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lncRNA HCP5的敲低通过miR-299-3p/PFN1/AKT轴抑制结直肠癌的进展。

Knockdown of lncRNA HCP5 Suppresses the Progression of Colorectal Cancer by miR-299-3p/PFN1/AKT Axis.

作者信息

Bai Ni, Ma Ying, Zhao Jia, Li Bo

机构信息

Department of Laboratory Medicine, The Affiliated Xi'an Centre Hospital of Xi'an Jiaotong University College of Medicine, Xi'an, Shaanxi 710003, People's Republic of China.

出版信息

Cancer Manag Res. 2020 Jun 19;12:4747-4758. doi: 10.2147/CMAR.S255866. eCollection 2020.

Abstract

BACKGROUND

Colorectal cancer (CRC) is one of the most common malignant tumors in the digestive system. The lncRNA HCP5 has been reported to affect the progression of tumor in several types of cancer. Here, in this research, we focus on the role and function of lncRNA HCP5 in human colorectal cancer.

MATERIALS AND METHODS

Tissue samples from colorectal cancer patients were used for detecting the expression of HCP5 by qRT-PCR. Proliferation, migration, invasion and apoptotic cells were assessed by CCK-8, colony formation, transwell assays and flow cytometry in SW480 and HCT-116 cells. The interactions between miR-299-3p and HCP5 or PFN1 were analyzed and confirmed by online database and luciferase reporter assays. The changes in PFN1 and AKT proteins were measured by Western blot. In vivo experiment was used to confirm the role of HCP5 in CRC.

RESULTS

The expression of HCP5 had a higher level in colorectal cancer samples and cells by qRT-PCR, comparing with the normal colorectal tissues and human normal colon epithelial cell. It was revealed that knockdown of HCP5 inhibited viabilities, migration and invasion, while inducing apoptosis in SW480 and HCT-116 cells. Then, HCP5 negatively regulated the expressions of miR-299-3p, which negatively regulated the expressions of PFN1 by targeting PFN1. Furthermore, miR-299-3p inhibitor could alleviate the inhibiting effect by si-HCP5 on cell process of SW480 and HCT-116 cells. In addition, the lncHCP5/miR-299-3p/PFN1 axis could affect the progression of CRC through activating the AKT signaling. Last, we confirmed that knockdown of HCP5 inhibited the progression of CRC with an in vivo experiment.

CONCLUSION

The experiments and analyses support our hypothesis that knockdown of lncRNA HCP5 suppresses the progression of colorectal cancer by miR-299-3p/PFN1/AKT axis.

摘要

背景

结直肠癌(CRC)是消化系统最常见的恶性肿瘤之一。据报道,lncRNA HCP5在几种癌症类型中影响肿瘤进展。在本研究中,我们聚焦于lncRNA HCP5在人类结直肠癌中的作用和功能。

材料与方法

采用来自结直肠癌患者的组织样本,通过qRT-PCR检测HCP5的表达。运用CCK-8、集落形成、Transwell实验和流式细胞术评估SW480和HCT-116细胞的增殖、迁移、侵袭和凋亡情况。通过在线数据库和荧光素酶报告基因实验分析并证实miR-299-3p与HCP5或PFN1之间的相互作用。采用蛋白质免疫印迹法检测PFN1和AKT蛋白的变化。通过体内实验证实HCP5在结直肠癌中的作用。

结果

与正常结直肠组织和人正常结肠上皮细胞相比,qRT-PCR检测显示HCP5在结直肠癌样本和细胞中的表达水平更高。研究发现,敲低HCP5可抑制SW480和HCT-116细胞的活力、迁移和侵袭,同时诱导细胞凋亡。随后,HCP5负向调节miR-299-3p的表达,而miR-299-3p通过靶向PFN1负向调节PFN1的表达。此外,miR-299-3p抑制剂可减轻si-HCP5对SW480和HCT-116细胞的细胞进程的抑制作用。另外,lncHCP5/miR-299-3p/PFN1轴可通过激活AKT信号通路影响结直肠癌的进展。最后,我们通过体内实验证实敲低HCP5可抑制结直肠癌的进展。

结论

实验和分析支持我们的假设,即敲低lncRNA HCP5通过miR-299-3p/PFN1/AKT轴抑制结直肠癌的进展。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/425c/7310975/9f491597c67e/CMAR-12-4747-g0001.jpg

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