PDRG1 基因沉默有助于抑制胃癌细胞的生长并诱导其凋亡。

PDRG1 gene silencing contributes to inhibit the growth and induce apoptosis of gastric cancer cells.

机构信息

Department of General Surgery, Beijing Anzhen Hospital, Capital Medical University, Beijing, China.

出版信息

Pathol Res Pract. 2019 Oct;215(10):152567. doi: 10.1016/j.prp.2019.152567. Epub 2019 Jul 27.

DOI:10.1016/j.prp.2019.152567

PMID:31383535

Abstract

OBJECTIVE

This paper aims to provide some experimental basis for unveiling the role of PDRG1 (P53 And DNA Damage-Regulated Gene 1) gene silencing in the growth and development of gastric cancer.

METHODS

PDRG1 levels in gastric cancer tissues and cell lines were measured by Western blotting. Then, gastric cancer BGC-823 cells, divided into Control, PDRG1 siRNA, NC siRNA and PDRG1 siRNA + KU55933 (ATM inhibitor) groups, were used to conduct a series of in vitro experiments including MTT, Flow cytometry, Wound-healing and Transwell assays. Expression of PDRG1 and ATM/p53 pathway-related proteins were determined by Western blot. Eventually, experiment in vivo was carried out to verify the control of PDRG1 on gastric cancer cells after establishing the tumor xenograft model in nude mice.

RESULTS

PDRG1 was significantly elevated in gastric cancer tissues and was associated with lower cell differentiation degree, more severe lymph node metastasis and higher tumor stage of gastric cancer patients. The growth of BGC-823 cells were significantly retarded and the cell apoptosis was increased in the PDRG1 siRNA group; besides, cell cycle was arrested in G2/M phase, and the expressions of p-ATM, p53, p21, p-cdc2 and cleaved caspase-3 were up-regulated with the reduced PDRG1. However, KU55933 could reverse the anti-tumor effect of PDRG1 siRNA on BGC-823 cells. The in-vivo experiment confirmed PDRG1 siRNA can inhibit tumor xenograft growth in nude mice.

CONCLUSION

Specific PDRG1 gene silencing may inhibit the growth and metastasis of gastric cancer cells through the activation of ATM/p53 pathway.

摘要

目的

本文旨在为揭示 PDRG1（P53 和 DNA 损伤调节基因 1）基因沉默在胃癌生长和发展中的作用提供一些实验依据。

方法

通过 Western blot 检测胃癌组织和细胞系中 PDRG1 的水平。然后，将胃癌 BGC-823 细胞分为对照组、PDRG1 siRNA 组、NC siRNA 组和 PDRG1 siRNA+KU55933（ATM 抑制剂）组，进行一系列体外实验，包括 MTT、流式细胞术、划痕愈合和 Transwell assays。Western blot 检测 PDRG1 和 ATM/p53 通路相关蛋白的表达。最后，建立裸鼠肿瘤异种移植模型，验证 PDRG1 对胃癌细胞的控制作用。

结果

PDRG1 在胃癌组织中明显升高，与胃癌患者细胞分化程度较低、淋巴结转移较严重、肿瘤分期较高有关。PDRG1 siRNA 组 BGC-823 细胞生长明显受阻，细胞凋亡增加；此外，细胞周期被阻滞在 G2/M 期，p-ATM、p53、p21、p-cdc2 和 cleaved caspase-3 的表达上调，PDRG1 减少。然而，KU55933 可以逆转 PDRG1 siRNA 对 BGC-823 细胞的抗肿瘤作用。体内实验证实 PDRG1 siRNA 可抑制裸鼠肿瘤异种移植的生长。