荷叶富含类黄酮提取物通过调控 PI3K 相关分子表达抑制 HT-29 结肠癌细胞生长
Inhibitory Effect of Lotus Leaf-Enriched Flavonoid Extract on the Growth of HT-29 Colon Cancer Cells through the Expression of PI3K-Related Molecules.
机构信息
Chongqing Collaborative Innovation Center for Functional Food, Chongqing University of Education, Chongqing 400067, China.
Chongqing Engineering Research Center of Functional Food, Chongqing University of Education, Chongqing 400067, China.
出版信息
Biomed Res Int. 2022 May 9;2022:6770135. doi: 10.1155/2022/6770135. eCollection 2022.
OBJECTIVES
Lotus leaf is rich in flavonoids, and this study is aimed at examining the inhibitory effect of lotus leaf-enriched flavonoid extract (LLEFE) on HT-29 colon cancer cells through phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) expression regulation.
METHODS
Lotus leaves were extracted by ethanol and purified using FL-3 macroporous resin to create the LLEFE. HT-29 colon cancer cells were tested using various methods: their proliferation was observed by 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay, their survival status was observed by fluorescence staining, their oxidative stress level was observed by biochemical kits, and their mRNA expression was determined by quantitative polymerase chain reaction (qPCR) assay. Additionally, the composition of the flavonoids in lotus leaf was determined by HPLC.
RESULTS
The results showed that the proliferation of NCM460 normal human colon cells was not affected by 0-500 g/mL LLEFE but the proliferation of HT-29 human colon cancer cells decreased. LLEFE increased the LDH level in an HT-29 colon cancer cell culture medium; also increased the superoxide dismutase (SOD), catalase (CAT) activities, and glutathione (GSH) level in HT-29 cells; and decreased the malondialdehyde (MDA) level. Further experimental results showed that LLEFE upregulated the expression of , , and mRNA and downregulated the expression of , , and in HT-29 cells. The high-performance liquid chromatography (HPLC) results showed that kaempferin, hyperoside, astragaloside, phloridzin, and quercetin were the main chemical constituents of lotus leaf.
CONCLUSION
Lotus leaves contain functional flavonoids that inhibit the proliferation of HT-29 colon cancer cells and regulate the expression of PI3K/Akt through five important chemicals.
目的
荷叶富含类黄酮，本研究旨在通过调节磷脂酰肌醇 3-激酶/蛋白激酶 B（PI3K/Akt）表达来研究荷叶富含类黄酮提取物（LLEFE）对 HT-29 结肠癌细胞的抑制作用。
方法
采用乙醇提取荷叶，用 FL-3 大孔树脂纯化得到 LLEFE。采用多种方法检测 HT-29 结肠癌细胞：通过 3-(4,5)-二甲基噻唑(-z-y1)-3,5-二苯基四氮唑溴盐（MTT）比色法观察细胞增殖，荧光染色观察细胞存活状态，生化试剂盒观察细胞氧化应激水平，定量聚合酶链反应（qPCR）检测细胞 mRNA 表达。此外，采用高效液相色谱法（HPLC）测定荷叶中类黄酮的组成。
结果
结果表明，0-500μg/mL LLEFE 对 NCM460 正常结肠细胞的增殖没有影响，但 HT-29 人结肠癌细胞的增殖减少。LLEFE 增加了 HT-29 结肠癌细胞培养物中乳酸脱氢酶（LDH）的水平；增加了 HT-29 细胞中超氧化物歧化酶（SOD）、过氧化氢酶（CAT）的活性和谷胱甘肽（GSH）水平；降低了丙二醛（MDA）的水平。进一步的实验结果表明，LLEFE 上调了 HT-29 细胞中 、 、 mRNA 的表达，下调了 、 、 mRNA 的表达。高效液相色谱（HPLC）结果表明，山奈酚、金丝桃苷、黄芪甲苷、根皮苷和槲皮素是荷叶的主要化学成分。
结论
荷叶含有具有功能的类黄酮，可抑制 HT-29 结肠癌细胞的增殖，并通过五种重要化学物质调节 PI3K/Akt 的表达。
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