Effect of Drug-containing Serum of Clerodendrum bungei on Liver Cancer Cells Through PI3K/Akt Signaling Pathway

Qi HU; Ding-yao ZHU; Xiao-ning TAN; Ke-jian ZHU

doi:10.13422/j.cnki.syfjx.20191222

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Effect of Drug-containing Serum of Clerodendrum bungei on Liver Cancer Cells Through PI3K/Akt Signaling Pathway

Pharmacology | 更新时间：2021-02-09

- Effect of Drug-containing Serum of Clerodendrum bungei on Liver Cancer Cells Through PI3K/Akt Signaling Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 22, Pages: 28-33(2019)
- 作者机构：
  
  1.湖南中医药大学，长沙　 410208;
  2.湖南省中医药研究院　附属医院，长沙　 410006
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191222
  CLC： R22; R242; R2-031; R285.5
- Published：20 November 2019，
  
  Published Online：04 March 2019，
  
  Received：09 November 2018，
- 稿件说明：
扫描看全文
Qi HU, Ding-yao ZHU, Xiao-ning TAN, et al. Effect of Drug-containing Serum of Clerodendrum bungei on Liver Cancer Cells Through PI3K/Akt Signaling Pathway. [J]. Chinese Journal of Experimental Traditional Medical Formulae 25(22):28-33(2019)
DOI：

Qi HU, Ding-yao ZHU, Xiao-ning TAN, et al. Effect of Drug-containing Serum of Clerodendrum bungei on Liver Cancer Cells Through PI3K/Akt Signaling Pathway. [J]. Chinese Journal of Experimental Traditional Medical Formulae 25(22):28-33(2019) DOI： 10.13422/j.cnki.syfjx.20191222.

摘要

目的：

从磷脂酰肌醇3-激酶(PI3K)/丝氨酸苏氨酸蛋白激酶(Akt)信号通路角度探讨臭牡丹含药血清对肝癌MHCC97-H细胞的影响及可能的机制。

方法：

臭牡丹15%含药血清作用于MHCC97-H细胞，细胞增殖/毒性法(CCK-8)观察臭牡丹含药血清对细胞增殖的影响，以筛选最佳作用时间和浓度；Annexin V-FITC/碘化丙啶(PI)双染法检测细胞凋亡；蛋白免疫印迹法(Western blot)检测臭牡丹含药血清作用于细胞72 h后对PI3K/Akt信号通路中关键蛋白中第10号染色体缺失的磷酸酶及张力蛋白同源的基因(PTEN)，磷酸化Akt(p-Akt)和磷脂酰肌醇3-激酶(PI3K)蛋白表达的影响；实时荧光定量聚合酶链式反应(Real-time PCR)检测臭牡丹含药血清作用于细胞72 h后对核转录因子-

B(NF-

B)和肿瘤坏死因子-

(TNF-

)mRNA表达的影响。

结果：

CCK-8结果显示，与空白组比较，臭牡丹含药血清对肿瘤细胞的增殖抑制作用呈剂量和时间依赖性。其中高剂量组抑制作用最为明显，在24，48，72 h的最大抑制率分别达到28%，32%，43%；流式细胞术结果显示，与空白组比较，随着臭牡丹含药血清浓度的增加，细胞的生长均受不同程度抑制，细胞的凋亡比例亦相应增加，72 h干预组抑制作用显著(

0.01)，臭牡丹含药血清组各时间段最大凋亡率达19.48%，19.72%(

0.05)；Western blot结果显示，与空白组比较，臭牡丹含药血清干预下各组PTEN表达量均升高(

0.05)，PI3K，p-Akt蛋白表达均减少(

0.05)；Real-time PCR结果显示，与空白组比较，臭牡丹含药血清可以下调NF-

B mRNA表达，上调TNF-

mRNA的表达(

0.05)。

结论：

臭牡丹含药血清可以有效抑制MHCC97-H肝癌细胞增殖并促进其凋亡，可能与PI3K/Akt信号通路并影响其关键因子有关。

Abstract

Objective:

To investigate the effect of

Clerodendrum bungei

-containing serum on liver cancer MHCC97-H cells and its possible mechanism from the perspective of phosphatidylinositol 3-kinase(PI3K)/protein kinase (Akt) signaling pathway.

Method:

The medicinal serum of 15%

bungei

was used to treat MHCC97-H cells. The effect of serum containing

bungei

on cell proliferation was observed by cell counting kit-8(CCK-8) method

in order to select the best time and concentration. The apoptosis was detected by Annexin V-FITC/PI double staining method. Western blot was used to detect the posphatase and tensin homologous gene deleted from chromosome 10 in key proteins (PTEN)

phosphoprotein kinase B (p-Akt) and phosphatidylinositol 3-kinase (PI3K)-related protein expression of PI3K/Akt signaling pathway. Real-time PCR was used to detect

bungei

-containing serum on cells for 72 h after activation of nuclear factor-activated B cell kappa light chain(NF-

B) and tumor necrosis factor-

(TNF-

) mRNA expression.

Result:

The results of CCK-8 showed an inhibitory effect of the

bungei

-containing serum on the proliferation of tumor cells in a dose and time-dependent manner. Among them

the high-dose group had the most obvious inhibitory effect

and the maximum inhibition rates at 24

72 h were 28%

32%

and 43%

respectively. The results of flow cytometry showed that with the increase of drug-containing serum concentration

the cell growth was observed. The inhibition rate of cells was increased to different degrees

and the inhibition effect was significantly increased in the 72 h intervention group (

0.01). The maximum apoptosis rate of the

bungei

-containing serum group was 19.48% and 19.72%

compared with the blank group. The difference was significant (

0.05). The results of Western blot showed that the expression of PTEN in different groups increased with different concentrations of

bungei

-containing serum (

0.05)

and the expressions of PI3K and p-Akt decreased (

0.05). Real-time PCR results showed that the

bungei

-containing serum could down-regulate the expression of NF-

B and up-regulate the expression of TNF-

mRNA (

0.05).

Conclusion:

The medicinal serum of

bungei

can effectively inhibit the proliferation of MHCC97-H hepatoma cells and promote its apoptosis

which may be related to the PI3K/Akt signaling pathway and its key factors.

关键词

臭牡丹肝癌细胞MHCC97-H磷脂酰肌醇3-激酶(PI3K)/丝氨酸苏氨酸蛋白激酶(Akt)含药血清

Keywords

Clerodendron bungeiliver cancer cell MHCC97-Hphosphatidylinositol 3-kinase(PI3K)/protein kinase (Akt) signaling pathwaydrug-containing serum

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