没食子乙醇提取物对结肠癌细胞JAK2/STAT3信号通路的调控作用研究 点击下载
论文标题: 没食子乙醇提取物对结肠癌细胞JAK2/STAT3信号通路的调控作用研究
英文标题:
中文摘要: 目的 基于蛋白酪氨酸激酶2(JAK2)/信号传导及转录活化因子3(STAT3)信号通路研究没食子乙醇提取物对结肠癌细胞HCT-116、Caco-2增殖、迁移的调控机制。方法采用CCK-8法检测0.05、0.1、0.2、0.3、0.4、0.5mg/mL没食子乙醇提取物作用12、24、48、72h后对HCT-116、Caco-2细胞增殖的影响。以0.1、0.3、0.5mg/mL没食子乙醇提取物作用于HCT-116、Caco-2细胞24h后,采用划痕实验测定细胞的迁移情况,采用荧光探针法检测细胞内活性氧(ROS)水平,采用酶联免疫吸附法检测细胞上清液中白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)水平,采用Westernblot法检测细胞中JAK2、STAT3磷酸化水平和B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)的表达水平。结果与空白对照比较,0.05、0.1、0.2、0.3、0.4、0.5mg/mL没食子乙醇提取物作用12、24、48、72h后均可显著抑制细胞增殖(P<0.05)。0.1、0.3、0.5mg/mL没食子乙醇提物作用24h后,2种细胞的划痕愈合率和细胞上清液中IL-6、TNF-α水平,以及细胞中JAK2、STAT3磷酸化水平和Bcl-2蛋白表达水平均显著降低(P<0.05);细胞内ROS水平、Bax蛋白表达水平均显著升高(P<0.05)。结论没食子乙醇提取物可抑制结肠癌细胞HCT-116、Caco-2的增殖和迁移,其机制可能与增加细胞内ROS的积累,下调肿瘤微环境中炎症因子IL-6、TNF-α的表达和JAK2/STAT3信号通路中JAK2、STAT3的磷酸化水平,进而下调Bcl-2蛋白表达、上调Bax蛋白表达有关。
英文摘要: OBJECTIVE To study the regulatory mechanism of ethanol extract of Turkish Galls on proliferation and migration of colorectal cancer cells HCT- 116 and Caco- 2 based on janus kinase 2(JAK2)/signal transducer and activator of transcription 3 (STAT3)signaling pathway. METHODS CCK-8 method was used to detect the effects of 0.05,0.1,0.2,0.3,0.4 and 0.5 mg/mL ethanol extract of Turkish Galls on the proliferation of HCT- 116 and Caco- 2 cells after treated for 12,24,48 and 72 h. After treated with 0.1,0.3,0.5 mg/mL ethanol extract of Turkish Galls for 24 h,the migrations of HCT- 116 and Caco- 2 cells were detected by scratch test ;the level of reactive oxygen species (ROS)was detected by fluorescent probe method. The levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in cell supernatant were detected by ELISA . The phosphorylations of JAK2 and STAT 3 as well as the expressions of B-cell lymphoma- 2(Bcl-2)and Bcl- 2 associated protein X (Bax)were detected by Western blot assay. RESULTS Compared with blank control ,0.05,0.1,0.2,0.3,0.4 and 0.5 mg/mL ethanol extract of Turkish Galls could significantly inhibit cell proliferation after treated for 12,24,48,72 h(P<0.05). After treated with 0.1,0.3 and 0.5 mg/mL ethanol extract of Turkish Galls for 24 h,the scratch healing rate of 2 kinds of cells ,the levels of IL- 6 and TNF-α in the cell supernatant ,the phosphorylation of JAK 2 and STAT 3 as well as the expression of Bcl- 2 protein were all significantly decreased (P<0.05);the level of ROS and protein expression of Bax were increased significantly (P<0.05). CONCLUSIONS The ethanol extract of Turkish Galls can inhibit the proliferation and migration of HCT- 116 and Caco- 2 cells. The mechanism may be related with down-regulation of protein expression of Bcl- 2 and up-regulation of protein expression of Bax by increasing the accumulation of intracellular ROS ,down-regulating the expressions of inflammatory factors IL- 6 and TNF-α and the phosphorylation of JAK2 and STAT 3 in JAK 2/STAT3 signaling pathway.
期刊: 2022年第33卷第03期
作者: 阿丽亚·依拉木,阿布都艾则孜·艾尔肯,闫波,艾尔菲丁·阿尼娃尔,木巴拉克·伊明江
英文作者: Aliya·Elham,Abdulaziz·Arken,YAN Bo ,Arfidin·Anwar,Mubarak·Iminjan
关键字: 没食子乙醇提取物;结肠癌细胞;炎症因子;活性氧;蛋白酪氨酸激酶2/信号传导及转录活化因子3信号通路
KEYWORDS: ethanol extract of Turkish Galls ;colorectal
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