miR-155介导TGF-β/SMAD-2信号参与胃癌及清热化湿方的干预研究  

Study on the involvement of TGF-β/ Smad-2 signal mediated by miR-155 in gastric cancer and the intervention of Qingrehuashi formula

作　　者：赵俊佳 陈远能[2] 张涛[2] 钟婵 黄李冰雪 敖先伟 ZHAO jun-jia;CHEN Yuan-neng;ZHANG Tao;ZHONG Chan;HUANG Li-bing-xue;AO Xian-wei(Guangxi University of Chinese Medicine,Nanning 530000,China;Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530000,China)

机构地区：[1]广西中医药大学,广西南宁530000 [2]广西中医药大学附属瑞康医院,广西南宁530000

出　　处：《时珍国医国药》2021年第11期2613-2618,共6页Lishizhen Medicine and Materia Medica Research

基　　金：广西自然科学基金(2020GXNSFAA238030)。

摘　　要：目的通过构建稳定沉默miR-155人胃腺癌SGC-7901细胞系,探讨miR-155参与胃癌的生物学行为。观察清热化湿方对胃癌细胞的增殖、周期、侵袭和迁徙,以及TGF-β、SMAD2、CyclinD1的表达水平变化,分析清热化湿方防治胃癌的机制。方法应用基因转染技术构建稳定沉默miR-155胃癌SGC-7901细胞系,观察胃癌细胞的生物学行为变化。应用血清药理学的方法制备含药血清,分为空白对照组(control)、阴性对照组(sh-NC)、sh-miR-155组、治疗组(低、中、高剂量)即sh-miR-155+(5%、10%、20%含药血清),CCK8法检测细胞增殖,流式细胞仪检测细胞周期和凋亡,Transwell法检测细胞的迁徙和侵袭,qRT-PCR、Western-blotting检测细胞TGF-β、SMAD2、CyclinD1 mRNA及蛋白表达水平。结果沉默miR-155可促进胃癌SGC-7901细胞增殖、迁徙和侵袭(P<0.05),促进TGF-β、SMAD2、CyclinD1 mRNA及蛋白表达(P<0.05);清热化湿方通过上调miR-155表达,诱导胃癌SGC-7901细胞的凋亡,抑制胃癌SGC-7901细胞增殖、迁徙和侵袭(P<0.05);并且抑制胃癌SGC-7901细胞TGF-β、SMAD2、CyclinD1 mRNA及蛋白表达(P<0.05)。结论稳定沉默miR-155可上调TGF-β、SMAD2、CyclinD1表达,促进胃癌进展。清热化湿方可上调miRNA155的表达,抑制TGF-β、SMAD2、CyclinD1的表达,诱导细胞凋亡,抑制细胞增殖、迁徙和侵袭,达到协同防治胃癌的效用。Objective To investigate the role of miR-155 in the biological behavior of gastric cancer, we constructed a stable silenced human gastric adenocarcinoma SGC-7901 cell line.To observe the proliferation, cycle, invasion and migration of gastric cancer cells as well as the changes in the expression levels of TGF-β, Smad2 and CyclinD1, and analyze the mechanism of Qingrehuashi formula in the prevention and treatment of gastric cancer.Methods Stably silenced miR-155 gastric cancer cell line SGC-7901 was constructed by gene transfection technique, and the changes of biological behavior of gastric cancer cells were observed.Serum pharmacological method was used to prepare pharmaceutical clear, which was divided into control group, negative control group(sh-NC), sh-miR-155 group, treatment group(low, medium and high dose) namely sh-miR-155+(5%, 10% and 20% containing pharmaceutical clear). Cell proliferation was detected by CCK8 assay, cell cycle and apoptosis were detected by flow cytometry, and cell migration and invasion were detected by Transwell assay.MRNA and protein expression levels of TGF-β, Smad2 and CyclinD1 were detected by qRT-PCR and Western-blotting.Results Silencing miR-155 promoted the proliferation, migration and invasion of SGC-7901 cells(P < 0.05), and promoted the mRNA and protein expressions of TGF-β, Smad2 and CyclinD1(P < 0.05).Qingrehuashi formula induced apoptosis of SGC-7901 cells by up-regulating the expression of miR-155, and inhibited proliferation, migration and invasion of SGC-7901 cells(P< 0.05).Moreover, the mRNA and protein expressions of TGF-β, Smad2 and CyclinD1 in gastric cancer SGC-7901 cells were inhibited(P < 0.05).Conclusion Stably silencing miR-155 can up-regulate the expressions of TGF-β, Smad2 and CyclinD1, and promote the progression of gastric cancer.Qingrehuashi formula can up-regulate the expression of miRNA155, inhibit the expression of TGF-β, Smad2 and CyclinD1, induce cell apoptosis, inhibit cell proliferation, migration and invasion, and achieve synergistic effic

关 键 词：清热化湿方 microRNA155 TGF-Β SMAD2 胃癌 

分 类 号：R284.2[医药卫生—中药学]