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槐耳颗粒对索拉非尼耐药的肝细胞癌细胞凋亡的影响研究

Effect of Huaier granule on apoptosis of sorafenib resistant hepatocellular carcinoma cells
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摘要 目的探讨槐耳颗粒对索拉非尼耐药的肝细胞癌(HCC)细胞生长的抑制作用及其分子机制。方法利用梯度浓度槐耳颗粒处理HCC细胞,分析槐耳颗粒对索拉非尼耐药的HCC细胞增殖、迁移和侵袭的效果。采用生物信息学手段分析微小RNA-31-5p(miR-31-5p)与sprouty相关EVH1域蛋白1(SPRED1)的可能互作关系。采用实时荧光定量聚合酶链反应(qRT-PCR)检测HCC细胞中miR-31-5p、SPRED1的表达;采用CCK-8、克隆形成、划痕愈合、Transwell以及流式细胞术实验分别检测细胞活力、增殖、迁移、侵袭和凋亡;采用RNA免疫沉淀(RIP)实验和双荧光素酶实验验证miR-31-5p与SPRED1的结合关系。结果槐耳颗粒可以显著抑制索拉非尼耐药的HCC细胞增殖、迁移和侵袭,并诱导细胞凋亡。生物信息学分析发现miR-31-5p在HCC中高表达,而槐耳颗粒可以下调miR-31-5p的表达,抑制索拉非尼耐药的HCC细胞的增殖和转移,并诱导细胞凋亡;miR-31-5p可靶向抑制SPRED1的表达。SPRED1在HCC中表达下调,过表达SPRED1可以逆转miR-31-5p过表达对索拉非尼耐药的HCC增殖和转移的促进作用。结论槐耳颗粒通过miR-31-5p/SPRED1轴抑制索拉非尼耐药的HCC转移,表明槐耳颗粒有潜力作为治疗HCC的新型药物。 Objective To investigate the inhibitory effect and molecular mechanism of Huaier granule on the growth of sorafenib resistant hepatocellular carcinoma(HCC)cells.Methods The gradient concentration of Huaier granule was used to treat HCC cells,and the effect of Huaier granule on the proliferation,migration and invasion of sorafenib resistant HCC cells was analyzed.Bioinformatics methods were used to analyze the possible interaction between microRNA-31-5p(miR-31-5p)and sprouty-related proteins with an EVH1 domain 1(SPRED1).The expression levels of miR-31-5p and SPRED1 in HCC cells were detected by real time fluorescence quantitative polymerase chain reaction(qRT-PCR);cell viability,proliferation,migration,invasion and apoptosis were detected by CCK-8,colony formation,scratch healing,Transwell and flow cytometry;RNA immunoprecipitation(RIP)assay and dual luciferase assay were used to verify the binding relationship between miR-31-5p and SPRED1.Results Huaier granule could significantly inhibit the proliferation,migration and invasion of sorafenib resistant HCC cells,and induce apoptosis.Bioinformatics analysis showed that miR-31-5p was highly expressed in HCC,and Huaier granule was able to down-regulate the expression of miR-31-5p,inhibit the proliferation and metastasis of sorafenib resistant HCC cells,and induce apoptosis;miR-31-5p showed a targeted inhibition effect on the expression of SPRED1.SPRED1 was down-regulated in HCC,and overexpression of SPRED1 was able to reverse the promoting effect of overexpression of miR-31-5p on proliferation and metastasis of sorafenib resistant HCC.Conclusion Huaier granule can inhibit sorafenib resistant HCC metastasis through the miR-31-5p/SPRED1 axis,indicating that Huaier granule has the potential to be used as a novel drug for HCC treatment.
作者 王利兵 于敬坤 曲凤智 程大铭 刘晓刚 WANG Libing;YU Jingkun;QU Fengzhi;CHENG Daming;Liu Xiaogang(the Second Department of Hepatobiliary Surgery,Tangshan City Workers′Hospital in Hebei Province,Tangshan,Hebei,063000)
出处 《实用临床医药杂志》 CAS 2024年第5期44-52,共9页 Journal of Clinical Medicine in Practice
基金 2021年度河北省中医药类科研计划项目(2021413)。
关键词 槐耳颗粒 微小RNA-31-5p sprouty相关EVH1域蛋白1 索拉非尼 耐药性 肝细胞癌 凋亡 Huaier granule microRNA-31-5p sprouty-related proteins with an EVH1 domain 1 sorafenib drug resistance hepatocellular carcinoma apoptosis
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