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[转基因雪莲黄酮类化合物及其抗肿瘤活性分析]

[Analysis of flavonoids and antitumor activity of transgenic Saussurea involucrate].

作者信息

Cao Zhi-Xiu, Li Hou-Hua, Li Ai, Liu Peng-Yuan, Zhao Yan-Ying, Mao Pei-Qi, Li Gui-Lin

机构信息

College of Landscape Architecture and Arts, Northwest A & F University, Yangling 712100, China.

Nanjing International Medical Technology Services Limited, Nanjing 210000, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2018 Nov;43(22):4491-4497. doi: 10.19540/j.cnki.cjcmm.20180619.001.

Abstract

The aim of this paper was to investigate the flavonoids of callus of transgenic and non-transgenic Saussurea involucrate and its antitumor activity on the esophageal cancer CaEs-17 cells. The species and content of mono-phenols were detected by high performance liquid chromatography. The growth of human esophageal cancer CaEs-17 cells was detected using CCK-8 assay, apoptosis morphology observation and flow cytometry. Expression of related apoptotic genes Bax and Bcl-2 were determined by qPCR. The results showed that the content of total flavonoids in the transgenic callus was 2.72 times that of the non-transgenic callus. The cyanidin-galactoside was detected in the transgenic callus, but not in the non-transgenic callus. The inhibitory effect of the extracts from the transgenic callus on CaEs-17 cells was more significant than that of the non-transgenic callus, and the IC₅₀ value had a decreased of 26.4%. Flow cytometry analysis results showed that the apoptosis induction effect of the extracts from the transgenic callus on CaEs-17 cells was significantly better than that of the non-transgenic callus. Fluorescence quantitative PCR analysis results showed that the extracts from the transgenic callus could up-regulate the expression of proapoptotic gene Bax and down-regulate the expression of apoptotic gene Bcl-2, and the regulation effect of the transgenic callus was more significant. Therefore, compared with the non-transgenic callus, the antitumor activity of the extracts from the callus of transgenic S. involucrate on the esophageal cancer CaEs-17 cells was significantly increased, which was closely related to the accumulation of cyanidin-galactoside and its metabolism-related flavonoid compounds in the transgenic callus.

摘要

本文旨在研究转基因和非转基因雪莲愈伤组织中的黄酮类化合物及其对食管癌CaEs - 17细胞的抗肿瘤活性。采用高效液相色谱法检测单酚类的种类和含量。使用CCK - 8法、凋亡形态观察和流式细胞术检测人食管癌CaEs - 17细胞的生长情况。通过qPCR测定相关凋亡基因Bax和Bcl - 2的表达。结果表明,转基因愈伤组织中总黄酮含量是非转基因愈伤组织的2.72倍。在转基因愈伤组织中检测到矢车菊素 - 半乳糖苷,而在非转基因愈伤组织中未检测到。转基因愈伤组织提取物对CaEs - 17细胞的抑制作用比非转基因愈伤组织更显著,IC₅₀值降低了26.4%。流式细胞术分析结果表明,转基因愈伤组织提取物对CaEs - 17细胞的凋亡诱导作用明显优于非转基因愈伤组织。荧光定量PCR分析结果表明,转基因愈伤组织提取物可上调促凋亡基因Bax的表达,下调凋亡基因Bcl - 2的表达,且转基因愈伤组织的调控作用更显著。因此,与非转基因愈伤组织相比,转基因雪莲愈伤组织提取物对食管癌CaEs - 17细胞的抗肿瘤活性显著增强,这与转基因愈伤组织中矢车菊素 - 半乳糖苷及其代谢相关黄酮类化合物的积累密切相关。

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