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姜黄素诱导大肠癌细胞LoVo凋亡的作用及机制研究

[Study on functions and mechanism of curcumin in inducing colorectal carcinoma cells LoVo apoptosis].

作者信息

Guo Li-Da, Jiao Zhen-Xia, Song Ying, Teng Wen-Hua, Liu Zhao, Liu Jing-Ze

机构信息

Department of Environment and Chemical Engineering, Hebei College of Industry and Technology, Shijiazhuang 050091, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2013 Jul;38(13):2191-6.

Abstract

OBJECTIVE

To discuss the biological function and regulation mechanism of curcumin in promoting human colorectal carcinoma (LoVo) cells apoptosis.

METHOD

Conventional in vitro culture in human colorectal carcinoma cells LoVo, When 80%-90% confluence was reached, cells were treated with curcumin at different concentrations (0-20 mg x L(-1)). Curcumin's effect on cell proliferation level was examined by MTT colorimetry. The ultrastructure of curcumin-treated LoVo cells were observed with transmission electron microscope (TEM). The amount of PI-positive LoVo cells after the curcumin treatment were determined by flow cytometry. The cell apoptosis rate was detected by Annexin V-FITC/PI double staining. The mRNA level of Bax, Bcl-2, Caspase-3 and Bcl-xL were tested by means of RT-PCR.

RESULT

MTT test indicates curcumin could inhibite the growth and proliferation of LoVo cells in a time- and concentration-dependent manner. TEM examination showed that curcumin can make LoVo cell morphological changes, showing the typical characteristics of apoptotic cells. Flow cytometry instrument analysis showed that curcumin can arrest cell cycle at S phase, and induce apoptosis of LoVo cells. RT-PCR test showed that curcumin can activate the expression of Bax and Caspase-3, inhibit the expression of Bcl-2 and Bcl-xL at the mRNA level.

CONCLUSION

Curcumin can significantly inhibit the proliferation and induce the apoptosis of human colorectal carcinoma cells LoVo. Such biological effect may be associated with activating Caspase-3 signal channel by activating Bax expression and inhibiting Bcl-2 and Bcl-xL expression. This study lays an important foundation for further discussing the mechanism of curcumin in inducing human colorectal carcinoma LoVo apoptosis.

摘要

目的

探讨姜黄素促进人结肠癌细胞(LoVo)凋亡的生物学功能及调控机制。

方法

采用人结肠癌细胞LoVo常规体外培养,待细胞达到80%-90%汇合时,用不同浓度(0-20mg·L⁻¹)的姜黄素处理细胞。采用MTT比色法检测姜黄素对细胞增殖水平的影响。用透射电子显微镜(TEM)观察经姜黄素处理的LoVo细胞的超微结构。通过流式细胞术测定姜黄素处理后PI阳性LoVo细胞的数量。采用Annexin V-FITC/PI双染法检测细胞凋亡率。通过RT-PCR检测Bax、Bcl-2、Caspase-3和Bcl-xL的mRNA水平。

结果

MTT试验表明,姜黄素能以时间和浓度依赖性方式抑制LoVo细胞的生长和增殖。TEM检查显示,姜黄素可使LoVo细胞形态发生改变,呈现凋亡细胞的典型特征。流式细胞仪分析表明,姜黄素可使细胞周期阻滞于S期,并诱导LoVo细胞凋亡。RT-PCR试验表明,姜黄素在mRNA水平上可激活Bax和Caspase-3的表达,抑制Bcl-2和Bcl-xL的表达。

结论

姜黄素可显著抑制人结肠癌细胞LoVo的增殖并诱导其凋亡。这种生物学效应可能与通过激活Bax表达、抑制Bcl-2和Bcl-xL表达来激活Caspase-3信号通道有关。本研究为进一步探讨姜黄素诱导人结肠癌LoVo细胞凋亡的机制奠定了重要基础。

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