异牡荆素通过miR-339-5p/HSPA8轴调节胰腺癌细胞的生物学行为  

Isovitexin regulates proliferation,migration and invasion of pancreatic cancer cells via the miR-339-5p/HSPA8 axis

作　　者：闫玲新 李森 郭改莉 孟婉秋 徐超[1] YAN Lingxin;LI Sen;GUO Gaili;MENG Wanqiu;XU Chao(First Ward of Gastroenterology Department,Handan Central Hospital,Handan 056000,China)

机构地区：[1]邯郸市中心医院消化内科一病区,056000

出　　处：《天津医药》2025年第3期230-235,共6页Tianjin Medical Journal

基　　金：邯郸市科学技术研究与发展计划项目(23422083233)。

摘　　要：目的探究异牡荆素(Isov)对胰腺癌细胞的生物学行为的影响及作用机制。方法采用Isov处理人正常胰腺导管上皮细胞HPDE和PC细胞系;CCK-8检测细胞存活率并计算半数抑制浓度(IC50)。将PC细胞系PANC-1细胞依次分为Control组、Isov组、Isov+in-miR-NC组、Isov+in-miR-339-5p组、Isov+in-miR-339-5p+si-NC组和Isov+inmiR-339-5p+si-HSPA8组。CCK-8、划痕愈合实验和Transwell实验检测PANC-1细胞存活、迁移和侵袭情况;实时荧光定量PCR检测PANC-1细胞中miR-339-5p和热休克蛋白家族A成员8(HSPA8)mRNA表达情况;Western blot法检测各组细胞中蛋白HSPA8的表达情况;双萤光素酶报告基因检测miR-339-5p与HSPA8的靶向作用;异种移植裸鼠模型验证Isov的体内抗癌作用。结果Isov可抑制PC细胞增殖,但对HPDE细胞几乎没有细胞毒性。Isov可显著降低PANC-1细胞存活率和划痕愈合率,减少侵袭细胞数量,上调miR-339-5p表达,下调HSPA8 mRNA和蛋白水平(P<0.05),而这种作用可被下调miR-339-5p阻断(P<0.05)。HSPA8是miR-339-5p的靶标基因,敲低HSPA8后逆转了Isov对PANC-1细胞恶性生物学行为的调控作用(P<0.05)。体内研究证实,Isov治疗后,裸鼠肿瘤体积和质量减小,且miR-339-5p表达升高,HSPA8 mRNA表达降低(P<0.05)。结论Isov可能通过miR-339-5p/HSPA8轴抑制PC细胞的增殖、迁移和侵袭。Objective To explore the biological behavior and mechanism of isovitexin(Isov)on pancreatic cancer cells.Methods Isov was used to treat the human normal pancreatic ductal epithelial cells HPDE and PC cell lines,and CCK-8 was used to detect the cell proliferation and calculate the half inhibitory concentration(IC50).The PC cell line PANC-1 cells were grouped into the control group,the Isov group,the Isov+in-miR-NC group,the Isov+in-miR-339-5p group,the Isov+in-miR-339-5p+si-NC group and the Isov+in-miR-339-5p+si-HSPA8 group.The survival,migration and invasion of PANC-1 cells were detected by CCK-8,scratch healing assay and Transwell assay.Real time fluorescence quantitative PCR was used to detect the mRNA expression of miR-339-5p and heat shock protein family A member 8(HSPA8)in PANC-1 cells.Western blot assay was used to detect protein HSPA8 expression in various groups of cells.Dual luciferase reporter gene was used to detect the targeting effect of miR-339-5p and HSPA8.A xenograft nude mouse model was used to determine the in vivo anticancer effects of Isov.Results Isov inhibited PC cell proliferation but had little cytotoxicity to HPDE cells.Isov could obviously reduce the survival rate and scratch healing rate of PANC-1 cells,reduce the number of invasive cells,up-regulate miR-339-5p expression and down-regulate HSPA8 mRNA and protein levels(P<0.05),while these effects were blocked by down-regulated miR-339-5p(P<0.05).In addition,HSPA8 was the target gene of miR-339-5p,and knockdown of HSPA8 reversed the regulatory effect of Isov on the malignant biological behavior of PANC-1 cells.In vivo studies confirmed that after Isov treatment,the tumor volume and weight of nude mice decreased,the expression of miR-339-5p was increased and the expression of HSPA8 mRNA was decreased(P<0.05).Conclusion Isov may inhibit the proliferation,migration and invasion of PC cells through the miR-339-5p/HSPA8 axis.

关 键 词：牡荆素 胰腺肿瘤 细胞增殖 HSPA8 miR-339-5p 

分 类 号：R735.9[医药卫生—肿瘤]