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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
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| 篇名： | 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 |
| TITLE： | Study on regulation of ethanol extract of Turkish Galls on JAK 2/STAT3 signaling pathway in colorectal cancer cells |
| 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。 |
| ABSTRACT： | 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 |
| AUTHORS： | 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 |
| 阅读数： | 250 次 |
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