舒芬太尼调节cAMP/PKA/CREB信号通路对卵巢癌细胞增殖、凋亡和侵袭的影响  

Effects of Sufentanil on Proliferation,Apoptosis,and Invasion of Ovarian Cancer Cells by Regulating the cAMP/PKA/CREB Signaling Pathway

作　　者：甄磊[1] 张懿兰 王晓娜 焦颖 ZHEN Lei;ZHANG Yilan;WANG Xiaona;JIAO Ying(Department of Anesthesiology,the No.2 Hospital of Baoding,Baoding 071000,China;Department of Gynaecology,the No.2 Hospital of Baoding,Baoding 071000,China)

机构地区：[1]保定市第二医院麻醉科,保定071000 [2]保定市第二医院妇科,保定071000

出　　处：《中国细胞生物学学报》2024年第3期445-455,共11页Chinese Journal of Cell Biology

基　　金：保定市科技计划(批准号:2041ZF206)资助的课题。

摘　　要：该文旨在探究舒芬太尼(SFTN)调节环磷酸腺苷(cAMP)/蛋白激酶A(PKA)/环磷腺苷效应元件结合蛋白(CREB)信号通路对卵巢癌(OC)细胞增殖、凋亡和侵袭的影响。用浓度为2.5~160 ng/mL的舒芬太尼处理人OC细胞(SKOV-3),CCK-8法检测细胞活性,筛选最佳药物浓度。将SKOV-3细胞分为对照组(Control组),舒芬太尼低、中、高浓度组(SFTN-L组、SFTN-M组、SFTN-H组),舒芬太尼高浓度+PKA激活剂组(SFTN-H+8-溴-cAMP组),平板克隆法检测细胞增殖;流式细胞术检细胞凋亡;划痕实验检测细胞迁移;Transwell实验检测细胞侵袭;ELISA法检测环磷酸腺苷(cAMP)水平;Western blot法检测细胞核增殖抗原标记物(Ki67)、细胞周期蛋白D1(Cyclin D1)、半胱氨酸蛋白酶-3(Caspase-3)、B细胞淋巴瘤-2相关X蛋白(Bax)、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、蛋白激酶A(PKA)、磷酸化蛋白激酶A(p-PKA)、环磷腺苷效应元件结合蛋白(CREB)、磷酸化环磷腺苷效应元件结合蛋白(p-CREB)蛋白表达情况;裸鼠移植瘤实验检测舒芬太尼对OC移植瘤生长的影响。选择舒芬太尼浓度为20 ng/mL、40 ng/mL、80 ng/mL进行后续实验。与Control组比较,SFTN-L、SFTN-M、SFTN-H组的集落形成数、细胞划痕愈合率、细胞侵袭数量及Ki67、CyclinD1、MMP-2、MMP-9、cAMP、p-PKA/PKA、p-CREB/CREB表达水平降低,细胞凋亡率及Caspase-3、Bax表达水平显著升高,且呈浓度依赖性(P<0.05);与SFTN-H组相比,SFTN-H+8-溴-cAMP组的集落形成数、划痕愈合率、细胞侵袭数量及Ki67、Cyclin D1、MMP-2、MMP-9、cAMP、p-PKA/PKA、p-CREB/CREB表达水平升高,细胞凋亡率及Caspase-3、Bax表达水平显著降低(P<0.05)。移植瘤实验显示,SFTN组小鼠移植瘤比Control组生长缓慢,移植瘤质量、体积均减小,cAMP、p-PKA/PKA、p-CREB/CREB表达水平降低(P<0.05)。舒芬太尼通过抑制cAMP/PKA/CREB信号通路从而抑制OC细胞增殖和侵袭,促进细胞凋亡。This aim of this article was to investigate the effects of SFTN(sufentanil)modulating cAMP(cyclic adenosine monophosphate)/PKA(protein kinase B)/CREB(cAMP-response element binding protein)signaling pathway on the proliferation,apoptosis and invasion of OC(ovarian cancer)cells.Human SKOV-3(OC cells)were treated with sufentanil at concentrations ranging from 2.5 to 160 ng/mL,and cell activity was detected by the CCK-8 assay to screen for optimal drug concentration.SKOV-3 cells were divided into Control group,sufentanil low,medium,and high concentration groups(SFTN-L group,SFTN-M group,SFTN-H group),and sufentanil high concentration+PKA activator group(SFTN-H+8-bromo-cAMP group);and the plate cloning assay was used to detect cell proliferation;the apoptosis was detected by flow cytometry;the migration was detected by the scratch assay;the invasion was detected by the Transwell assay.ELISA method was applied to detect cAMP level.Western blot method was applied to detect Ki67(nuclear proliferative antigen markers),cyclin D1,Caspase-3,Bax(B-cell lymphoma as-sociated X-protein),MMP-2(matrix metalloproteinase-2),MMP-9(matrix metalloproteinase-9),PKA(protein kinase B),p-PKA(phosphorylated protein kinase B),CREB,p-CREB(phosphorylated-CREB)protein expression.Nude mice transplantation tumor assay to detect the effect of sufentanil on the growth of OC transplantation tumor.The ef-fect of sufentanil on the growth of OC transplanted tumors was detected in nude mice transplantation tumor assay.The concentrations of sufentanil were selected as 20 ng/mL,40 ng/mL,and 80 ng/mL for subsequent experiments.Com-pared with the Control group,the number of colony formation,cell scratch healing rate,number of cell invasion and the expression levels of Ki67,Cyclin D1,MMP-2,MMP-9,cAMP,p-PKA/PKA,and p-CREB/CREB were decreased in the SFTN-L,SFTN-M,and SFTN-H groups,and the apoptosis rate and the Caspase-3,Bax expression levels were significantly increased in a concentration-dependent manner(P<0.05);compared with the SFTN-H group,the num-ber of

关 键 词：卵巢癌 舒芬太尼 环磷酸腺苷(cAMP)/蛋白激酶A(PKA)/环磷腺苷效应元件结合蛋白(CREB)信号通路 增殖 凋亡 侵袭 

分 类 号：R737.31[医药卫生—肿瘤]