齐墩果酸对肝癌HCCLM3细胞裸鼠移植瘤的抑制作用及机制研究

Inhibitory effect of oleanolic acid on liver cancer HCCLM3 cells transplanted tumor in nude mice and its mechanism

目的研究齐墩果酸对人肝癌HCCLM3细胞裸鼠移植瘤生长的影响,并探讨其对肿瘤组织凋亡及相关基因表达的影响。方法BALB/c裸鼠皮下接种人肝癌细胞株HCCLM3构建肝癌移植瘤模型,随机分为模型组和药物组,分别给予0.01 mL/g玉米油和5 mg/kg齐墩果酸灌胃。给药期间测量并记录各组裸鼠的一般状态、体质量、瘤体质量和体积。通过HE、Ki67、TUNEL染色分别检测移植瘤组织中细胞坏死病理学改变、细胞增殖、细胞凋亡情况;采用qRT‐PCR检测移植瘤组织中CyclinD1、CyclinE1、CDK4、Bcl⁃2和Bax基因的mRNA表达水平。结果与模型组相比,药物组裸鼠体质量无明显差异,移植瘤体质量和体积均显著下降(均P<0.05)。HE染色结果显示药物组移植瘤组织形态发生明显改变,出现坏死区域。药物组瘤组织Ki67阳性表达强度显著低于模型组(P<0.01);TUNEL阳性表达强度显著高于模型组(P<0.05)。齐墩果酸给药后移植瘤组织中增殖相关基因CyclinD1、CyclinE1和CDK4的mRNA表达水平均降低,但差异无统计学意义(均P>0.05);抑凋亡相关基因Bcl⁃2的mRNA表达水平降低(P<0.05),而促凋亡相关基因Bax的mRNA表达水平升高(P<0.01)。结论齐墩果酸能抑制人肝癌HCCLM3细胞裸鼠移植瘤生长,其机制可能与抑制细胞增殖和诱导细胞凋亡相关。

Objective To investigate the effects of oleanolic acid on the growth of human liver cancer HCCLM3 cells transplanted tumor in nude mice,the apoptosis of tumor tissues and the expression of related genes.Methods Human liver cancer HCCLM3 cells were inoculated into the subcutancous skin of BALB/e nude mice to construct liver cancer transplantation tumor model,and the nude mice were randomly divided into model group and dnug group,which were given 0.01 ml/g com oil and 5 mg/kg oleanolic acid by gavage.respectively.The general state,body weight,tumor weight and volume of nude mice in each group wеre measured and recorded during administration.The pathological changes of cell necosis,cell proliferation and apoptosis in transplanted tumor tissues were detected by HE,Ki67 and TUNEL.staining,respectively.The mRNA expression levels of CyclinDI,CyclinEI,CDK4,Bcl-2 and Bax genes in transplanted tumor tissues were detected by qRT-PCR.Results Compared with the model group,there was no significant effect on the body weight of mude mice in the drug group,while the weight and volume of transplanted tumor were significantly decreased(all P<0.05).HE staining showed that the morphology of transplanted tumor in the drug group changed significantly,and necrotic areas appeared.The positive expression intensity of Ki67 in tumor tissues of the drug group was significantly lower than thatøf the model group(Pc0.01);the positiveexpression intensity of TUNEL.was significantly higher than that of the model group(P<c0.0S).The mRNA expression levels of proliferation-related genes CyclinDI,CyclinE1 and CDK4 intransplanted tumortissnes were decreased afteroleanolicacidadministration,but the differences were not statistically significant(all P>0.05);the mINA expression level of anti-apoptosis-related gene Bcl-2 was decreased(Pc0.05).while the mRNA expression level of pro-apoptosis-related gene Bax was increased(P<0.01).Conclusions Oleanolic acid can inhibit the growth of human liver cancer HCCLM3 cells transplanted tumors in nude mice,and its mechanism may be related to the inhibition of cell proliferation and the induction of apoptosis.