piR-9994对胃癌细胞增殖、迁移及侵袭的影响及其机制

林华妹; 邹长棪; 苏颖; 胡丹; 廖锦容; 林可焴; 何火聪; 郑雄伟; 林贤东

doi:10.3971/j.issn.1000-8578.2021.21.0468

piR-9994对胃癌细胞增殖、迁移及侵袭的影响及其机制

Effect of piR-9994 on Proliferation, Migration and Invasion of Gastric Cancer Cells and Its Mechanism

摘要

摘要:
目的探讨piR-9994对胃癌细胞生物学行为的影响及可能机制。
方法 qRT-PCR检测piR-9994在胃癌细胞系（MGC803和AGS）和正常胃上皮细胞（GES-1）中的表达情况；构建过表达和敲低piR-9994的MGC803胃癌细胞株，MTT、克隆形成实验检测piR-9994表达变化对细胞增殖能力的影响，划痕愈合实验和Transwell侵袭实验检测细胞迁移和侵袭能力。qRT-PCR和Western blot检测细胞增殖和EMT相关基因表达。
结果 piR-9994在胃癌细胞MGC803中表达水平显著高于正常人胃黏膜上皮细胞GES-1（P < 0.05）。piR-9994过表达促进胃癌细胞增殖、侵袭和转移，敲低则作用相反。piR-9994表达与细胞周期密切相关，特别是S期。piR-9994过表达促进增殖相关基因PCNA、CCND1、Bcl-2表达，抑制凋亡基因c-PARP表达，促进EMT相关基因N-cadherin、MMP7、Twist和Vimentin表达及抑制E-cadherin表达；敲低则作用相反。
结论 piR-9994异常表达影响胃癌细胞增殖、侵袭、转移及EMT进程。piR-9994可能是胃癌新的标志物和治疗靶点。

Abstract:
Objective To investigate the effect of piR-9994 on the biological behavior of gastric cancer cells and its possible mechanism.
Methods The expression of piR-9994 in gastric cancer cell lines (MGC803 and AGS) and normal gastric epithelial cells (GES-1) were detected by qRT-PCR. MGC803 cell line with piR-9994 overexpression and knockdown were constructed. The effects of piR-9994 expression changes on cell proliferation were detected by MTT and clone formation assay. The scratch wound healing assay and Transwell invasion assay were used to detect cell migration and invasion abilities. qRT-PCR and Western blot were used to detect cell proliferation and EMT-related genes expression.
Results The expression level of piR-9994 in MGC803 cells was significantly higher than that in normal gastric epithelial cell line GES-1 (P < 0.05). The overexpression of piR-9994 promoted the proliferation, invasion and metastasis of gastric cancer cells, while piR-9994 knockdown had the opposite effect. piR-9994 expression was closely related to cell cycle, especially in S phase. The overexpression of piR-9994 promoted the expression of proliferation-related genes PCNA, CCND1 and Bcl-2, inhibited the expression of apoptosis gene C-PARP, and promoted the expression of EMT-related genes N-cadherin, MMP7, Twist and Vimentin; while piR-9994 knockdown had the opposite effect.
Conclusion Abnormal expression of piR-9994 affects the proliferation, invasion, metastasis and EMT process of gastric cancer cells. piR-9994 may be a new biomarker and therapeutic target for gastric cancer.

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