摘要
目的探究分泌型卷曲相关蛋白4(SFRP4)在DNA错配修复(MMR)功能正常与MMR功能缺陷的Ⅱ期结肠癌中的表达及意义。方法收集新鲜的Ⅱ期结肠癌组织标本,并根据MMR状态分为pMMR和dMMR组。利用Affymetrix Human oeLncRNA基因芯片检测两组不同MMR状态结肠癌组织中差异表达的mRNA情况。通过q-PCR和Westernblot技术检测dMMR和pMMR肠癌组织、细胞株中SFRP4的表达并分析两组间表达差异情况;通过免疫组化技术检测增殖标志物Ki-67的表达并分析SFRP4与Ki-67表达的相关关系。利用HCT116细胞株,干扰SFRP4的表达,通过流式细胞术检测干扰SFRP4前与干扰后HCT116细胞株的凋亡率并分析其差异。结果 PCR和Western blot的结果提示SFRP4在dMMR状态的肠癌组织、细胞株中的表达明显高于pMMR的肠癌组织(P=0.014)、细胞株(P=0.0079),免疫组化结果提示SFRP4与Ki-67蛋白在肠癌组织的表达呈负相关关系(P=0.041)。siRNA干扰SFRP4的表达后HCT116细胞株凋亡率下降,包括早期凋亡(P=0.003)和晚期凋亡率(P=0.024)。结论 SFRP4在dMMR型肠癌表达较pMMR型表达增高,促进肠癌细胞凋亡、抑制增殖,可能有助于改善MMR缺陷型Ⅱ期肠癌预后。
Objective To investigate the expressions of secreted frizzled-related protein 4(SFRP4) in stage II DNA mismatch repair-deficient(dMMR) and mismatch repair-proficient(pMMR) colorectal cancers and explore their clinical significance.Methods We collected fresh stage II colon cancer tissues with different MMR status detected by immunohistochemistry(IHC).The differentially expressed mRNAs between dMMR and pMMR tumors were identified by Affymetrix Human oeLncRNA gene chip, and the expression of SFRP4 in these cancer tissues and in colorectal cancer cell lines were detected using Western blotting and real-time quantitative PCR. The apoptosis rates of HCT116 cells with and without siRNA-mediated transient SFRP4 knockdown were determined using flow cytometry. We further investigated the expression pattern of Ki-67 and its correlation with SFRP4 expression. Results Compared with pMMR colon cancer tissues or cells, both dMMR colon cancer tissues(P=0.014) and cells(P=0.0079) showed significantly increased expression of SFRP4, which was in negative correlation with Ki-67(P=0.041). In HCT116 cells, transient SFRP4 knockdown resulted in decreased cell apoptosis, including both early apoptosis(P=0.003) and late apoptosis(P=0.024). Conclusion Up-regulation of SFRP4 in dMMR stage II colon cancer promotes apoptosis and inhibits proliferation of the cancer cells, and may improve the prognosis of dMMR colon cancer.
作者
陈可绪
梁汉霖
彭杰文
郑燕芳
CHEN Kexu;LIANG Hanlin;PENG Jiewen;ZHENG Yanfang(Center of Oncology,Zhongshan People's Hospital,Zhongshan 528400,China;Center of Oncology,Zhujiang Hospital,Southern Medical University,Guangzhou 510282,China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2018年第11期1300-1305,共6页
Journal of Southern Medical University
基金
中山市科技计划项目(2017B1037)
博士后科研启动基金(2015B0690)
关键词
肠癌
错配修复
分泌型卷曲相关蛋白4
增殖
凋亡
colorectal cancer
mismatch repair
secreted frizzled-related protein 4
proliferation
apoptosis
