丹参酮ⅡA抑制STAT3通路调节食管癌EC109细胞的周期与凋亡  

TanshinoneⅡA Regulating Cycle and Apoptosis of Esophageal Cancer EC109 Cells by Inhibiting STAT3 Pathway

作　　者：陈星[1] 周哲旭 尚艺婉 刘洋[1] 刘娅茹 胡啸博 樊李妍 陈玉龙[1] CHEN Xing;ZHOU Zhexu;SHANG Yiwan;LIU Yang;LIU Yaru;HU Xiaobo;FAN Liyan;CHEN Yulong(Henan University of Chinese Medicine/Henan Provincial Key Laboratory of Chinese Medicine Signal Transduction,Zhengzhou Henan China 450046)

机构地区：[1]河南中医药大学/河南省中医方证信号传导重点实验室,河南郑州450046

出　　处：《中医学报》2023年第11期2414-2424,共11页Acta Chinese Medicine

基　　金：国家自然科学基金面上项目(82074313)。

摘　　要：目的:观察丹参酮ⅡA能否通过抑制信号传导及转录激活蛋白3(signal transducer and activator of transcription 3,STAT3)信号通路影响食管癌EC109细胞的周期与凋亡。方法:对数生长期的EC109细胞分为空白组、不同浓度丹参酮ⅡA组、不同浓度stattic组及丹参酮ⅡA和stattic联合用药组,MTT法检测丹参酮ⅡA、stattic及联合用药对EC109细胞增殖的影响;流式细胞术检测细胞周期及凋亡;从70只4周龄SPF级BALB/c雌鼠中随机抽取10只作为空白组,其余小鼠右胁部皮下注射2×106个Ec109细胞建立食管癌模型。将造模成功的小鼠随机分为模型组、丹参酮ⅡA低浓度组(20 mg·kg^(-1))、丹参酮ⅡA高浓度组(40 mg·kg^(-1))、stattic组(25 mg·kg^(-1))、stattic(25 mg·kg^(-1))+丹参酮ⅡA高浓度(40 mg·kg^(-1))组、顺铂组(1 mg·kg^(-1)),腹腔注射给药,每2天进行1次,连续35 d。给药期间每7天测量一次瘤体大小与小鼠体质量,考察药物对小鼠肿瘤生长的抑制作用;ELISA法检测移植瘤裸鼠血清中白细胞介素-6(interleukin-6,IL-6)的水平;Western Blot、RT-PCR检测食管癌EC109细胞与小鼠肿瘤组织细胞周期、凋亡相关蛋白及mRNA的表达水平。结果:细胞实验发现,与空白组比较,丹参酮ⅡA、stattic均可明显抑制EC109细胞增殖(P<0.01);丹参酮ⅡA能显著促进细胞进入G2期,促进其凋亡(P<0.05)。Western Blot及RT-PCR结果显示,丹参酮ⅡA能够抑制STAT3、p-STAT3、c-Myc、CyclinB1蛋白及mRNA的表达,促进凋亡因子Caspase-3、Caspase-9蛋白及mRNA的表达(P<0.05)。体内动物实验结果表明,与模型组比较,丹参酮ⅡA、stattic均能明显抑制裸鼠移植瘤生长(P<0.01),降低血清IL-6水平(P<0.05),抑制STAT3、p-STAT3、c-Myc蛋白及mRNA的表达(P<0.05),促进凋亡因子Caspase-3、Caspase-9蛋白及mRNA的表达(P<0.05);与stattic比较,联合用药组Caspase-3 mRNA、Caspase-9蛋白及mRNA的表达水平显著降低(P<0.05)。结论:丹参酮ⅡA能够诱导食管癌�Objective:To observe whether tanshinoneⅡA can affect the cycle and apoptosis of esophageal cancer EC109 cells by inhibiting signal transduction and STAT3 signaling pathway.Methods:EC109 cells in logarithmic growth period were divided into blank group,tanshinoneⅡA group with different concentrations,stattic group with different concentrations and tanshinoneⅡA and stattic combined group.The effects of tanshinoneⅡA,stattic and their combined drugs on the proliferation of EC109 cells were detected by MTT method.Cell cycle and apoptosis were detected by flow cytometry.Ten of 70 four-week-old SPF BALB/c female mice were randomly selected as blank group,and the other mice were subcutaneously injected with 2×106 Ec109 cells in the right hypochondrium to establish esophageal cancer model.The successful mice were randomly divided into model group,tanshinoneⅡA low concentration group(20 mg·kg^(-1)),tanshinoneⅡA high concentration group(40 mg·kg^(-1)),stattic group(25 mg·kg^(-1)),stattic(25 mg·kg^(-1))+tanshinoneⅡA high concentration group(40 mg·kg^(-1))and cisplatin group(1 mg·kg^(-1)).The tumor size and weight of mice were measured every 7 days during the administration period to investigate the inhibitory effect of drugs on tumor growth in mice.The level of interleukin-6(IL-6)in serum of nude mice with transplanted tumor was detected by ELISA.WesternBlot and RT-PCR were used to detect the expression levels of cell cycle,apoptosis-related proteins and mRNA in EC109 cells and tumor tissues of mice.Results:Compared with the blank group,tanshinoneⅡA and stattic could significantly inhibit the proliferation of EC109 cells(P<0.01).TanshinoneⅡA can significantly promote cells to enter G2 phase and promote their apoptosis(P<0.05).WesternBlot and RT-PCR results suggusted that tanshinoneⅡA could inhibit the expression of STAT3,p-STAT3,c-Myc,CyclinB1 protein and mRNA,and promote the expression of apoptosis factor Caspase-3,Caspase-9 protein and mRNA(P<0.05).The results of animal experiments in vivo showed

关 键 词：丹参酮ⅡA 食管癌 STAT3信号通路 细胞周期 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]