败酱草水提物通过调控circ_0000936/miR-665抑制胶质瘤细胞增殖、迁移及侵袭  

Water extract from patrinia scabiosaefolia inhibits the proliferation,migration and invasion of glioma cells by regulating circ_0000936/miR-665

作　　者：张正平[1] 李坤正[1] 吴彬杰 刘熹 巨虎 张晓璐 ZHANG Zhengping;LI Kunzheng;WU Binjie;LIU Xi;JU Hu;ZHANG Xiaolu(Department of Neurosurgery,Affiliated Hospital of Qinghai University,Qinghai Xining 810000,China;Department of Neurosurgery,Wuxi People's Hospital Affiliated to Nanjing Medical University,Jiangsu Wuxi 214023,China.)

机构地区：[1]青海大学附属医院神经外科,青海西宁810000 [2]南京医科大学附属无锡市人民医院神经外科,江苏无锡214023

出　　处：《现代肿瘤医学》2024年第6期1017-1024,共8页Journal of Modern Oncology

基　　金：青海大学青年科研基金项目(编号:2020-QYY-6)。

摘　　要：目的:探讨败酱草水提物对胶质瘤细胞生物行为的影响及其可能作用机制。方法:收集2019年2月至2020年1月期间于本院接受手术切除并经病理证实的31例胶质瘤组织,qRT-PCR法检测正常脑组织与胶质瘤组织中circ_0000936、miR-665的表达量;Pearson法分析胶质瘤组织中circ_0000936和miR-665表达量的相关性;体外培养人胶质瘤细胞LN229,不同浓度的败酱草(0.5 mg/mL、1.0 mg/mL、2.0 mg/mL)处理LN229细胞,si-NC、si-circ_0000936分别转染至LN229细胞,pcDNA-circ_0000936转染至LN229细胞后加入2.0 mg/mL败酱草处理细胞;CCK-8、平板克隆形成实验、Transwell小室实验与流式细胞术分别检测细胞增殖、克隆形成、迁移、侵袭及凋亡能力;双荧光素酶报告基因实验检测circ_0000936与miR-665的靶向关系;Western blot法检测Bax、Bcl-2蛋白表达量。结果:胶质瘤组织中circ_0000936的表达量高于正常脑组织(P<0.05),而miR-665的表达量低于正常脑组织(P<0.05);circ_0000936和miR-665呈负相关(r=-0.980,P<0.001);败酱草能够以浓度依赖性方式降低细胞活力、细胞克隆形成率、Bcl-2蛋白水平和circ_0000936的表达量(P<0.05),迁移及侵袭细胞数减少(P<0.05),而凋亡率、Bax蛋白水平和miR-665的表达量升高(P<0.05);转染si-circ_0000936后miR-665的表达量升高(P<0.05),细胞活力、细胞克隆形成率和Bcl-2蛋白水平降低(P<0.05),迁移及侵袭细胞数减少(P<0.05),凋亡率和Bax蛋白水平升高(P<0.05);转染pcDNA-circ_0000936能够逆转败酱草对LN229细胞生物学行为的作用。结论:败酱草水提物可通过调控circ_0000936/miR-665诱导胶质瘤细胞凋亡,并阻碍增殖、迁移及侵袭。Objective:To explore the effect of water extract from patrinia scabiosaefolia on the biological behavior of glioma cells and its possible mechanism.Methods:Thirty-one cases of glioma tissue from February 2019 to January 2020 who underwent surgical resection and were pathologically confirmed in our hospital were collected.qRT-PCR method was used to detect the expression of circ_0000936 and miR-665 in normal brain tissue and glioma tissue.Pearson method was used to analyze the correlation between circ_0000936 and miR-665 expression in glioma tissues.Human glioma cells LN229 were cultured in vitro,and different concentrations of patrinia scabiosaefolia(0.5 mg/mL,1.0 mg/mL,2.0 mg/mL)were treated with LN229 cells.si-NC and si-circ_0000936 were transfected into LN229 cells respectively.After pcDNA-circ_0000936 was transfected into LN229 cells,2.0 mg/mL patrinia scabiosaefolia was added to treat the cells.CCK-8,plate clone formation experiment,Transwell chamber experiment and flow cytometry were used to detect cell proliferation,colony formation,migration,invasion and apoptosis.The dual-luciferase reporter assay was used to detect the targeting relationship between circ_0000936 and miR-665.Western blot method was used to detect the expression of Bax and Bcl-2 protein.Results:The expression of circ_0000936 in glioma tissue was higher than that in normal brain tissue(P<0.05),while the expression of miR-665 was lower than that in normal brain tissue(P<0.05).circ_0000936 and miR-665 were negatively correlated(r=-0.980,P<0.001).Patrinia scabiosaefolia could reduce cell viability,clone formation rate and could reduce the protein level of Bcl-2 and the expression of circ_0000936 in a concentration-dependent manner(P<0.05),and the number of migration and invasion cells was reduced(P<0.05),while the rate of apoptosis,the protein level of Bax and the expression of miR-665 were increased(P<0.05).After transfection with si-circ_0000936,the expression of miR-665 was increased(P<0.05),and cell viability,clone formation rate and the p

关 键 词：败酱草 胶质瘤 circ_0000936 miR-665 细胞增殖 迁移 侵袭 

分 类 号：R730.264[医药卫生—肿瘤]