(P<0.001). SMC4 was closely related to tumor diameter,TNM clinical stage,infiltration degree,lymph node metastasis and tissue differentiation degree(P<0.05). Compared with Control group and KYSE170-siNC group,proliferation ability of KYSE170-siSMC4 group cells was significantly decreased at 4,5 and 6 d(P<0.05). Proliferation ability of Eca-109-siSMC4 group at4,5 and 6 d was significantly lower than that of Control group and Eca-109-siNC group(P<0.05). Number of transmembrane cells in KYSE170-siSMC4 group was significantly lower than that in Control group and KYSE170-siNC group(P<0.05). Number of transmembrane cells in Eca-109-siSMC4 group was significantly low
作者
张冉
石长林
苟小军
何鸿晏
ZHANG Ran;SHI Chang-Lin;GOU Xiao-Jun;HE Hong-Yan(Department of Cardiothoracic Surgery,Bazhong Central Hospital of Sichuan Province,Bazhong 636000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2021年第11期1339-1345,共7页
Chinese Journal of Immunology
关键词
染色体结构维持蛋白4
人食管癌细胞
增殖
侵袭
转移
Structural maintenance of chromosome 4
Human esophageal cancer cells
Proliferation
Invasion
Metastatic
