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桦褐孔菌(Inonotus obliquus)因朵菌素提取物对人肺腺癌细胞系 A549 增殖周期和凋亡基因的影响。

Effects of inotodiol extracts from inonotus obliquus on proliferation cycle and apoptotic gene of human lung adenocarcinoma cell line A549.

机构信息

Department of Pathology, Jilin Medical College, Jilin, 132013, China.

出版信息

Chin J Integr Med. 2011 Mar;17(3):218-23. doi: 10.1007/s11655-011-0670-x. Epub 2011 Feb 27.

Abstract

OBJECTIVE

To observe the proliferation inhibition, apoptosis, and cell proliferation cycle of human lung carcinoma cell line A549 treated with Inotodiol extracts from Inonotus obliquus and explore the possibility of Inotodiol extracts from Inonotus obliquus as a new tumor chemopreventive drug.

METHODS

Human lung cancer cell line A549 was treated with different concentrations of Inotodiol, the effects of Inotodiol on cell apoptosis, the expression of Ki-67, Bcl-2, Bax, and p53 and cell cycle were detected by TUNEL assay, immunohistochemistry, and flow cytometry assay respectively.

RESULTS

Inotodiol extracts had antiproliferation effect on human lung carcinoma cell line A549. The expression of Ki-67 decreased with the increase of Inotodiol concentration and exposure time (P<0.05), in a dose-dependent and time-dependent manner. The typical characteristics of the apoptosis of A549 cells treated with Inotodiol were observed, and the apoptotic rate of A549 cell at 48 h was the highest by TUNEL assay. Inotodiol arrested A549 cells in the S phase, and apoptotic peak was observed by flow cytometry. Immunocytochemistry indicated that the expression of Bcl-2 protein decreased, while the expression of p53 and Bax proteins increased in A549 cells treated with Inotodiol, compared with the control cells (P<0.05).

CONCLUSION

Inotodiol can inhibit proliferation and induce the apoptosis of A549 cells, and its molecular mechanism may be associated with the up-regulating expression of p53 and bax proteins and down-regulating expression of Bcl-2 protein, which arrested A549 cells in S phase.

摘要

目的

观察桦褐孔菌醇提取物对人肺癌 A549 细胞的增殖抑制、凋亡及细胞周期的影响,探讨桦褐孔菌醇提取物作为新型肿瘤化学预防药物的可能性。

方法

用不同浓度的桦褐孔菌醇提取物处理人肺癌 A549 细胞,采用 TUNEL 法、免疫组化法和流式细胞术分别检测桦褐孔菌醇提取物对细胞凋亡、Ki-67 表达、Bcl-2、Bax 和 p53 蛋白表达及细胞周期的影响。

结果

桦褐孔菌醇提取物对人肺癌 A549 细胞具有增殖抑制作用。Ki-67 的表达随着桦褐孔菌醇浓度和作用时间的增加而降低(P<0.05),呈浓度和时间依赖性。TUNEL 法观察到桦褐孔菌醇处理 A549 细胞出现典型的凋亡特征,48 h 时 A549 细胞的凋亡率最高。桦褐孔菌醇将 A549 细胞阻滞在 S 期,流式细胞术观察到凋亡峰。免疫细胞化学法表明,桦褐孔菌醇处理 A549 细胞后 Bcl-2 蛋白表达降低,p53 和 Bax 蛋白表达增加(P<0.05)。

结论

桦褐孔菌醇能抑制 A549 细胞增殖并诱导其凋亡,其分子机制可能与上调 p53 和 bax 蛋白表达、下调 Bcl-2 蛋白表达,将 A549 细胞阻滞在 S 期有关。

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