咪达唑仑调节Notch/Snail信号通路对结直肠癌细胞增殖、迁移和上皮间质转化的影响  

Effects of midazolam on the proliferation,migration,and epithelial mesenchymal transition of colorectal cancer cells by regulating the Notch/Snail signaling pathway

作　　者：文竹[1] 李军[1] WEN Zhu;LI Jun(Department of Anesthesiology,Mianyang Central Hospital,Sichuan,Mianyang 621000,China)

机构地区：[1]四川省绵阳市中心医院麻醉科,621000

出　　处：《河北医药》2025年第1期11-16,共6页Hebei Medical Journal

基　　金：发育与再生四川省重点实验室研究基金(编号:SYS20-01)。

摘　　要：目的 探讨咪达唑仑(Mid)对结直肠癌(CRC)细胞HT29增殖、迁移和上皮间质转化(EMT)的影响及其对Notch/锌指蛋白转录因子(Snail)信号通路的调节作用。方法 体外培养HT29细胞并使用0~20μmol/L的Mid处理细胞,筛选最佳浓度,将HT29细胞分为对照组(CT组)、Mid L组(5μmol/L)、Mid M组(10μmol/L)、Mid H组(20μmol/L)和Mid H+Notch激活剂组(midH+VPA-d4组,50μmol/L);平板克隆法检测细胞增殖;细胞划痕实验检测迁移;Transwell实验测定侵袭;流式细胞仪检测凋亡;免疫荧光法检测神经钙黏附蛋白(N-cadherin)、上皮型钙黏素(E-cadherin)蛋白表达;Western blot检测Notch1、Snail蛋白表达。结果 与CT组比较,Mid L、M、H组细胞克隆形成数、迁移率、侵袭细胞数及Notch1、Snail、N-cadherin蛋白表达下降,凋亡率、E-cadherin蛋白表达升高,且呈浓度依赖性(P<0.05);与Mid H组比较,Mid H+VPA-d4组克隆形成数、迁移率、侵袭细胞数及Notch1、Snail、N-cadherin蛋白表达升高,凋亡率、E-cadherin蛋白表达降低(P<0.05)。结论 Mid可能通过抑制Notch/Snail信号通路抑制CRC细胞的增殖、迁移和EMT。Objective To investigate the effects of midazolam(Mid)on the proliferation,migration,and epithelial mesenchymal transition(EMT)of colorectal cancer(CRC)cells HT29 and its regulatory effect on the Notch/zinc finger protein transcription factor(Snail)signaling pathway.Methods HT29 cells were cultured in vitro and treated with 0-20μmol/L Mid to select the optimal concentration.HT29 cells were treated with blank control,low-dose(5μmol/L),medium-dose(10μmol/L)and high-dose Mid(20μmol/L),and high-dose Mid+Notch activator VPA-d4(20μmol/L Mid+50μmol/L VPA-d4).Cell proliferation,migration,invasion and apoptosis were measured by colony formation assay,wound healing assay,Transwell assay and flow cytometry,respectively.Immunofluorescence assay was applied to detect the protein expressions of N-cadherin and E-cadherin.Western blot was applied to detect the protein expressions of Notch1 and Snail.Results Compared with those of blank control,Mid treatment significantly decreased colony number,migratory rate,invasive cell number and protein expressions of Notch1,Snail and N-cadherin,but increased apoptotic rate and protein level of E-cadherin in a dose-dependent manner(P<0.05).Compared with those induced with high-dose Mid,treatment of high-dose Mid+VPA-d4 significantly increased colony number,migratory rate,invasive cell number and protein expressions of Notch1,Snail and N-cadherin,but decreased apoptotic rate and protein level of E-cadherin(P<0.05).Conclusion Mid may inhibit the proliferation,migration,and EMT of CRC cells by inhibiting the Notch/Snail signaling pathway.

关 键 词：咪达唑仑 结直肠癌细胞 Notch/锌指蛋白转录因子(Snail)信号通路 增殖 迁移 

分 类 号：R735.35[医药卫生—肿瘤]