Wu Yunshi, Xue Jun, Li Yuanrui, Wu Xueliang, Qu Ming, Xu Dandan, Shi Yongquan
Graduate School of Hebei North University, Zhangjiakou, China.
Department of General Surgery, The First Affiliated Hospital of Hebei North University, Zhangjiakou, China.
J Gastrointest Oncol. 2021 Aug;12(4):1577-1589. doi: 10.21037/jgo-21-403.
The incidence of colorectal cancer is high and on the rise. The genetic and protein expressions of RUNT-associated transcription factor 3 (RUNX3) and human epidermal growth factor receptor 2 (HER2) in colorectal cancer (CRC) and adjacent normal tissues were detected to preliminarily explore their correlation and clinical significance.
CRC specimens excised during general surgery were selected for localization and quantitative analysis of protein and gene expression by SP (Streptavidin-peroxidase conjugated method) immunohistochemical staining, reverse transcription-polymerase chain reaction (RT-qPCR) and western blot. Combined with the patients' data, the relationship between the expression of the two genes and tumor characteristics was analyzed. Log-rank test was used to analyze the correlation between the two proteins and survival prognosis of CRC patients. The expression of RUNX3 in RKO and HCT-116 was knocked down, and the relative expression of HER2 in the two cell lines was detected.
Immunohistochemical, RT-qPCR and Western blot results showed that the positive expression rate of RUNX3 in CRC was lower than in the normal group, and HER2 in CRC was higher than in the normal group. The positive expression of the two proteins correlated with the pT and pN stages of CRC. A significant negative correlation between the expression of the two genes in CRC. Follow-up results showed that the number of -positive patients was higher than negative ones, while positive were fewer than negative ones. In vitro experiments showed that protein and gene expression decreased, HER2 protein and gene expression increased in knockdown RKO and HCT-116 cells, respectively (P<0.05).
The expression of and in CRC is related to the occurrence and development of CRC, and the two genes have a negative regulating effect.
结直肠癌发病率高且呈上升趋势。检测 runt 相关转录因子 3（RUNX3）和人表皮生长因子受体 2（HER2）在结直肠癌（CRC）及癌旁正常组织中的基因及蛋白表达，初步探讨其相关性及临床意义。
选取普通外科手术切除的 CRC 标本，采用链霉亲和素 - 过氧化物酶结合法（SP）免疫组化染色、逆转录 - 聚合酶链反应（RT - qPCR）及蛋白质印迹法进行蛋白和基因表达的定位及定量分析。结合患者资料，分析两个基因的表达与肿瘤特征的关系。采用对数秩检验分析两种蛋白与 CRC 患者生存预后的相关性。敲低 RKO 和 HCT - 116 细胞中 RUNX3 的表达，检测两种细胞系中 HER2 的相对表达。
免疫组化、RT - qPCR 和蛋白质印迹结果显示，CRC 中 RUNX3 的阳性表达率低于正常组，HER2 在 CRC 中的表达高于正常组。两种蛋白的阳性表达与 CRC 的 pT 和 pN 分期相关。CRC 中两个基因的表达呈显著负相关。随访结果显示，双阳性患者数量高于双阴性患者，而单阳性患者数量少于单阴性患者。体外实验表明，敲低 RKO 和 HCT - 116 细胞中的 RUNX3 后，HER2 蛋白和基因表达分别增加，RUNX3 蛋白和基因表达降低（P<0.05）。
RUNX3 和 HER2 在 CRC 中的表达与 CRC 的发生发展有关，且两个基因具有负调控作用。
