Mechanism of Biejiajian Wan Against EMT of Hepatocellular Carcinoma Cells Through NF-<italic style="font-style: italic">κ</italic>B Signaling Pathway

Xiao-dan ZHONG; Bin WEN; Hai-tao SUN; Jia-ling SUN; Xue-mei YANG; Wei-cong CHEN; Wen-ting ZHAO; Chun-yu HE; Yang LIU; Tong LI; Song-qi HE

doi:10.13422/j.cnki.syfjx.20212421

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Mechanism of Biejiajian Wan Against EMT of Hepatocellular Carcinoma Cells Through NF-κB Signaling Pathway

更新时间：2022-01-04

- Mechanism of Biejiajian Wan Against EMT of Hepatocellular Carcinoma Cells Through NF-κB Signaling Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 28, Issue 1, Pages: 24-32(2022)
- 作者机构：
  
  1.南方医科大学中医药学院，广州 510515
  2.解放军南部战区空军医院，广州 510602
  3.南方医科大学中西医结合医院，广州 510315
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20212421
  CLC： R22;R242;R2-031;R285.5
- Received：26 July 2021，
  
  Published Online：22 October 2021，
  
  Published：05 January 2022
- 稿件说明：
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钟晓丹,文彬,孙海涛等.鳖甲煎丸通过NF-κB信号通路抑制肝癌细胞上皮间质转化的作用机制[J].中国实验方剂学杂志,2022,28(01):24-32.

ZHONG Xiao-dan,WEN Bin,SUN Hai-tao,et al.Mechanism of Biejiajian Wan Against EMT of Hepatocellular Carcinoma Cells Through NF-κB Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(01):24-32.
钟晓丹,文彬,孙海涛等.鳖甲煎丸通过NF-κB信号通路抑制肝癌细胞上皮间质转化的作用机制[J].中国实验方剂学杂志,2022,28(01):24-32. DOI： 10.13422/j.cnki.syfjx.20212421.

ZHONG Xiao-dan,WEN Bin,SUN Hai-tao,et al.Mechanism of Biejiajian Wan Against EMT of Hepatocellular Carcinoma Cells Through NF-κB Signaling Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(01):24-32. DOI： 10.13422/j.cnki.syfjx.20212421.

摘要

目的

研究鳖甲煎丸对转化生长因子-

（TGF-

）诱导HepG2细胞上皮间质转化（EMT）的影响，探讨其抑制肝癌细胞EMT的作用机制。

方法

以HepG2细胞为研究对象，将细胞随机分为空白组，模型组（10 μg·L

-1

TGF-

），鳖甲煎丸低（10 μg·L

-1

TGF-

+0.55 g·kg

-1

鳖甲煎丸）、中（10 μg·L

-1

TGF-

+1.1 g·kg

-1

鳖甲煎丸）、高（10 μg·L

-1

TGF-

+2.2 g·kg

-1

鳖甲煎丸）组，索拉非尼组（10 μg·L

-1

TGF-

+0.03 g·kg

-1

索拉非尼）。用10 μg·L

-1

TGF-

诱导HepG2细胞发生EMT，构建EMT模型。分别给予相应的含药血清处理后，采用细胞增殖与活性检测（CCK-8）法检测细胞增殖情况，分别采用细胞划痕实验，transwell迁移实验检测细胞迁移情况，采用免疫荧光、蛋白免疫印迹法分别检测EMT，核转录因子-

B（NF-

B）信号通路相关蛋白的表达情况。

结果

与空白组比较，TGF-

刺激4 d后，细胞呈梭形改变，细胞之间变得松散，间隙增宽，伸出触角，同时E-钙黏素（E-cadherin）蛋白表达降低（

0.05），N-钙黏素（N-cadherin），波形蛋白（vimentin）蛋白表达升高（

0.05），说明TGF-

刺激4 d成功构建了HepG2细胞的EMT模型组。与模型组比较，用相应含药血清处理48 h，各用药组均能抑制已发生EMT作用的HepG2细胞增殖，其中以鳖甲煎丸低、高剂量组最为显著（

