miR-141/Sirt1 在结肠癌中的作用。

The role of miR-141/ Sirt1 in colon cancer.

机构信息

Second Department of Gastrointestinal Surgery, The Affiliated Hospital of Putian University, Putian, China.

出版信息

J BUON. 2020 Nov-Dec;25(6):2665-2671.

Abstract

PURPOSE

To explore the effects of micro ribonucleic acid-141 (miR)-141 on the proliferation and apoptosis of colon cancer cells and its association with the sirtuin 1 (Sirt1) expression.

METHODS

The samples of stage I, II, III and IV colon cancer were obtained, and the miRNA expression levels was analyzed, with normal colon tissues as controls. The expression of miR-141 and miR-34 was detected via quantitative reverse transcription-polymerase chain reaction (qRT-PCR), and the cell proliferation and apoptosis in each group were detected via cell counting kit-8 (CCK8) assay, respectively. Finally, the protein expressions of Sirt1, Caspase-3 and Caspase-8 were determined using Western blotting.

RESULTS

The expressions of miR-141 and miR-34 (miR-34 is mentioned in previous methods. Furthermore, we found the expression of miR-141 increasing with the progression of colon cancer, which was higher in stage III than in stage I-II and also higher in stage IV than in stage III. miR-34 was also highly expressed in stage IV colon cancer in our study were up-regulated in the progression of colon cancer. Overexpression of miR-141 could promote cell proliferation (p<0.05) and inhibit apoptosis (p<0.05), while inhibition on miR-141 expression could significantly weaken cell proliferation (p<0.05) and promote apoptosis (p<0.05). The results of luciferase reporter assay showed that miR-141 obviously inhibited Sirt1 (p<0.05). SRT2183 reduced cell proliferation (p<0.05) but up-regulated the protein expressions of Sirt1, Caspase-3 and Caspase-8 (p<0.05), while EX 527 had the opposite effects (p<0.05).

CONCLUSIONS

MiR-141 may promote proliferation and reduce apoptosis of colon cancer cells via targeting Sirt1.

摘要

目的

探讨微小 RNA-141（miR-141）对结肠癌细胞增殖和凋亡的影响及其与沉默信息调节因子 1（Sirt1）表达的关系。

方法

获取 I 期、II 期、III 期和 IV 期结肠癌样本，并分析 miRNA 表达水平，以正常结肠组织作为对照。通过定量逆转录聚合酶链反应（qRT-PCR）检测 miR-141 和 miR-34 的表达，分别通过细胞计数试剂盒-8（CCK8）检测各组细胞的增殖和凋亡。最后，用 Western blot 法测定 Sirt1、Caspase-3 和 Caspase-8 的蛋白表达。

结果

miR-141 和 miR-34 的表达（miR-34 在之前的方法中提到。此外，我们发现 miR-141 的表达随着结肠癌的进展而增加，III 期比 I-II 期高，IV 期比 III 期高。在我们的研究中，miR-34 在 IV 期结肠癌中也高表达。在结肠癌的进展过程中，miR-141 的过表达可促进细胞增殖（p<0.05）和抑制凋亡（p<0.05），而抑制 miR-141 的表达可显著减弱细胞增殖（p<0.05）和促进凋亡（p<0.05）。荧光素酶报告基因实验结果表明，miR-141 明显抑制 Sirt1（p<0.05）。SRT2183 减少细胞增殖（p<0.05），但上调 Sirt1、Caspase-3 和 Caspase-8 的蛋白表达（p<0.05），而 EX 527 则有相反的效果（p<0.05）。