Liu Ze-Hong, Chen Di-Long, Jiang Rong, Chen Yi, Xiong Wei, Wang Fen, Shi Xue-Ping, Li Hai-Xing, Li Jing
Laboratory of Stem Cell and Tissue Engineering, Chongqing Medical University, Chongqing 400016, China.
Chongqing Three Gorges Medical College, Chongqing 402120, China.
Zhongguo Zhong Yao Za Zhi. 2016 Feb;41(4):700-704. doi: 10.4268/cjcmm20160426.
To study the in vivo inhibition effect of ginsenoside Rh₂ on humanleukemia cells, and explore its mechanism from autophagy and apoptosis aspects, human leukemia K562 cells allograft tumor models were applied, and after administration of ginsenosides Rh₂ by gavage, the tumor diameter, volume and inhibitory rate were measured, and the anti-tumor activity of ginsenosides Rh₂ was observed. The levels of HAT and HDAC in tumor tissues were detected by chemical colorimetry assay, and expressions of HDAC1, HDAC2, HDAC3, HDAC4, HDAC5 and HDAC6 were detected by Western blotting assay. The expression levels of vital genes closely associated with autophagy and mRNA expressions of HDAC6 and Hsp90 were detected by Real time-PCR. HE staining was used to observe apoptosis, and immunohistochemistry was used to detect the protein expressions of HDAC6, Hsp90 and activated caspases 3. The results showed that ginsenoside Rh₂ could inhibit the growth of k562 cells allograft tumor, with a tumor inhibition rate up to 53.10%. Ginsenoside Rh₂ could significantly decrease HDAC activity and decrease the expressions of HDAC1, HDAC2 and HDAC6, and inhibit the expressions of HDAC6 and HSP90, increase the expressions of vital autophagy genes (beclin-1, LC3A and LC3B). Histopathological results showed that ginsenosides Rh₂ could significantly increase the tumor apoptosis. Therefore, ginsenoside Rh₂ had good anti-tumor effect in vivo, and the mechanism maybe associated with regulating autophagy and apoptosis through HDAC6 and Hsp90 pathways and inhibiting the in vivo proliferation of tumor cells.
为研究人参皂苷Rh₂对人白血病细胞的体内抑制作用，并从自噬和凋亡方面探讨其作用机制，应用人白血病K562细胞异体移植瘤模型，经灌胃给予人参皂苷Rh₂后，测量肿瘤直径、体积及抑制率，观察人参皂苷Rh₂的抗肿瘤活性。采用化学比色法检测肿瘤组织中组蛋白乙酰转移酶（HAT）和组蛋白去乙酰化酶（HDAC）水平，采用蛋白质印迹法检测HDAC1、HDAC2、HDAC3、HDAC4、HDAC5和HDAC6的表达。采用实时荧光定量PCR检测与自噬密切相关的关键基因表达水平以及HDAC6和热休克蛋白90（Hsp90）的mRNA表达。采用苏木精-伊红（HE）染色观察细胞凋亡情况，采用免疫组织化学法检测HDAC6、Hsp90和活化的半胱天冬酶3的蛋白表达。结果显示，人参皂苷Rh₂可抑制K562细胞异体移植瘤生长，肿瘤抑制率高达53.10%。人参皂苷Rh₂可显著降低HDAC活性，减少HDAC1、HDAC2和HDAC6的表达，抑制HDAC6和Hsp90的表达，增加关键自噬基因（beclin-1、LC3A和LC3B）的表达。组织病理学结果显示，人参皂苷Rh₂可显著增加肿瘤细胞凋亡。因此，人参皂苷Rh₂在体内具有良好的抗肿瘤作用，其机制可能与通过HDAC6和Hsp90途径调节自噬和凋亡以及抑制肿瘤细胞的体内增殖有关。
