Effects of Safflower Polysaccharide on Gene Transcription and Protein Express on of Bcl-2 and Bax in Human Hepatocarcinoma Cell Line SMMC-7721

ZHANG Xiao-li; CHENG Xiang; LIU Yang; SHI Xue-kui

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Effects of Safflower Polysaccharide on Gene Transcription and Protein Express on of Bcl-2 and Bax in Human Hepatocarcinoma Cell Line SMMC-7721

更新时间：2024-01-04

- Effects of Safflower Polysaccharide on Gene Transcription and Protein Express on of Bcl-2 and Bax in Human Hepatocarcinoma Cell Line SMMC-7721
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 18, Issue 14, Pages: 239-244(2012)
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- Published：2012
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ZHANG Xiao-li, CHENG Xiang, LIU Yang, et al. Effects of Safflower Polysaccharide on Gene Transcription and Protein Express on of Bcl-2 and Bax in Human Hepatocarcinoma Cell Line SMMC-7721[J]. Chinese journal of experimental traditional medical formulae, 2012, 18(14): 239-244.
DOI：

ZHANG Xiao-li, CHENG Xiang, LIU Yang, et al. Effects of Safflower Polysaccharide on Gene Transcription and Protein Express on of Bcl-2 and Bax in Human Hepatocarcinoma Cell Line SMMC-7721[J]. Chinese journal of experimental traditional medical formulae, 2012, 18(14): 239-244. DOI：

摘要

目的: 研究红花多糖(SPS)体外抑制人肝癌细胞株SMMC-7721增殖、诱导凋亡及对凋亡调控基因Bax

Bcl-2基因转录和蛋白表达的影响

探讨SPS诱导SMMC-7721细胞凋亡的机制。方法: 不同剂量的SPS(0

0.02

0.04

0.08

0.16

0.32

0.64

1.28 g·L-1)分别作用于体外培养的SMMC-7721细胞

采用四甲基偶氮唑盐比色法(MTT)检测细胞的抑制率;用Ca2+依赖性磷脂结合蛋白(AnnexinⅤ)-异硫氰酸荧光素(FITC)/碘化丙啶(PI) 双染法观察细胞凋亡的形态学改变;通过罗丹明123(Rho123)荧光显微镜检测线粒体膜电位(Δψm);以实时荧光定量 PCR 技术 (real-time PCR) 和蛋白免疫印迹 (Western blotting) 法检测细胞凋亡相关因子 Bcl-2及Bax mRNA 水平和蛋白水平的表达情况。结果: SPS对体外培养的人肝癌SMMC-7721细胞具有抑制增殖作用

且具有明显的时间和剂量相关性;荧光显微镜下SPS作用的细胞呈现典型的凋亡细胞形态;肝癌细胞内Rho123荧光强度明显减弱。SPS作用后细胞内Bcl-2蛋白和mRNA表达水平均降低

Bax蛋白和mRNA的表达水平均升高

二者的变化趋势均呈一定的时间依赖关系;而且

Bcl-2/Bax比值随SPS时间的延长显著降低且变化趋势均呈一定的时间依赖关系。结论: SPS能够显著抑制人肝癌细胞SMMC-7721增殖

诱导SMMC-7721细胞凋亡

其作用机制可能与上调Bax的表达及下调Bcl-2的表达和降低线粒体膜电位有关。

Abstract

Objective: To explore how safflower polysaccharide (SPS) induce apoptosis of human liver cancer cell line SMMC-7721 through observing the influence of SPS on proliferation

apoptosis and expression of apoptosis gene and protein Bax and Bcl-2 of SMMC-7721. Method: SMMC-7721 cells were treated with different concentrations(0

0.02

0.04

0.08

0.16

0.32

0.64

1.28 g·L-1) of SPS. The cells growth inhibitory rate was measured by methyl thiazolyl tetrazolium(MTT) assay. The morphological changes of apoptosis were observed by AnnexinⅤ-FITC/PI staining. The mitochondrial membrane potential(Δψm) was assayed by fluorescent microscopy using Rhodamine 123(Rho123). Quantitative real-time RT-PCR and Western blot analysis were used to detect the mRNA and protein expressions of Bcl-2 and Bax. Result: SPS could inhibit the proliferation of SMMC-7721 cells in a dose and time dependent manner. After treated with SPS

SMMC-7721 cells presented typical apoptosis under fluorescence microscope.Fluorescence intensity of Rho123 were significantly decreased in SMMC-7721 cells. The mRNA and protein expression of Bcl-2 expression were down-regulated while those of Bax were up-regulated and both changes had good time-dependent tendency. The ratio of Bcl-2/Bax decreased significantly in a time-dependent manner. Conclusion: SPS could inhibit the proliferation of SMMC-7721 and enhance apoptosis of SMMC-7721 and its mechanism maybe relate with its up-regulation of Bax expression as well as down-regulation of Bcl-2 expression and decline of Δψm.

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