文章摘要
叶晓荣,李修靖,刘铭宇,等.新鱼腥草素钠抑制胰腺癌 Panc-1细胞恶性生物学行为及其作用机制研究[J].安徽医药,2024,28(7):1302-1306.
新鱼腥草素钠抑制胰腺癌 Panc-1细胞恶性生物学行为及其作用机制研究
Effects of sodium new houttuyfonate on the malignant biological behavior of pancreatic cancer cells Panc-1 and its underlying mechanism
  
DOI:10.3969/j.issn.1009-6469.2024.07.007
中文关键词: 鱼腥草属  细胞凋亡  新鱼腥草素钠  胰腺癌  细胞增殖  肿瘤侵润  迁移  上皮间质转化
英文关键词: Houttuynia  Apoptosis  Sodium new houttuyfonate  Pancreatic cancer  Cell Proliferation  Migration  Neoplasm
基金项目:
作者单位E-mail
叶晓荣 安徽中医药大学第一附属医院脾胃科安徽合肥 230031  
李修靖 安徽中医药大学第一附属医院脾胃科安徽合肥 230031  
刘铭宇 安徽中医药大学第一附属医院脾胃科安徽合肥 230031  
余跃 安徽中医药大学第一附属医院脾胃科安徽合肥 230031
.中国科学技术大学附属第一医院安徽省立医院消化科安徽合肥 230001 
yuyuemd@ustc.edu.cn 
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中文摘要:
      目的探究新鱼腥草素钠(SNH)对胰腺癌 Panc-1细胞增殖、侵袭及迁移能力的影响及其潜在作用机制。方法自 2023年 3月至 2024年 2月,取对数期人胰腺癌 Panc-1细胞,分为对照组(无菌双蒸水)、各不同梯度 SNH处理组( 100 μmol/L组、 200 μmol/L组、 400 μmol/L组、 600 μmol/L组及 800 μmol/L组)。细胞计数试剂( CCK-8)实验检测 Panc-1细胞活率;细胞划痕及 Transwell实验分别检测细胞迁移及侵袭能力;实时荧光定量聚合酶链式反应( qRT-PCR)检测细胞基因水平的变化。蛋白质印迹法检测凋亡蛋白胱天蛋白酶 -3(caspase-3)、多聚二磷酸腺苷核糖聚合酶( PARP)及上皮间质转化( EMT)相关蛋白神经钙黏素( N-cadherin,CDH)、闭合蛋白( occludin,OCLN)的表达。结果 CCK-8实验结果显示, 24 h时, 100、200及 400 μmol/L组细胞活率与对照组相比差异无统计学意义; 600、800 μmol/L组显著降低( P=0.002,P<0.001); 48、72 h时, 100、200 μmol/L组细胞活率未见降低, 400、600、800 μmol/L组明显下降( P<0.001),各组 48 h细胞活率分别为( 62.51±9.59)%、(36.94±5.06)%及( 17.94±0.56)%,72 h降至( 46.00±3.04)%、(22.93±4.75)%及( 5.80±0.42)%。细胞划痕结果显示, 100 μmol/L组 24 h细胞迁移率即下降(P<0.05)48 h迁移率( 4.37±1.92)%及 72 h迁移率( 3.75±2.47)%均明显低于对照组( P<0.01); 200 μmol/L组细胞迁移率同样下降, 72 h率为( 6.25±5.16)%;Transwell实验结果显示,与对照组相比, 24 h时 200、400 μmol/L组侵袭细胞数显著减少(均 P<0.001)迁移; qRT-PCR结果显示, 400 μmol/L组 CDH2基因表达下调( P<0.05),OCLN表达上调( P<0.05);蛋白质印迹法检测, 400 μmol/L组 caspase-3、PARP表达降低( P<0.05)cleaved caspase-3升高( P<0.05)。 200、400 μmol/L组 CDH表达降低( P<0.001), OCLN升高( P<0.001)。结论 SNH抑制胰腺癌细胞的增殖、迁移和侵袭,可能与 SNH促进细胞凋亡,抑制 EMT进程有关。
英文摘要:
      Objective To investigate the effect of sodium new houttuyfonate (SNH) on the proliferation, migration and invasion of pancreatic cancer cells Panc-1 and its potential mechanism.Methods From March 2023 to February 2024, Panc-1 cells in logarithmic phase were assigned into control group (sterile distillation-distillation water), SNH treatment groups with different concentrations (100μmol/L group, 200 μmol/L group, 400 μmol/L group, 600 μmol/L group and 800 μmol/L group). The cell viability of Panc-1 was detected by Cell Counting Kit-8 (CCK-8) assay. Cell migration and invasion ability were detected by cell scratch and Transwell assay respectively. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the changes of gene level in cells. The expression of apoptosis protein caspase-3, poly (ADP-ribose) polymerase (PARP) and epithelial mesenchymal transformation(EMT) related proteins, CDH and OCLN were detected by Western blot.Results The CCK-8 results demonstrated that there was no significant difference in cell viability between the 100, 200 and 400 μmol/L group and the control group at 24 h while it was significantly decreased in 600 and 800 μmol/L group (P = 0.002, P < 0.001). There was no decrease in cell viability in 100 and 200 μmol/Lgroups at 48 and 72 h while it was significantly decreased in 400, 600 and 800 μmol/L groups (P <0.001); the cell viabilities in 400,600 and 800 μmol/L groups at 48 h were (62.51±9.59) %, (36.94±5.06) % and (17.94±0.56) %, respectively, and decreased to (46.00±3.04) %, (22.93±4.75) % and (5.80±0.42) % at 72 h. Cell scratch results showed that cell mobility decreased at 24 h in the 100 μmol/Lgroup (P < 0.05), the mobility at 48 h [(4.37±1.92) %] and 72 h [(3.75±2.47) %] were significantly lower than those of control group (P < 0.01). The cell mobility of 200 μmol/L group also decreased, and the mobility of 72 h was (6.25±5.16)%. Transwell experiment results showed that compared with the control group, the number of invasive cells in 200 and 400 μmol/L groups decreased significantlyat24 h (P < 0.001); qRT-PCR results revealed that CDH2 gene expression in 400 μmol/L group was down-regulated (P < 0.05), while OCLN gene expression was up-regulated (P < 0.05); Western blot results showed that the expression of caspase-3 and PARP in 400 μmol/L group decreased (P < 0.05), while cleaved caspase-3 increased (P < 0.05). CDH expression decreased (P < 0.001), and OCLN increased in 200 and 400 μmol/L groups (P < 0.001).Conclusion SNH inhibits proliferation, migration and invasion of pancreaticcancer cells, which may be related to the promotion of apoptosis and inhibition of EMT process by SNH.
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