Department of Colorectal Surgery, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Institute, NO.44 Xiaoheyan Road, Dadong District, Shenyang, 110042, People's Republic of China.
Biol Res. 2018 Nov 24;51(1):51. doi: 10.1186/s40659-018-0200-9.
Emerging evidence showed that microRNAs (miRs) play critical roles in human cancers by functioning as either tumor suppressor or oncogene. MIR-382 was found to function as tumor suppressor in certain cancers. However, the role of MIR-382 in colorectal cancer (CRC) is largely unknown. Specificity protein 1 (SP1) is highly expressed in several cancers including CRC and is correlated with poor prognosis, but it is unclear whether or not MIR-382 can regulate the expression of SP1.
MIR-382 expression level was measured by reverse transcription-quantitative polymerase chain reaction. The connection between MIR-382 and SP1 was validated by luciferase activity reporter assay and western blot assay. Cell counting kit-8 assay and wound-healing assay were conducted to investigate the biological functions of MIR-382 in CRC.
In this study, we found MIR-382 expression was downregulated in CRC tissues and cell lines, and the transfection of MIR-382 mimic decreased cell growth and migration. Furthermore, we identified SP1 was a direct target of MIR-382. Overexpression of MIR-382 decreased the expression of SP1, whereas MIR-382 knockdown promoted SP1 expression. We also observed an inversely correlation between MIR-382 and SP1 in CRC tissues. Additionally, we showed that knockdown of SP1 inhibited cell growth and migration and attenuated the effect of MIR-382 inhibitor on cell behaviors.
In conclusion, the present study describes a potential mechanism underlying a MIR-382/SP1 link contributing to CRC development. Thus, MIR-382 may be able to be developed as a novel treatment target for CRC.
新出现的证据表明,微小 RNA(miRs)通过作为肿瘤抑制因子或癌基因发挥关键作用,在人类癌症中发挥作用。已经发现 MIR-382 在某些癌症中作为肿瘤抑制因子发挥作用。然而,MIR-382 在结直肠癌(CRC)中的作用在很大程度上是未知的。特异性蛋白 1(SP1)在包括 CRC 在内的几种癌症中高度表达,并且与预后不良相关,但尚不清楚 MIR-382 是否可以调节 SP1 的表达。
通过逆转录定量聚合酶链反应测量 MIR-382 的表达水平。通过荧光素酶活性报告基因测定和 Western blot 测定验证 MIR-382 与 SP1 之间的联系。通过细胞计数试剂盒-8 测定和划痕愈合测定研究 MIR-382 在 CRC 中的生物学功能。
在这项研究中,我们发现 MIR-382 在 CRC 组织和细胞系中表达下调,并且 MIR-382 模拟物的转染降低了细胞生长和迁移。此外,我们确定 SP1 是 MIR-382 的直接靶标。过表达 MIR-382 降低了 SP1 的表达,而 MIR-382 敲低则促进了 SP1 的表达。我们还观察到 CRC 组织中 MIR-382 和 SP1 之间存在负相关。此外,我们表明 SP1 的敲低抑制了细胞生长和迁移,并减弱了 MIR-382 抑制剂对细胞行为的影响。
总之,本研究描述了 MIR-382/SP1 联系在 CRC 发展中的潜在机制。因此,MIR-382 可能能够作为 CRC 的新型治疗靶点。