Effect of Modified Si Junzitang Drug Serum on Expression of Apoptosis-related Molecules of Gastric Cancer Cell SGC-7901

Shan-shan NIE; Xun LI; Yu-hang ZHAO; Dong-sheng WANG

doi:10.13422/j.cnki.syfjx.20190824

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Effect of Modified Si Junzitang Drug Serum on Expression of Apoptosis-related Molecules of Gastric Cancer Cell SGC-7901

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- Effect of Modified Si Junzitang Drug Serum on Expression of Apoptosis-related Molecules of Gastric Cancer Cell SGC-7901
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 9, Pages: 25-30(2019)
- 作者机构：
  
  中南大学　湘雅医院，长沙　 410008
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20190824
  CLC： R22; R242; R2-031; R285.5
- Received：23 August 2018，
  
  Published Online：04 January 2019，
  
  Published：05 May 2019
- 稿件说明：
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Shan-shan NIE, Xun LI, Yu-hang ZHAO, et al. Effect of Modified Si Junzitang Drug Serum on Expression of Apoptosis-related Molecules of Gastric Cancer Cell SGC-7901[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(9): 25-30.
DOI：

Shan-shan NIE, Xun LI, Yu-hang ZHAO, et al. Effect of Modified Si Junzitang Drug Serum on Expression of Apoptosis-related Molecules of Gastric Cancer Cell SGC-7901[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(9): 25-30. DOI： 10.13422/j.cnki.syfjx.20190824.

摘要

目的：

研究加味四君子汤含药血清对胃癌细胞SGC-7901凋亡相关因子表达的影响，进一步探讨其具体抗肿瘤作用机制。

方法：

40只SD大鼠随机分为加味四君子汤低、中、高剂量组(0.213，0.426，0.853 g·kg

－1

)，空白组，每组10只，空白组给予等体积的生理盐水灌胃，各组连续灌胃10 d，末次给药1.5 h后，水合氯醛腹腔麻醉，心脏采血，分离血清，56 ℃灭菌，0.22 μm过滤，制备加味四君子汤高、中、低剂量组含药血清，各剂量组含药血清孵育胃癌细胞SGC-7901，运用流式细胞仪检测细胞早期和晚期凋亡情况，运用实时荧光定量聚合酶链式反应(Real-time PCR)技术检测肿瘤抑制基因p53，c-核蛋白类基因(c-Myc)，半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)，凋亡相关基因B细胞淋巴瘤-2(Bcl-2) mRNA的表达，运用细胞免疫荧光技术检测p53，c-Myc

Caspase-3，Bcl-2蛋白的相对表达量情况。

结果：

与空白组比较，加味四君子汤含药血清高剂量组细胞凋亡比率显著升高(

0.01)，且可使胃癌细胞SGC-7901早期＋晚期凋亡率达到22.58%(

0.01)。Real-time PCR结果显示加味四君子汤中剂量组能显著促进Caspase-3 mRNA表达(

0.01)，加味四君子汤高剂量组能显著上调p53及c-Myc mRNA表达量(

0.01)，加味四君子汤高剂量组显著抑制Bcl-2 mRNA表达(

0.01)。免疫荧光结果显示加味加味四君子汤高剂量组能使c-Myc

Caspase-3，p53蛋白表达水平显著升高(

0.01)，而Bcl-2蛋白表达水平显著降低(

0.05)。

结论：

加味四君子汤含药血清能抑制抗凋亡蛋白Bcl-2的表达，促进凋亡相关分子p53，c-Myc

Caspase-3的表达，发挥抗肿瘤作用。

Abstract

Objective:

To explore the effect of modified Si Junzitang (MSJZT) drug serum on the expression of apoptosis-related molecules of gastric cancer cell SGC-7901 and further its anti-tumor mechanism.

Method:

A total of 40 SD rats were randomly divided into four groups: low-dose

middle-dose

high-dose MSJZT (0.213

0.426

0.853 g·kg

-1

) groups and normal group (

=10). The treatment groups were administrated through gastric perfusion

and the normal group was given the equivalent volume of normal saline for 10 days. 1.5 h after the last treatment

chloral hydrate peritoneal anesthesia was performed

blood was collected from heart

and different doses of serum were separated to prepare drug-containing serum of low-dose

middle-dose

high-dose MSJZT groups

in order to incubate SGC-7901 gastric cancer cell. Early and late apoptosis rates were detected with flow cytometry. Afterwards

the tumor suppressor gene p53

c-nucleoprotein gene (c-Myc)

cysteine-aspartic acid protease-3 (Caspase-3)

B-cell lymphoma-2 (Bcl-2) mRNA expressions were confirmed by fluorescence quantitative polymerase chain reaction (Real-time PCR). The protein expressions of p53

c-Myc

Caspase-3

Bcl-2 were detected by immunofluorescence.

Result:

Compared with the normal group

the high-dose MSJZT group could obviously increase the apoptosis rate to 22.58%(

0.01). The results of Real-time PCR showed that the middle-dose MSJZT group significantly promoted the mRNA expression of Caspase-3 (

0.01)

the medium-dose group significantly increased the mRNA expressions of p53 and c-Myc (

0.01)

and the high-dose group significantly inhibited the mRNA expression of Bcl-2 (

0.01). The immunofluorescence results showed that the high-dose MSJZT group could significantly increase the protein expressions of c-Myc

Caspase-3 and p53 (

0.01)

while the protein expression of Bcl-2 was significantly reduced (

0.05).

Conclusion:

MSJZT drug serum could exert an anti-tumor effect by inhibiting the expression of the anti-apoptotic protein Bcl-2

and promoting the expressions of pro-apoptotic-related molecules p53

c-Myc

Caspase-3.

关键词

Keywords

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