Study on the Effect and Its Mechanism of Bromophenylcurcumin on Apoptosis ，Migration and Invasion of Cho- langiocarcinoma Cells

ZHAO Ji ’an; MA Yanrong; NIE Wenjia; DONG Liang; LIU Wencong; GU Jingfeng

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Study on the Effect and Its Mechanism of Bromophenylcurcumin on Apoptosis ，Migration and Invasion of Cho- langiocarcinoma Cells

更新时间：2022-06-21

- Study on the Effect and Its Mechanism of Bromophenylcurcumin on Apoptosis ，Migration and Invasion of Cho- langiocarcinoma Cells
- China Pharmacy Vol. 32, Issue 4, (2021)
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- Published：2021，
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ZHAO Ji ’an, MA Yanrong, NIE Wenjia, et al. Study on the Effect and Its Mechanism of Bromophenylcurcumin on Apoptosis ，Migration and Invasion of Cho- langiocarcinoma Cells. [J]. China Pharmacy 32(4).(2021)
DOI：

ZHAO Ji ’an, MA Yanrong, NIE Wenjia, et al. Study on the Effect and Its Mechanism of Bromophenylcurcumin on Apoptosis ，Migration and Invasion of Cho- langiocarcinoma Cells. [J]. China Pharmacy 32(4).(2021) DOI：

摘要

目的：研究溴苯基姜黄素（GL63）对人胆管癌RBE细胞凋亡、迁移和侵袭的影响，并基于Janus激酶（JAK）/信号转导及转录激活因子（STAT）信号通路探讨其作用机制。方法：采用MTT法检测不同浓度GL63[0（空白对照，下同）、1.25、2.5、5、10、20、40μmol/L]作用48h对RBE细胞增殖的影响，并计算其半数抑制浓度（IC50）；采用结晶紫染色法检测不同浓度GL63（0、5、10、20μmol/L）作用48h对细胞集落形成的影响；分别采用流式细胞术、Hoechst33342染色法、细胞划痕实验、Transwell小室侵袭实验检测不同浓度GL63（0、5、10、20μmol/L）作用24h对细胞周期分布、凋亡情况以及细胞迁移和侵袭能力的影响；采用Westernblotting法检测不同浓度GL63（0、5、10、20μmol/L）作用24h对细胞中JAK2/STAT3信号通路肿瘤相关蛋白表达的影响。结果：不同浓度GL63组（1.25～40μmol/L）RBE细胞的增殖抑制率均较空白对照组显著升高（P＜0.01），并呈剂量依赖趋势；其IC50为（8.46±1.30）μmol/L。与空白对照组比较，不同浓度GL63组（5、10、20μmol/L）RBE细胞的集落形成抑制率均显著降低（P＜0.01）；G0/G1期细胞占比均显著升高，S期细胞占比均显著降低（P＜0.01）；细胞凋亡率均显著升高（P＜0.01），且细胞核呈浓染致密的固缩形态并可见凋亡小体；细胞迁移愈合率均显著降低（P＜0.01），穿过基底膜的侵袭细胞数均显著减少（P＜0.01）；细胞中磷酸化JAK2、磷酸化STAT3、B淋巴细胞瘤-2（Bcl-2）、基质金属蛋白酶2（MMP-2）、MMP-9、前体胱天蛋白酶9（Pro-caspase-9）、Pro-caspase-3蛋白表达水平均显著降低，Bcl-2相关x蛋白（Bax）、细胞色素C、裂解Caspase-9（Cleaved-caspase-9）、Cleaved-caspase-3蛋白表达水平均显著升高（P＜0.01）。结论：GL63能通过抑制JAK2/STAT3信号通路来实现对人胆管癌RBE细胞增殖、迁移和侵袭的抑制并诱导其发生凋亡。

Abstract

OBJECTIVE：To study t he effects of bro mophenylcurcumin（GL63）on the apoptosis ，migration and invasion of human cholangiocarcinoma RBE cells ，and to investigate its mechanism based on JAK/STAT signaling pathway. METHODS ：MTT assay was used to detect the effects of different concentrations of GL 63 [0（blank control ，similarly hereinafter ），1.25，2.5，5，10， 20，40 μ mol/L] on the proliferation of RBE cells after 48 h treatment ；the IC 50 was calculated. The effects of different concentrations of GL 63（0，5，10，20 μmol/L）on colony formation were detected by crystal violet staining after 48 h treatment. Flow cytometry ，Hoechst 33342 staining，cell scratch test and Transwell chamber invasion test were used to detect the effects of different concentrations of GL 63（0，5，10，20 μmol/L）on cell cycle distribution ，apoptosis，migration and invasion ability after 24 h treatment. Western blotting assay was adopted to detect the effects of different concentration of GL 63（0，5，10，20 μmol/L） on the expression of JAK 2/STAT3 signal pathway associated proteins. RESULTS ：The proliferation inhibition rates of RBE cells in different concentrations of GL 63 groups（1.25-40 μmol/L）were significantly increase d，compared with blank control group （P＜ 0.01），and showed a dose-dependent trend ，with IC 50 of （8.46±1.30）μmol/L. Compared with blank control group， 85917439。E-mail：zhaoji-an-88@163.com inhibition rates of RBE cell colony formation were significantly decreased in different concentrations （5，10，20 μmol/L）of GL 63 groups（P＜0.01）. The percentage of RBE cells at G 0/G1 phase increased significantly ，while that at S phase decreased significantly （P＜0.01）. The apoptotic rate increased significantly（P＜0.01），and the nucleus showed dense pyknosis and apoptotic bodies. The rate of cell migration and healing was significantly decreased （P＜0.01），and the number of invasive cells through basement membrane was significantly decreased （P＜ 0.01）. The protein expression of p-JAK 2， p-STAT3， Bcl-2， MMP-2， MMP-9， Pro-caspase-9 and P ro-caspase-3 were down-regulated significantly while the expression of Bax ，Cyt-c，Cleaved-caspase-9 and Cleaved-caspase- 3 were up-regulated significantly（P＜0.01）. CONCLUSIONS ：GL63 may inhibit the proliferation ，migration and invasion of RBE cells and promote its apoptosis by inhibiting JAK 2/STAT3 signal pathway.

关键词

溴苯基姜黄素胆管癌RBE细胞Janus激酶2/信号转导及转录激活因子3信号通路迁移侵袭凋亡

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