0.01）。与模型组比较，鳖甲煎丸中剂量组E-cadherin蛋白表达升高（

0.05），磷酸化（p）-p65，N-cadherin，vimentin蛋白表达降低（

0.05）。与空白组比较，细胞划痕实验和transwell迁移实验表明TGF-

增强了HepG2细胞的迁移能力（

0.05，

0.01）；与模型组比较，鳖甲煎丸低、中、高剂量组和索拉非尼组抑制HepG2细胞迁移能力（

0.05，

0.01）。与空白组比较，模型组促进p65，Snail入核表达；与模型组比较，鳖甲煎丸低、中、高剂量组和索拉非尼组抑制p65和Snail入核表达。

结论

鳖甲煎丸可能通过抑制NF-

B信号通路来抑制TGF-

诱导HepG2细胞的EMT进程、增殖和迁移，从而发挥抗肝癌的作用。

Abstract

Objective

To investigate the effect of Biejiajian Wan （BJJW） on transforming growth factor-

（TGF-

）-induced epithelial-mesenchymal transition （EMT） of HepG2 cells， and explore its mechanism against EMT of hepatocellular carcinoma cells.

Method

HepG2 cells were randomly divided into a blank group， a TGF-

model group （10 μg·L

-1

TGF-

）， a low-dose BJJW group （10 μg·L

-1

TGF-

+0.55 g·kg

-1

BJJW）， a medium-dose BJJW group （10 μg·L

-1

TGF-

+1.1 g·kg

-1

BJJW）， a high-dose BJJW group （10 μg·L

-1

TGF-

+2.2 g·kg

-1

BJJW）， and a sorafenib group （10 μg·L

-1

TGF-

+0.03 g·kg

-1

sorafenib）. The EMT model was induced by 10 μg·L

-1

TGF-

HepG2 cells. After treatment with corresponding medicated serum， cell counting kit -8 （CCK-8） assay was used to detect cell proliferation. Cell migration ability was detected by the Transwell assay and wound healing assay. The protein expression related to EMT and nuclear factor-kappa B （NF-

B） signaling pathway was detected by cell immunofluorescence assay and Western blot.

Result

Compared with the blank group 4 days later， the TGF-

model group showed fusiform and loose cells with widened gap and antennae reaching out， decreased protein expression of E-cadherin （

0.05）， and increased protein expression of N-cadherin and vimentin （

0.05）， which indicated that the EMT model was properly induced in HepG2 cells by TGF-

stimulation for 4 days. After 48 hours of treatment with the corresponding medicated serum， each medication group showed inhibited proliferation of HepG2 cells that had undergone EMT， especially the low- and high-dose BJJW groups （

0.01）， and the medium-dose BJJW group showed increased E-cadherin protein expression （

0.05） and decreased p-p65， N-cadherin， and vimentin protein expression （

0.05）， as compared with the TGF-

model group. As revealed by the transwell assay and wound healing assay， TGF-

enhanced the migration ability of HepG2 cells （

0.05，

0.01） compared with the results in the blank group， compared with the TGF-

model group， the medication groups showed inhibited migration ability of HepG2 cells （

0.05，

0.01）. Compared with the blank group， the TGF-

model group promoted the expression of p65 and Snail into the nucleus. Compared with the TGF-

model group， the medication groups inhibited the expression of p65 and Snail into the nucleus.

Conclusion

BJJW may inhibit the EMT， proliferation， and migration of HepG2 cells induced by TGF-

by suppressing the NF-

B signaling pathway to exert an anti-hepatocellular carcinoma effect.

关键词

Keywords

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Mechanism of Biejiajian Wan Against EMT of Hepatocellular Carcinoma Cells Through NF-κB Signaling Pathway

DOI：10.13422/j.cnki.syfjx.20212421

摘要

Abstract

关键词

Keywords

references