南方医科大学学报

南方医科大学学报 ›› 2018, Vol. 38 ›› Issue (11): 1300-.doi: 10.12122/j.issn.1673-4254.2018.11.04

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分泌型卷曲相关蛋白4在不同DNA错配修复功能状态Ⅱ期肠癌中的表达及意义

陈可绪，梁汉霖，彭杰文，郑燕芳   

  • 出版日期:2018-11-20 发布日期:2018-11-20

Expression of secreted frizzled-related protein 4 in DNA mismatch repair-deficient and mismatch repair-proficient colorectal cancers

  • Online:2018-11-20 Published:2018-11-20

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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。

Abstract: 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.