仙鹤草通过诱导肿瘤细胞凋亡和自噬抑制肝癌裸鼠移植瘤生长

Agrimonia Pilosa Ledeb Inhibits the Growth of Transplanted Hepatoma in Nude Mice by Inducing Tumor Cell Apoptosis and Autophagy

目的探究仙鹤草(APL)对肝癌裸鼠移植瘤的抑制作用及其机制。方法裸鼠皮下注射肝癌Hep G2细胞,建立裸鼠移植瘤模型;将裸鼠随机分为5组:对照组(Control)、仙鹤草低剂量组(APL 6g/kg)、仙鹤草中剂量组(APL 12g/kg)、仙鹤草高剂量组(APL 24g/kg)和5-氟尿嘧啶组(5-FU 20mg/kg)。30d后检测肿瘤重量;检测30d期间肿瘤体积变化;TUNEL检测肿瘤组织凋亡;Western blot检测Caspase-3、Bax、Bcl-2、LC3-I、LC3-II、Beclin1、p62蛋白表达水平;免疫组化染色检测Ki67、VEGF阳性细胞数。结果与Control组比较,APL各剂量组和5-FU 20mg/kg组裸鼠肿瘤重量、体积显著降低,裸鼠肿瘤组织细胞凋亡率显著升高,裸鼠肿瘤组织细胞凋亡率显著升高,Bcl-2蛋白水平显著降低,LC3-II/LC3-I比值和Beclin1蛋白水平显著升高,p62蛋白水平显著降低,Ki67、VEGF阳性细胞数显著降低,差异有统计学意义(P<0.05或P<0.01)。结论仙鹤草抑制肝癌裸鼠移植瘤的生长,可能参与机制为诱导肿瘤细胞凋亡和自噬,抑制肿瘤细胞增殖。

Objective To investigate the inhibitory effect of Agrimonia Pilosa Ledeb(APL)on transplanted tumor of liver cancer in nude mice and its mechanism.Methods Hep G2 cells were subcutaneously injected into nude mice,which were randomly divided into 5 groups:Control group,Agrimonia Pilosa Ledeb low-dose group(APL 6g/kg),Agrimonia Pilosa Ledeb medium-dose group(APL 12g/kg),Agrimonia Pilosa Ledeb high-dose group(APL 24g/kg)and 5-fluorouracil group(5-FU 20mg/kg).The tumor weight was detected 30 days later.The changes of tumor volume at 30 days were detected.TUNEL was used to detect tumor apoptosis.The protein expression levels of Caspase-3,Bax,Bcl-2,LC3-I,LC3-II,Beclin1 and p62 were detected by Western blot.Ki67 and VEGF positive cells were detected by immunohistochemical staining.Results Compared with the Control group,the weight and volume of tumor were significantly decreased in APL dose groups and 5-Fu 20mg/kg group,and the apoptosis rate of tumor tissue was significantly increased,and the protein level of Bcl-2 was significantly decreased.LC3-II/LC3-I ratio and Beclin1 protein level were significantly increased,p62 protein level was significantly decreased,Ki67 and VEGF positive cell number were significantly decreased,the differences were statistically significant(P<0.05 or P<0.01).Conclusion APL may be involved in inducing apoptosis and autophagy of tumor cells and inhibiting proliferation of tumor cells by inhibiting the growth of transplanted tumor in nude mice.