舒芬太尼诱导宫颈癌细胞自噬和凋亡并抑制癌细胞恶性增殖  

Sufentanil Induces Autophagy and Apoptosis of Cervical Cancer Cells and Inhibits Their Proliferation

作　　者：张贤雨[1] 马欢[1] 宋凤丽 刘晓玉 李植燕 原娜[1] 郝晓慧 张志林[1] ZHANG Xianyu;MA Huan;SONG Fengli;LIU Xiaoyu;LI Zhiyan;YUAN Na;HAO Xiaohui;ZHANG Zhilin(Department of Radiotherapy,the First Affiliated Hospital of Hebei North University,Zhangjiakou,Hebei,075000,China;Department of Radiotherapy,the Third Affiliated Hospital of Beijing University of Chinese Medicine,Beijing,100029,China)

机构地区：[1]河北北方学院附属第一医院放疗科,河北省张家口市075000 [2]北京中医药大学第三附属医院放疗科,北京市100029

出　　处：《医学分子生物学杂志》2022年第3期219-224,共6页Journal of Medical Molecular Biology

基　　金：张家口市2021年市级科技计划自筹经费项目(No.2121192H)。

摘　　要：目的探讨舒芬太尼对宫颈癌细胞SiHa自噬、凋亡及细胞增殖的影响。方法采用CCK8法检测舒芬太尼梯度浓度(3.125、6.25、12.5、25、50、100、200、400、800 nmol/L)作用下宫颈癌细胞SiHa细胞活力,选取4个舒芬太尼作用浓度(0、25、50、100 nmol/L)处理SiHa细胞24 h,采用流式细胞法、克隆形成实验检测SiHa细胞凋亡及增殖情况,免疫荧光法检测各组细胞LC3水平,采用Western印迹检测自噬相关蛋白LC3Ⅱ/LC3Ⅰ、Beclin1、ATG7与增殖、凋亡相关蛋白P21、Survivin水平,采用RT-PCR检测细胞增殖相关基因Ki67、PCNA表达水平。结果随着舒芬太尼处理浓度的增加,SiHa细胞活力逐渐降低。50、100 nmol/L舒芬太尼处理下SiHa细胞的LC3Ⅱ/LC3Ⅰ、Beclin1、ATG7水平显著高于0 nmol/L舒芬太尼处理,免疫荧光检测结果显示50、100 nmol/L舒芬太尼处理下LC3荧光信号高于0 nmol/L舒芬太尼处理,流式细胞仪检测结果显示50、100 nmol/L舒芬太尼处理下SiHa细胞凋亡高于0 nmol/L舒芬太尼处理。克隆形成实验结果显示50、100 nmol/L舒芬太尼处理下SiHa细胞克隆形成率降低,Survivin水平与Ki67、PCNA表达水平降低,而P21水平升高。结论舒芬太尼对宫颈癌细胞自噬、凋亡有促进作用,对细胞增殖有抑制作用,可能与调节细胞自噬、凋亡、增殖相关蛋白或基因水平有关。Objective To explore the effects of sufentanil on autophagy,apoptosis and proliferation of cervical cancer cells SiHa.Methods SiHa cells were treated with sufentanil(3.125,6.25,12.5,25,50,100,200,400,800 nmol/L).Cell viability was measured by CCK8.Four concentrations of sufentanil(0,25,50,100 nmol/L)were then selected to treat Si-Ha cells for 24 h for the following experiments.The apoptosis and proliferation of SiHa cells were measured by flow cytometry and colony formation assay.The level of LC3 in each group was measured by immunofluorescence method.The levels of autophagy-related proteins(LC3Ⅱ/LC3Ⅰ,Beclin1,ATG7),proliferation-and apoptosis-related proteins(P21,Survivin)were detected by Western blotting.The expression levels of cell proliferation-related genes(Ki67,PCNA)were detected by RT-PCR.Results The viability of SiHa cells decreased with the increased concentration of sufentanil and the effect was in a dose-dependent manner.The expression levels of LC3Ⅱ/LC3Ⅰ,Beclin1 and ATG7 in SiHa cells treated with 50 nmol/L and 100 nmol/L sufentanil were significantly higher than those treated with 0 nmol/L sufentanil.The immunofluorescence method showed that the fluorescence signal of LC3 in SiHa cells treated with 50 and 100 nmol/L sufentanil was higher than that in cells treated with 0 nmol/L sufentanil.Flow cytometry showed that the apoptosis rate of SiHa cells treated with 50 and 100 nmol/L sufentanil was higher than that of cells treated with 0 nmol/L sufentanil.Clone formation assay showed that the clonal formation rate of SiHa cells was decreased after 50 and 100 nmol/L sufentanil treatment.The expression levels of Survivin,Ki67 and PCNA were decreased,and the expression level of P21 was increased.Conclusion Sufentanil can promote autophagy and apoptosis of cervical cancer cells,and inhibit their proliferation,which may be through the regulation of autophagy-,apoptosis-and proliferation-related proteins or genes.

关 键 词：宫颈癌 舒芬太尼 细胞自噬 细胞凋亡 细胞增殖 

分 类 号：R737.3[医药卫生—肿瘤]