黄夹次甙乙抑制胃癌MKN-45细胞增殖作用及机制探讨  

Inhibitory effect and mechanism of neriifolin on the proliferation of gastric cancer MKN-45 cells

作　　者：张斌 马英俊 伦俊杰 赵萍萍 刘伟 ZHANG Bin;MA Yingjun;LUN Junjie;ZHAO Pingping;LIU Wei(Oncology Department,Changle County People's Hospital,Weifang,Shandong 261000,China;Tuberculosis Prevention and Treatment Department,Changle County Center for Disease Control and Prevention,Weifang,Shandong 261000,China)

机构地区：[1]昌乐县人民医院肿瘤科,山东潍坊261000 [2]昌乐县疾病预防控制中心结核防治科,山东潍坊261000

出　　处：《社区医学杂志》2025年第3期83-89,共7页Journal Of Community Medicine

基　　金：潍坊市卫生健康委员会中医药研究项目(WFZYY2022-1-002)。

摘　　要：目的探究黄花夹竹桃有效成分黄夹次甙乙对胃癌MKN-45细胞增殖的影响及可能的作用机制。方法以不同浓度的黄夹次甙乙(0、5、10和20μmol/L)处理胃癌MKN-45细胞,利用细胞计数8(CCK-8)实验检测各组细胞的增殖情况;利用5-乙炔基-2'-脱氧尿苷(EDU)实验检测各组细胞DNA合成情况;利用克隆平板实验检测各组细胞克隆形成能力;利用网络药理学方法预测黄夹次甙乙的直接作用靶点;利用蛋白质印迹实验检测黄夹次甙乙对其直接作用靶点热休克蛋白90α家族A类成员1(HSP90AA1)的表达影响。构建阴性对照和HSP90AA1敲低的细胞,利用CCK-8实验探究黄夹次甙乙对两者的抑制差异。结果与0μmol/L处理组增殖率(100.00±8.56)%相比,5、10和20μmol/L处理组细胞相对增殖率分别为(83.56±4.41)%、(73.27±4.75)%和(47.37±6.20)%,F=37.992,t值分别为6.473、9.739和14.700,P值分别为0.023、0.010和0.004;与0μmol/L处理组EDU阳性率(40.10±4.67)%相比,5、10和20μmol/L处理组细胞EDU阳性率分别为(25.45±4.71)%、(14.98±3.51)%和(7.15±2.85)%,F=37.853,t值分别为5.387、12.420和20.041,P值分别为0.032、0.006和<0.001;与0μmol/L处理组克隆平板数(418.00±33.41)个相比,5、10和20μmol/L处理组细胞克隆形成数分别为(294.00±33.72)、(117.30±13.05)和(81.00±13.75)个,F=113.850,t值分别为6.369、39.901和42.463,P值分别为0.023、<0.001和<0.001。通过在TargetNet和Super-PERD网站进行网络药理学分析,结果显示HSP90AA1是黄夹次甙乙的直接作用靶点。蛋白质印迹结果显示,与0μmol/L处理组相比,各处理组细胞的HSP90AA1的蛋白表达水平均减少,F=68.127,t值分别为6.309、20.393和24.477,P值分别为0.024、0.002和<0.001。此外,通过构建HSP90AA1敲低的细胞发现,相比于阴性对照组细胞的抑制率(51.65±8.76)%,20μmol/L黄夹次甙乙对HSP90AA1敲低细胞的抑制率减少,为(27.62±6.81)%,t=3.755,P=0.018。结论黄夹次甙乙抑制胃癌MKN-45细胞增殖Objective To investigate the effects of neriifolin,an active component of Thevetia peruviana,on the proliferation of gastric cancer MKN-45 cells and its potential mechanism of action.Methods Gastric cancer MKN-45 cells were treated with different concentrations of neriifolin(0,5,10,and 20 μmol/L).Cell proliferation was assessed using the Cell counting kit-8(CCK-8) assay.DNA synthesis was evaluated using the 5-ethynyl-2'-deoxyuridine(EdU) incorporation assay.The colony formation ability of the cells was examined using a colony formation assay.Potential direct targets of neriifolin were predicted using network pharmacology tools such as TargetNet and Super-PERD.The expression levels of the direct target,Heat shock protein 90 alpha family class A member 1(HSP90AA1),were analyzed by western blotting.Negative control and HSP90 AA1-knockdown cells were constructed to assess the differences in inhibitory effects of neriifolin using the CCK-8 assay.Results Compared with the proliferation rate in the 0 μmol/L treatment group(100.00±8.56)%,the relative proliferation rates in the 5,10,and 20 μmol/L neriifolin-treated groups were significantly reduced to(83.56±4.41%),(73.27±4.75%),and(47.37±6.20%),respectively(F value was 37.992;t value were 6.473,9.739 and 14.700;P value were 0.023,0.010 and 0.004).Similarly,compared with the EdU-positive rate in the 0 μmol/L treatment group(40.10±4.67%),the EdU-positive rates in the 5,10,and 20 μmol/L treatment groups were significantly reduced to(25.45±4.71%),(14.98±3.51%),and(7.15±2.85%),respectively(F value was37.853;t value were 5.387,12.420 and 20.041;P value were 0.032,0.006 and <0.001).The number of colonies in the colony formation assay also significantly decreased in the 5,10,and 20 μmol/L groups(294.00±33.72,117.30±13.05,and 81.00±13.75,respectively) compared with the 0 μmol/L group(418.00±33.41)(F value was 113.850;t value were 6.369,39.901 and 42.463;P value were 0.023,<0.001 and <0.001).Network pharmacology analysis through TargetNet and Super-PERD indicated

关 键 词：胃癌 黄夹次甙乙 增殖 热休克蛋白90α家族A类成员1 

分 类 号：R735.2[医药卫生—肿瘤]