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目的观察叶下珠复方Ⅱ号对肝癌HepG_2细胞裸鼠移植瘤生长的抑制作用,并探讨其作用机制。方法建立BALB/C裸鼠肝癌HepG_2细胞皮下移植瘤模型,随机分为模型组,叶下珠复方Ⅱ号高、中、低剂量组(17.28,8.64,4.32 g·kg~(-1))和氟尿嘧啶(5-Fu)组(0.025 g·kg~(-1)),给药6周后处死裸鼠,剥离瘤组织称质量。体外观察叶下株复方Ⅱ号对HepG_2细胞增殖、集落形成、细胞周期和细胞凋亡的影响,观察药物作用后Wnt1和β-catenin基因mRNA表达的改变。结果叶下珠复方Ⅱ高、中、低剂量组对裸鼠HepG_2肝癌细胞移植瘤生长的抑制作用明显,与模型组比较,肿瘤质量均明显减轻(P<0.01),但与5-Fu组比较,差异均无统计学意义(P>0.05)。叶下珠复方Ⅱ号体外在10 mg·mL~(-1)以上浓度能明显抑制肝癌Hep G2细胞的增殖,呈明显的量-效关系和时-效关系;叶下珠复方Ⅱ号在2 mg·mL~(-1)以上浓度,对HepG_2细胞集落形成呈明显的抑制作用;叶下珠复方Ⅱ号以15,20 mg·mL~(-1)浓度作用HepG_2细胞48 h后,G0/G1期细胞比例明显降低,S期细胞比例显著增加,与细胞对照组比较,差异均具有统计学意义(P<0.01);叶下珠复方Ⅱ号在10,15,20 mg·mL~(-1)浓度,作用于HepG_2细胞48 h,细胞早期凋亡率和总凋亡率明显增加,与细胞对照组比较,差异均具有统计学意义(P<0.01)。叶下珠复方Ⅱ号高、低剂量组(20,10 mg·mL~(-1))作用于HepG_2细胞48 h后,Wnt1和β-catenin基因m RNA的表达量均明显低于细胞对照组(P<0.01,P<0.05)。结论叶下珠复方Ⅱ号对BALB/C裸鼠肝癌HepG_2细胞皮下移植瘤的生长具有明显的抑制作用,作用机制与抑制Wnt1/β-catenin基因表达和信号通路活化,从而阻滞细胞周期、抑制细胞增殖和诱导细胞凋亡有关。
Objective To observe the inhibitory effect of Ye Xia Zhu Fu Fang No.2 on the growth of HepG2 hepatoma xenografts in nude mice and to explore its mechanism. Methods BALB / C nude mice bearing HepG2 cells were established and subcutaneously transplanted into nude mice. The models were randomly divided into model group, high, medium and low doses of Phyllanthus Compound Ⅱ (17.28,8.64,4.32 g · kg -1) Fluorouracil (5-Fu) group (0.025 g · kg -1). After 6 weeks of administration, the nude mice were sacrificed and the tumor tissue was dissected and weighed. The effects of Yexia Zhuang Fufang Ⅱ on proliferation, colony formation, cell cycle and apoptosis of HepG2 cells were observed in vitro. The changes of mRNA expression of Wnt1 and β-catenin were observed. Results Phyllanthus Compound Ⅱ high, medium and low dose group of HepG 2 hepatoma cell transplanted tumor growth inhibition significantly, compared with the model group, the tumor mass were significantly reduced (P <0.01), but compared with the 5-Fu group , There was no significant difference (P> 0.05). Phyllanthus Compound Ⅱ could significantly inhibit the proliferation of Hep G2 cells in vitro at a concentration above 10 mg · mL -1, showing a dose-response and time-effect relationship. mg · mL -1 could significantly inhibit HepG_2 cell colony formation. After treated with Phyllanthus Compound Ⅱ at HepG_2 cell concentration of 15 and 20 mg · mL -1, G0 / The proportion of cells in G1 phase was significantly decreased and the proportion of cells in S phase was significantly increased (P <0.01). Compared with the cell control group, the difference was statistically significant (P <0.01) 1) concentration for 48 h, the early apoptosis rate and total apoptosis rate of HepG_2 cells increased significantly. Compared with the cell control group, the differences were statistically significant (P <0.01). The mRNA expression of Wnt1 and β-catenin in HepG_2 cells treated with Phyllanthus Compound II at high dose and low dose (20, 10 mg · mL -1) for 48 h were significantly lower than those in control cells (P <0.01, P <0.05). Conclusions Yexiazhu Compound Ⅱ significantly inhibits the growth of subcutaneously transplanted hepatocellular carcinoma HepG_2 cells in BALB / c nude mice by inhibiting the expression of Wnt1 / β-catenin and signaling pathway, thus blocking cell cycle and inhibiting Cell proliferation and induction of apoptosis related.