Zhai Qiu-Li, Hu Xiang-Dan, Xiao Jing, Yu Dong-Qing
Second Clinical Medical College, Guangzhou University of Traditional Chinese Medicine， Guangzhou 510006， China.
Department of Gynecology, Guangdong Hospital of Traditional Chinese Medicine， Guangzhou 510006， China.
Zhongguo Zhong Yao Za Zhi. 2018 Feb;43(4):805-812. doi: 10.19540/j.cnki.cjcmm.20171113.018.
This study aimed to investigate the possible sensitivity of Astragalus polysaccharides, in order to improve the chemosensitivity of cervical cancer HeLa cells to cisplatin by regulating the cell autophagy, and explore its possible mechanism. In this study, HeLa cells were divided into control group, cisplatin group, Astragalus polysaccharide group, and Astragalus polysaccharide combined with cisplatin group. MTT assay was used to detect the proliferation of cervical cancer HeLa cells. Flow cytometry was used to detect the apoptosis and cycle of HeLa cells in each experimental group. RT-PCR was used to detect the mRNA expression of autophagy-related proteins beclin1, LC3Ⅱ and p62. The expression levels of autophagy-related proteins beclin1, LC3Ⅱ, LC3Ⅰ and p62 were detected by WB method. MTT results showed that compared with the control group, the proliferation of HeLa cells was significantly inhibited in each administration group(<0.05), and the inhibitory effect of the combination group was more significant(<0.01). The apoptotic rate of HeLa cells was significantly increased(<0.05), and the apoptotic rate of the combination group was significantly increased(<0.01) compared with the control group(<0.05).In conclusion, G₀/G₁ phase showed the most significant differences between the two groups. RT-PCR and WB results showed that the gene and protein expressions of beclin1 and LC3Ⅱ were up-regulated, while the gene and protein expressions of p62 were down-regulated compared with the control group. The above-mentioned changes in the combination group were more significant. Through the analysis of the above experimental results, it is speculated that Astragalus polysaccharides may increase the sensitivity of cervical cancer HeLa cells to cisplatin by regulating the cell autophagy. Its possible mechanism of action is correlated with the up-regulation of autophagy-related proteins beclin1, the promote the conversion from LC3Ⅰ to LC3Ⅱ, the down-regulation of labeled protein p62, and the enhancement of HeLa cell autophagic activity, thereby increasing the sensitivity of HeLa cells to cisplatin chemotherapy.
本研究旨在探讨黄芪多糖的可能敏感性，通过调节细胞自噬来提高宫颈癌HeLa细胞对顺铂的化疗敏感性，并探索其可能机制。本研究中，HeLa细胞分为对照组、顺铂组、黄芪多糖组和黄芪多糖联合顺铂组。采用MTT法检测宫颈癌HeLa细胞的增殖情况。采用流式细胞术检测各实验组HeLa细胞的凋亡及周期。采用RT-PCR法检测自噬相关蛋白beclin1、LC3Ⅱ和p62的mRNA表达。采用WB法检测自噬相关蛋白beclin1、LC3Ⅱ、LC3Ⅰ和p62的表达水平。MTT结果显示，与对照组相比，各给药组HeLa细胞的增殖均受到显著抑制（<0.05），联合组的抑制作用更显著（<0.01）。HeLa细胞的凋亡率显著升高（<0.05），联合组的凋亡率与对照组相比显著升高（<0.01）。总之，G₀/G₁期两组间差异最为显著。RT-PCR和WB结果显示，与对照组相比，beclin1和LC3Ⅱ的基因和蛋白表达上调，而p62的基因和蛋白表达下调。联合组的上述变化更为显著。通过对上述实验结果的分析推测，黄芪多糖可能通过调节细胞自噬增加宫颈癌HeLa细胞对顺铂的敏感性。其可能的作用机制与自噬相关蛋白beclin1的上调、促进LC3Ⅰ向LC3Ⅱ的转化、标记蛋白p62的下调以及HeLa细胞自噬活性的增强有关，从而增加HeLa细胞对顺铂化疗的敏感性。
