Effect of Oxymatrine in Inhibiting Proliferation and Migration of HT-29 Cells Induced by Insulin

Di PAN; Wen ZHANG; Rong ZHANG; Shi-quan GAN; Yan CHEN; Nen-ling ZHANG; Yi-ni XU; Xiang-chun SHEN

doi:10.13422/j.cnki.syfjx.20192022

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Effect of Oxymatrine in Inhibiting Proliferation and Migration of HT-29 Cells Induced by Insulin

Pharmacology | 更新时间：2021-02-09

- Effect of Oxymatrine in Inhibiting Proliferation and Migration of HT-29 Cells Induced by Insulin
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 24, Pages: 36-42(2019)
- 作者机构：
  
  1.贵州省天然药物资源高效利用工程中心，贵阳　 550025；
  2.贵州医科大学　天然药物资源优效利用重点实验室，贵阳　 550025
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20192022
  CLC： R22; R242; R2-031; R285.5
- Received：27 January 2019，
  
  Published Online：24 June 2019，
  
  Published：20 December 2019
- 稿件说明：
移动端阅览
Di PAN, Wen ZHANG, Rong ZHANG, et al. Effect of Oxymatrine in Inhibiting Proliferation and Migration of HT-29 Cells Induced by Insulin[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(24): 36-42.
DOI：

Di PAN, Wen ZHANG, Rong ZHANG, et al. Effect of Oxymatrine in Inhibiting Proliferation and Migration of HT-29 Cells Induced by Insulin[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(24): 36-42. DOI： 10.13422/j.cnki.syfjx.20192022.

摘要

目的：

利用胰岛素培养人结肠癌细胞HT-29，体外模拟糖尿病患者体循环中高胰岛素环境，研究氧化苦参碱(OMT)对胰岛素诱导的人结肠癌HT-29细胞增殖和迁移的影响。

方法：

采用噻唑蓝(MTT)比色法和平板克隆实验检测OMT(2

8 mmol·L

－1

)对胰岛素诱导HT-29的增殖作用，倒置显微镜观察该模型下细胞形态的变化；通过划痕修复实验评价OMT对胰岛素诱导的HT-29迁移能力，流式细胞术Annexin V/碘化丙啶(PI)双染实验检测该模型中HT-29细胞周期与凋亡的变化；蛋白免疫印迹法(Western blot)检测细胞周期相关蛋白的表达以及细胞迁移相关蛋白的表达。

结果：

MTT比色法、平板克隆及划痕修复实验均显示，与空白组比较，胰岛素能促进人结肠癌HT-29细胞的增殖和迁移(

0.05)；选择2

8 mmol·L

－1

OMT处理细胞24 h，与胰岛素组比较，OMT 4

8 mmol·L

－1

明显抑制其增殖作用(

0.05)，可诱导HT-29细胞发生G

期阻滞(

0.05)，8 mmol·L

－1

OMT下调细胞周期蛋白D

(CyclinD

)和周期蛋白依赖性激酶4(CDK4)表达(

0.05)，上调周期负调节蛋白p27水平(

0.05)；此外，划痕修复实验结果显示，与胰岛素组比较，OMT各浓度组均能抑制胰岛素诱导的细胞迁移作用(

0.05)，其机制可能与增加黏附蛋白(E-cadherin)(

0.05)和降低波形蛋白(Vimentin)(

0.05)相关。

结论：

OMT能抑制胰岛素诱导的人结肠癌HT-29细胞增殖及迁移作用，其作用机制可能与调控细胞周期及迁移相关蛋白有关。

Abstract

Objective:

To investigate the effect of oxymatrine (OMT) on the proliferation and migration of human colon cancer cell line HT-29 under Type Ⅱ diabetes environment by co-culturing HT-29 with insulin to simulate hyperinsulinemia.

Method:

The effect of OMT (2

8 mmol·L

-1

) on insulin-induced proliferation of HT-29 was detected by methyl thiazolyl tetrazolium (MTT) assay and cloning assay. The morphology change and cell migration were evaluated under microscope and by wound healing assay. The Annexin V/propidium iodide(PI) assay was used to detect the change of insulin-induced HT-29 cell cycle and apoptosis. Western blot was performed to validate the expression of cell cycle-related protein and cell migration protein.

Result:

Insulin significantly increased growth of HT-29 (

0.05). Compared with insulin group

OMT with 2

8 mmol·L

-1

showed a significant inhibitory effect in this model (

0.05). In addition

OMT blocked HT-29 cell cycle in G

phase (

0.05)

and showed a slight apoptotic effect. Western blot showed that the down-regulation of Cyclin D

CDK4 and the up-regulation of p27 by OMT might involve the growth inhibition mechanism. Furthermore

OMT reduced the migration of insulin-induced HT-29 according to wound healing assay(

0.05). Decreased Vimentin (

0.05)and increased E-cadherin(

0.05)might be correlated with the migration restrain.

Conclusion:

OMT can inhibit the proliferation and migration of insulin-induced HT-29 cells. The changes of cell cycle and migration related proteins may be correlated with the mechanism.

关键词

Keywords

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