摘要
目的 探讨瑞芬太尼(REM)调控微小RNA-21-3p(miR-21-3p)/SMAD家族成员7(SMAD7)信号轴影响甲状腺癌(TC)细胞生长的作用机制。方法 使用0~500 ng/m L的REM处理培养的TC细胞(TPC-1),CCK-8法检测细胞活力,筛选出REM的最佳浓度。将细胞分为对照组(Control组)、REM低、中、高浓度组(REM-L组、REM-M组、REM-H组)、REM+miR-NC(REM-H+miR-NC组),高浓度REM+miR-21-3p minics组(REM-H+miR-21-3p minics组)。qRT-PCR法分析TPC-1细胞中miR-21-3p、SMAD7的表达水平,平板克隆法检测TPC-1细胞增殖。流式细胞仪法检测TPC-1细胞凋亡,双荧光素酶实验检测miR-21-3p与SMAD7的靶向关系,Western blot分析法检测SMAD7蛋白表达。结果 选择浓度5 ng/m L、50 ng/m L、500 ng/m L的REM进行后续实验。TPC-1细胞中miR-21-3p水平比NTHY-ORI3.1细胞高,SMAD7 mRNA水平比NTHY-ORI3.1细胞低(P<0.05)。与Control组比较,REM-L、REM-M、REM-H组细胞克隆形成数、miR-21-3p水平依次降低,细胞凋亡率、SMAD7表达逐渐依次升高(P<0.05);与REM-H+miR-NC组比较,REM-H+miR-21-3p minics组细胞克隆形成数、miR-21-3p表达水平升高,细胞凋亡率、SMAD7表达降低(P<0.05);REM-H组与REM-H+miR-NC组细胞克隆形成数、细胞凋亡率、miR-21-3p、SMAD7表达水平无差异(P> 0.05)。双荧光素酶实验显示,miR-21-3p minics组WT-SMAD7荧光素酶活性比miRNC组降低(P<0.05)。结论 REM抑制miR-21-3p/SMAD7信号轴影响TC细胞生长。
Objective To investigate the role of remifentanil(REM)on the growth of thyroid cancer(TC)cells by regulating the microRNA-21-3p(miR-21-3p)/SMAD family member 7(SMAD7)axis.Methods In vitro cultured TC cells(TPC-1)were treated with REM at varying concentrations ranging from 0ng/mL to 500ng/mL,and CCK-8 assay was applied to detect cell viability.The optimal concentration of REM was screened.Cells were induced with blank control,low-dose,medium-dose and high-dose REM,high-dose REM plus transfection of miR-NC or miR-21-3p mimics.Quantitative real-time polymerase chain reaction(QRT-PCR)was applied to detect the expression levels of miR-21-3p and SMAD7 in TPC-1 cells.Cell proliferation and apoptosis were examined by colony formation assay and flow cytometry,respectively.Dua l-luciferase reporter assay was applied to detect the targeting relationship between miR-21-3p and SMAD7.Western blot was applied to detect protein expression of SMAD7.Results REM at concentrations of 5ng/mL,50ng/mL,and 500ng/mL were selected for subsequent experiments.The mRNA level of miR-21-3p in TPC-1 cells was significantly higher than that in NTHY-ORI3.1 cells,and the mRNA level of SMAD7 was significantly lower than that in NTHY-ORI3.1 cells(P<0.05).Compared with the blank control,low-dose,medium-dose and high-dose REM treatment significantly decreased colony number and miR-21-3p level,but increased apoptotic rate and protein level of SMAD7 in a dose-dependent manner(P<0.05).Compared with cells treated with high-dose REM and transfected with miR-NC,those induced with high-dose REM and transfected with miR-21-3p mimics had significantly higher colony number and miR-21-3p level,but lower apoptotic rate and protein level of SMAD7(P<0.05).No significant differences in the colony number,apoptotic rate and expression levels of miR-21-3p and SMAD7 were detected between cells treated with high-dose REM and those treated with high-dose REM and transfected with miR-NC(P>0.05).Dual-luciferase reporter assay showed that the luciferase activity of WT-SMAD7 in the miR-21-3p mimics group was significantly lower than that in the miR-NC group(P<0.05).Conclusion REM affects the growth of TC cells by inhibiting the miR-21-3p/SMAD7 axis.
作者
王艳冰
毕啸
梁艳娇
王欣欣
刘海娟
廉皓
WANG Yanbing;BI Xiao;LIANG Yanjiao(Department of Anesthesiology,Chifeng Cancer Hospital,Inner Mongolia,Chifeng 024000,China)
出处
《河北医药》
2025年第12期1947-1951,共5页
Hebei Medical Journal
基金
内蒙古自治区自然科学基金(编号:2022MS08042)。
