Lv Haidong, Zhou Dixia, Liu Guoqing
Department of Tumor Surgery, Qinghai People's Hospital, Xining 810007, Qinghai, China.
Department of Tumor Surgery, Qinghai People's Hospital, Republic Road No. 2, Xining 810007, Qinghai, China.
Open Med (Wars). 2023 Jan 31;18(1):20220550. doi: 10.1515/med-2022-0550. eCollection 2023.
Long non-coding RNA plasmacytoma variant translocation 1 (PVT1) has been reported to be a vital modulator in tumorigenesis of gastric cancer (GC). However, the detailed regulatory mechanism of PVT1 in GC remains largely unclear. In this work, the expressions of PVT1 and microRNA-16 (miR-16) were detected by quantitative real-time PCR (qRT-PCR) in GC tissues and cell lines. GC cell lines NCI-N87 and MKN45 cell lines were chosen for the following assays. After PVT1 was overexpressed or depleted, CCK-8 and Transwell assays were performed to examine the cell viability and invasive capacity. Cell cycle was analyzed by flow cytometry. The expression of cyclin D1 (CCND1) at mRNA and protein levels was measured by qRT-PCR and western blot. The competitive endogenous RNA molecular mechanism among PVT1, miR-16 and CCND1 was verified by bioinformatics analysis, luciferase-reporter gene assay and RNA immunoprecipitation assay. In the present study, it was revealed that PVT1 expression was remarkably evaluated in GC tissues and cell lines than that in the corresponding control group. PVT1 positively regulated the proliferation, migration and cell cycle progression of GC cells. Besides, miR-16 was identified as a target of PVT1, and CCND1 was identified as a target of miR-16. The depletion of PVT1 promoted the expression of miR-16 and suppressed CCND1 expression. Moreover, either miR-16 inhibitor or CCND1 overexpression plasmid could reverse the promoting effects of PVT1 on the malignant biological behaviors of GC cells. In conclusion, PVT1 promoted CCND1 expression by negatively regulating miR-16 expression to enhance the viability, invasion and cell cycle progression of GC cells.
长链非编码RNA浆细胞瘤变异易位1(PVT1)据报道是胃癌(GC)肿瘤发生中的一个重要调节因子。然而,PVT1在GC中的详细调控机制仍不清楚。在本研究中,通过定量实时PCR(qRT-PCR)检测了GC组织和细胞系中PVT1和微小RNA-16(miR-16)的表达。选择GC细胞系NCI-N87和MKN45细胞系进行后续实验。在PVT1过表达或缺失后,进行CCK-8和Transwell实验以检测细胞活力和侵袭能力。通过流式细胞术分析细胞周期。通过qRT-PCR和蛋白质印迹法检测细胞周期蛋白D1(CCND1)在mRNA和蛋白质水平的表达。通过生物信息学分析、荧光素酶报告基因实验和RNA免疫沉淀实验验证了PVT1、miR-16和CCND1之间的竞争性内源RNA分子机制。在本研究中,发现PVT1在GC组织和细胞系中的表达明显高于相应的对照组。PVT1正向调节GC细胞的增殖、迁移和细胞周期进程。此外,miR-16被鉴定为PVT1的靶标,CCND1被鉴定为miR-16的靶标。PVT1的缺失促进了miR-16的表达并抑制了CCND1的表达。此外,miR-16抑制剂或CCND1过表达质粒均可逆转PVT1对GC细胞恶性生物学行为的促进作用。总之,PVT1通过负向调节miR-16的表达促进CCND1的表达,从而增强GC细胞的活力、侵袭能力和细胞周期进程。