Effects of  on Proliferation,Apoptosis and Expression of VEGF,MMP-2 of SMMC 7721 Cells

ZENG Zhi-tao; FU Ying; XIONG Yao-bin; ZI Xiao-fei; PENG Zhong-juan; HUANG Ling-ling; YANG Hui; YU Hong

doi:10.13422/j.cnki.syfjx.2015090141

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Effects of on Proliferation,Apoptosis and Expression of VEGF,MMP-2 of SMMC 7721 Cells

更新时间：2024-01-04

- Effects of on Proliferation,Apoptosis and Expression of VEGF,MMP-2 of SMMC 7721 Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 21, Issue 9, Pages: 141-144(2015)
- 作者机构：
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.2015090141
  CLC： R285.5
- Published：2015
- 稿件说明：
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ZENG Zhi-tao, FU Ying, XIONG Yao-bin, et al. Effects of on Proliferation,Apoptosis and Expression of VEGF,MMP-2 of SMMC 7721 Cells[J]. Chinese journal of experimental traditional medical formulae, 2015, 21(9): 141-144.
DOI：

ZENG Zhi-tao, FU Ying, XIONG Yao-bin, et al. Effects of on Proliferation,Apoptosis and Expression of VEGF,MMP-2 of SMMC 7721 Cells[J]. Chinese journal of experimental traditional medical formulae, 2015, 21(9): 141-144. DOI： 10.13422/j.cnki.syfjx.2015090141.

摘要

目的: 探讨从天然冬虫夏草中分离出的虫草拟青霉Cs-4菌株

经人工发酵所得发酵虫草菌粉对SMMC-7721人肝癌细胞的生长抑制作用. 方法: 20只体重(200±20)g雄性SD大鼠随机分为4组(每组5只):空白组及Cs-4低、中、高剂量组

空白组ig予蒸馏水

Cs-4低、中、高剂量组(0.77

1.54

3.08 g·kg-1

ig);给药3 d后

制备含药血清.肝癌细胞悬液调整至4×104/mL

96孔板每孔加入100 μL

培养24 h后加入大鼠含药血清及顺铂注射液;干预48 h MTT法测各组细胞增殖抑制率.将SMMC-7721细胞悬液调整至2.8×104/mL

接种于6孔板培养24 h

加入大鼠含药血清及顺铂

干预培养48 h后Annexin V-FITC/PI染色

流式细胞仪检测各组细胞凋亡率;ELISA检测各组细胞上清液中血管内皮生长因子(VEGF)

金属基质蛋白酶-2(MMP-2)表达量. 结果: SMMC-7721细胞增殖被Cs-4含药血清明显抑制(与空白组比较P< 0.05)

低、中、高剂量组抑制率分别为12.2%

23.2%

38.1%

呈剂量依赖性(Cs-4不同剂量组间比较P< 0.05).Cs-4含药血清低、中、高剂量组细胞凋亡率分别为13.9%

23.5%

24.9%

均高于空白组(P< 0.05).Cs-4含药血清低、中、高剂量组细胞上清液中VEGF分别为443.5

436.9

411.0 ng·L-1

低于空白组(P< 0.05);Cs-4低、中、高剂量组细胞上清液中MMP-2浓度与空白组比无显著差异. 结论: Cs-4含药血清能抑制SMMC-7721细胞增殖

提高体外培养的SMMC-7721细胞凋亡率

抑制SMMC-7721细胞VEGF表达

提示Cs-4有抗肝癌作用

对SMMC-7721细胞MMP-2表达无显著影响.

Abstract

Objective: To explore the effect of the Chinese caterpillar fungus isolated from natural Paecilomyces hepiali strain (Cs-4) inhibited by artificial fermentation

the Fermented-4 powder on SMMC-7721 human hepatocellular carcinoma cell growth. Method: Twenty male SD rats were randomly divided into four groups (n=5):control group and Cs-4 low

medium and high dose groups

control group was treated with distilled water

Cs-4 low

middle and high dose groups were treated with distilled water to prepare the Cs-4 (0.77

1.54

3.08 g·kg-1

ig)suspension

according to the 10 mL·kg-1 body weight orallyorally

2 times twice a daily;after intragastric administrationfor 3 days

the serum containing drug was prepared. The liver cell suspension was adjusted to 4×104 cells/mL

each hole of 96 hole plate was added with 100 μL cell suspension culture. After 24 h

the holes were respectively added with 160 μL cells culture fluid and 40 μL blank serum and containing Cs-4.After 48 h MTT detection was carried out

the inhibition rate of proliferation of cells was calculated. The SMMC-7721 cell suspension was adjusted to 2.8×104 cells/mL

were cultured in 6 well plates (each hole 2 mL inoculation) for 24 h

the holes were added to 1.6 mL cells suspension and 0.4 mL black serum and containing drug serum were added after 48 h

Annexin V-FITC/PI staining by flow cytometry was used to detect of apoptosis;expression of vascular endothelial growth factor(VEGF)

matrix metalloproteinases-2(MMP-2) were detected with ELISA method. Result: MTT showed the proliferation of SMMC-7721 cells was significantly inhibited by containing Cs-4 serum compared with the blank control group(P <0.05)

the different dose inhibition rates were 12.2%

23.2%

38.1% in a dose-dependent manner. Apoptosis detection showed that in containing Cs-4 serum groups

the apoptosis rates were 13.9%

23.5%

24.9%

which was higher than those in the blank group (P <0.05). In containing Cs-4 serum groups

the concentration of VEGF in supernatant was respectively 443.5

436.9

411.0 ng·L-1

which was lower than that in control group (P <0.05). Conclusion: The containing Cs-4 serum can inhibit the proliferation and VEGF expression of SMMC-7721 cells

and increase the apoptosis of SMMC-7721 suggesting Cs-4 had anti-tumor metastasis action.

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