摘要
目的探讨槐角黄酮对结直肠癌SW620细胞增殖和凋亡的影响及分子机制。方法SW620细胞用RPMI-1640培养基常规培养,作为对照组,分别用浓度为2.5 mg/ml、5 mg/ml、10 mg/ml的槐角黄酮处理,记为槐角黄酮-低、中、高组;将miR-4284抑制剂及阴性对照载体质粒转染至SW620细胞,记为anti-miR-4284组、anti-miR-NC组;将miR-4284模拟物及阴性对照转染至SW620细胞后用10 mg/ml的槐角黄酮处理,记为槐角黄酮+miR-4284组、槐角黄酮+miR-NC组。MTT检测细胞抑制率;展开克隆形成实验并检测相关克隆细胞数量;应用流式细胞术确认细胞凋亡情况;蛋白表达水平采用蛋白质印迹法完成;实时荧光定量PCR(RT-qPCR)检测结直肠癌组织和SW620细胞中miR-4284的表达水平。结果不同浓度槐角黄酮处理后,SW620细胞增殖活性明显降低,细胞凋亡率明显升高,克隆形成实验显示克隆细胞形成数量减少,cleaved-caspase 3和cleaved-caspase 9蛋白均呈高表达态势发展,miR-4284表达水平降低,呈浓度依赖性,差异有统计学意义(P<0.05)。抑制miR-4284表达后,SW620细胞增殖抑制活性明显升高,细胞凋亡率升高,细胞克隆形成数减少,cleaved-caspase 3和cleaved-caspase 9蛋白呈高表达态势发展,差异有统计学意义(P<0.05)。miR-4284过表达逆转了槐角黄酮对结直肠癌SW620细胞增殖和凋亡的作用。结直肠癌组织中miR-4284表达水平升高,差异有统计学意义(P<0.05)。结论槐角黄酮通过下调miR-4284表达抑制结直肠癌SW620细胞增殖,促进细胞凋亡。
Objective To observe the effects of flavones in Sophorae Fructus on the proliferation and apoptosis of colorectal cancer SW620 cells and decipher the molecular mechanism.Methods SW620 cells were cultured in the RPMI-1640 medium and assigned into control,low-,medium-,and high-dose(2.5,5,and 10 mg/ml,respectively)fla⁃vones,anti-miR-4284,anti-miR-NC,flavones+miR-4284,and flavones+miR-NC groups.The MTT assay was used to assess the inhibition rate on the cells.The colony formation assay was employed to count the colonies,and apoptosis was examined by flow cytometry.Western blotting was employed to determine the protein levels and real-time fluorescent quantitative PCR(RT-qPCR)to determine the expression of miR-4284 in the colorectal cancer tissue and SW620 cells.Results Different concentrations of flavones in Sophorae Fructus inhibited the proliferation,promoted the apoptosis,and reduced the colony formation of SW620 cells.Furthermore,the flavones up-regulated the protein levels of cleaved-caspase 3 and cleaved-caspase 9 and down-regulated the expression of miR-4284 in a concentration-dependent man⁃ner(P<0.05).After the expression of miR-4284 was inhibited,the SW620 cells showed increased proliferation and ap⁃optosis,decreased colony formation,and high expression of cleaved-caspase 3 and cleaved-caspase 9(P<0.05).The overexpression of miR-4284 reversed the effects of flavones in Sophorae Fructus on the proliferation and apoptosis of SW620 cells.The expression level of miR-4284 was up-regulated in colorectal cancer(P<0.05).Conclusion The flavones in Sophorae Fructus can inhibit the proliferation and promote the apoptosis of colorectal cancer SW620 cells by down-regulating the expression of miR-4284.
作者
张海波
黄宏春
王秀敏
王少芳
ZHANG Hai-bo;HUANG Hong-chun;WANG Xiu-min;WANG Shao-fang(Department of Gastroenterology,The People′s Hospital of Anyang City,Anyang Henan 455000;Department of Re-spiratory Medicine,The People′s Hospital of Anyang City,Anyang Henan 455000)
出处
《世界中西医结合杂志》
2023年第10期1970-1974,共5页
World Journal of Integrated Traditional and Western Medicine
基金
河南省医学科技攻关计划项目(2018021010)。
关键词
槐角黄酮
miR-4284
结直肠癌
增殖
凋亡
Flavones in Sophorae Fructus
miR-4284
Colorectal Cancer
Proliferation
Apoptosis
