PDF(Pubmed)
Abstract：
UNASSIGNED: Circular RNAs (circRNAs) are a class of non-coding RNAs which function as novel regulators in human cancers. In this study, we aimed to investigate the functional roles and related molecular mechanisms of circ_0006282 in gastric cancer (GC) progression.
UNASSIGNED: Fifty-five GC patients were enrolled in this study. GC cells (AGS and HGC-27) and normal cells (GES-1) were cultured in RPMI1640 added with 10% FBS and 1% penicillin-streptomycin. Quantitative real-time polymerase chain reaction (qRT-PCR) assay was used to determine the expression levels of circ_0006282, transcription elongation factor B subunit 1 (TCEB1) mRNA, miR-144-5p and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein β (YWHAB; also known as 14-3-3β). RNase R assay was used to determine the characteristic of circ_0006282. Cell Counting Kit-8 (CCK-8) assay and colony formation assay were employed for cell proliferation. Transwell assay was conducted for cell migration and invasion. Western blot assay was carried out to measure the protein levels of Cyclin D1, matrix metalloprotein 9 (MMP9) and YWHAB. Dual-luciferase reporter assay, RNA pull-down assay and RIP assay were adopted to analyze the interaction between miR-144-5p and circ_0006282 or YWHAB. Murine xenograft model assay was performed to explore the function of circ_0006282 in vivo.
UNASSIGNED: Circ_0006282 level was increased in GC tissues and cells compared to normal tissues and cells. Silencing of circ_0006282 restrained GC cell proliferation, migration and invasion. For mechanism analysis, circ_0006282 was identified to function as the sponge for miR-144-5p to positively regulate YWHAB expression in GC cells. Moreover, miR-144-5p inhibition or YWHAB overexpression effectively reversed the impacts of circ_0006282 knockdown on GC cell growth and motility. Additionally, circ_0006282 knockdown blocked tumor growth of GC in vivo.
UNASSIGNED: Circ_0006282 facilitated the malignant behaviors of GC cells through circ_0006282/miR-144-5p/YWHAB axis.
UNASSIGNED: Fifty-five GC patients were enrolled in this study. GC cells (AGS and HGC-27) and normal cells (GES-1) were cultured in RPMI1640 added with 10% FBS and 1% penicillin-streptomycin. Quantitative real-time polymerase chain reaction (qRT-PCR) assay was used to determine the expression levels of circ_0006282, transcription elongation factor B subunit 1 (TCEB1) mRNA, miR-144-5p and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein β (YWHAB; also known as 14-3-3β). RNase R assay was used to determine the characteristic of circ_0006282. Cell Counting Kit-8 (CCK-8) assay and colony formation assay were employed for cell proliferation. Transwell assay was conducted for cell migration and invasion. Western blot assay was carried out to measure the protein levels of Cyclin D1, matrix metalloprotein 9 (MMP9) and YWHAB. Dual-luciferase reporter assay, RNA pull-down assay and RIP assay were adopted to analyze the interaction between miR-144-5p and circ_0006282 or YWHAB. Murine xenograft model assay was performed to explore the function of circ_0006282 in vivo.
UNASSIGNED: Circ_0006282 level was increased in GC tissues and cells compared to normal tissues and cells. Silencing of circ_0006282 restrained GC cell proliferation, migration and invasion. For mechanism analysis, circ_0006282 was identified to function as the sponge for miR-144-5p to positively regulate YWHAB expression in GC cells. Moreover, miR-144-5p inhibition or YWHAB overexpression effectively reversed the impacts of circ_0006282 knockdown on GC cell growth and motility. Additionally, circ_0006282 knockdown blocked tumor growth of GC in vivo.
UNASSIGNED: Circ_0006282 facilitated the malignant behaviors of GC cells through circ_0006282/miR-144-5p/YWHAB axis.
摘要：
环状RNA(circularRNAs,circRNAs)是一类非编码RNA,其在人类癌症中作为新型调节因子起作用。在这项研究中,我们旨在研究circ_0006282在胃癌（GC）进展中的功能作用和相关分子机制。
■本研究纳入了55例GC患者。在添加有10%FBS和1%青霉素-链霉素的RPMI1640中培养GC细胞(AGS和HGC-27)和正常细胞(GES-1)。定量实时聚合酶链反应（qRT-PCR）测定circ_0006282，转录延伸因子B亚基1（TCEB1）mRNA的表达水平，miR-144-5p和酪氨酸3-单加氧酶/色氨酸5-单加氧酶激活蛋白β(YWHAB;也称为14-3-3β)。RNaseR测定用于确定circ_0006282的特征。细胞计数试剂盒-8(CCK-8)测定和集落形成测定用于细胞增殖。对细胞迁移和侵袭进行Transwell测定。进行蛋白质印迹测定以测量细胞周期蛋白D1，基质金属蛋白9（MMP9）和YWHAB的蛋白质水平。双荧光素酶报告基因测定,采用RNA下拉法和RIP法分析miR-144-5p与circ_0006282或YWHAB的相互作用。进行鼠异种移植模型测定以探索circ_0006282在体内的功能。
■Circ_0006282水平在GC组织和细胞中与正常组织和细胞相比增加。沉默circ_0006282克制GC细胞增殖，移民和入侵。对于机理分析，circ_0006282被鉴定为充当miR-144-5p的海绵以正向调节GC细胞中的YWHAB表达。此外，miR-144-5p抑制或YWHAB过表达有效逆转了circ_0006282敲低对GC细胞生长和运动的影响。此外,circ_0006282敲低阻断了体内GC的肿瘤生长。
■Circ_0006282通过circ_0006282/miR-144-5p/YWHAB轴促进GC细胞的恶性行为。
■本研究纳入了55例GC患者。在添加有10%FBS和1%青霉素-链霉素的RPMI1640中培养GC细胞(AGS和HGC-27)和正常细胞(GES-1)。定量实时聚合酶链反应（qRT-PCR）测定circ_0006282，转录延伸因子B亚基1（TCEB1）mRNA的表达水平，miR-144-5p和酪氨酸3-单加氧酶/色氨酸5-单加氧酶激活蛋白β(YWHAB;也称为14-3-3β)。RNaseR测定用于确定circ_0006282的特征。细胞计数试剂盒-8(CCK-8)测定和集落形成测定用于细胞增殖。对细胞迁移和侵袭进行Transwell测定。进行蛋白质印迹测定以测量细胞周期蛋白D1，基质金属蛋白9（MMP9）和YWHAB的蛋白质水平。双荧光素酶报告基因测定,采用RNA下拉法和RIP法分析miR-144-5p与circ_0006282或YWHAB的相互作用。进行鼠异种移植模型测定以探索circ_0006282在体内的功能。
■Circ_0006282水平在GC组织和细胞中与正常组织和细胞相比增加。沉默circ_0006282克制GC细胞增殖，移民和入侵。对于机理分析，circ_0006282被鉴定为充当miR-144-5p的海绵以正向调节GC细胞中的YWHAB表达。此外，miR-144-5p抑制或YWHAB过表达有效逆转了circ_0006282敲低对GC细胞生长和运动的影响。此外,circ_0006282敲低阻断了体内GC的肿瘤生长。
■Circ_0006282通过circ_0006282/miR-144-5p/YWHAB轴促进GC细胞的恶性行为。
