摘要
目的观察DNA损伤诱导转录物4(DDIT4)在大肠癌(CRC)组织和癌旁组织中的表达,探索其与临床病理特征的相关性、预后及作用机制。方法从癌症基因组图谱(TCGA)数据库收集CRC数据集,比较DDIT4mRNA的表达及预后相关性分析,筛选关键基因进行功能富集分析。收集2022年5月至2023年5月在温州医科大学黄岩医院诊断为CRC的石蜡标本观察组100例,对照组60例,通过免疫组化染色检测肿瘤和癌旁DDIT4的表达,并进行临床病理特征的相关性分析。培养CRC及正常肠上皮细胞系,RT-PCR检测其DDIT4 mRNA的表达,基因干预DDIT4的表达,Western blot法检测自噬通路相关蛋白(mTOR、p-mTOR、Beclin1、P62)的表达。结果TCGA数据库数据和免疫组化结果显示DDIT4在CRC中表达显著增加(P<0.001),与患者总体生存率、浸润深度、肿瘤大小、淋巴结转移、DUKES分期和血清CEA水平相关(P<0.05)。沉默DDIT4基因,mTOR自噬相关通路下游p-mTOR/mTOR比值及P62相对升高,Beclin1相对降低(P<0.05)。结论DDIT4基因与CRC的发生发展及不良预后显著相关,其机制可能与DDIT4参与调控mTOR信号通路激活自噬有关。
Objective To observe the expression of DNA damage-inducible transcript 4(DDIT4)in colorectal cancer(CRC)tissues and paracancerous tissues,and to explore the correlation with clinicopathologic features,prognosis and mechanism of action.Methods A complete CRC dataset was collected from The Cancer Genome Atlas(TCGA)database to compare the expression and prognostic correlation analyses of DDIT4 mRNA,and key genes were screened for functional enrichment analysis.100 cases in the observation group and 60 cases in the control group of paraffin specimens diagnosed with CRC in our hospital from May 2022 to May 2023 were collected,and the expression of tumor and paraneoplastic DDIT4 was detected by immunohistochemical staining and correlation analysis of clinicopathological features was performed.The human colon epithelial cell line was used as the control,and the CRC cell line was cultured,and the expression of DDIT4 mRNA was detected by RT-PCR,the expression of DDIT4 by gene intervention,and the expression of autophagy pathway-related proteins(mTOR,p-mTOR,Beclin1,P62)by Western blot.Results TCGA database data and immunohistochemistry results showed that DDIT4 expression was significantly increased in CRC(P<0.001),which was correlated with patients'overall survival,depth of infiltration,tumor size,lymph node metastasis,DUKES staging and serum CEA level(P<0.05).Silencing the DDIT4 gene resulted in a relative increase in p-mTOR/mTOR ratio and P62 and a relative decrease in Beclin1 downstream of the mTOR autophagy-related pathway(P<0.05).Conclusion DDIT4 gene was significantly associated with the development and poor prognosis of CRC,and its mechanism may be related to the involvement of DDIT4 in regulating the mTOR signaling pathway to activate autophagy.
作者
黄合
李剑敏
谭玲
游淑清
HUANG He;LI Jianmin;TAN Ling;YOU Shuqing(Wenzhou Medical University Huangyan Hospital,Zhejiang 318020,China)
出处
《浙江创伤外科》
2024年第6期1003-1008,共6页
Zhejiang Journal of Traumatic Surgery
关键词
DNA损伤诱导转录物4
自噬
大肠癌
免疫组化
不良预后
DNA damage-induced transcript 4
autophagy
colorectal cancer
immunohistochemistry
poor prognosis
