虎杖苷通过调节癌基因表达对小鼠骨肉瘤生长的抑制作用

Inhibitory Effect of Polygonin on the Growth of Osteosarcoma in Mice by Regulating Oncogene Expression and Promoting Cell Apoptosis

目的 探讨虎杖苷通过调节癌基因对小鼠生长骨肉瘤生长的抑制作用。方法 58只小鼠皮下注射U2OS细胞悬液建立骨肉瘤荷瘤小鼠,随机分为模型组、虎杖苷低剂量组、虎杖苷中剂量组、虎杖苷高剂量组和顺铂组。虎杖苷低、中和高剂量组小鼠分别腹腔注射虎杖苷50、100和200 mg/kg;顺铂组小鼠腹腔注射顺铂2 mg/kg, 1次/天,连续5 d;模型组小鼠腹腔注射等量生理盐水。测定肿瘤质量,计算抑瘤率;HE染色观察肿瘤细胞形态;TUNEL染色检测肿瘤细胞凋亡率;qRT-PCR法检测c-Myc、p53和PTEN mRNA水平,蛋白印迹法检测c-Myc、p53、PTEN、Bax和Bcl-2蛋白水平。结果 模型组肿瘤细胞形态正常,核大深染,细胞膜完整;虎杖苷低、中和高剂量组以及顺铂组细胞核染色不同程度变淡,细胞核固缩,细胞密度降低,细胞出现坏死,细胞核出现分裂,其中以顺铂组和虎杖苷高剂量组最为显著。与模型组比较,虎杖苷低、中和高剂量组以及顺铂组肿瘤质量、p53和c-Myc mRNA水平、Bcl-2、p53和c-Myc和蛋白相对表达量降低,且顺铂组<虎杖苷高剂量组<虎杖苷中剂量组<虎杖苷低剂量组;而细胞凋亡率、PTEN mRNA以及Bax和PTEN蛋白相对表达量水平升高(P<0.05)。与虎杖苷低剂量组比较,虎杖苷中剂量组和高剂量组以及顺铂组抑瘤率升高,且顺铂组>虎杖苷高剂量组>虎杖苷中剂量组(P<0.05)。结论 虎杖苷通过促进抑癌基因表达,抑制促癌基因表达,调节细胞凋亡相关蛋白,促进癌细胞凋亡,抑制骨肉瘤生长。

Objective To investigate the inhibitory effect of polygonin on growth of osteosarcoma in mice by regulating oncogenes.Methods 58 mice were injected subcutaneously with U2OS cell suspension to establish osteosarcoma tumor bearing mice,and were randomly divided into model group,cuspidin low-dose group,Cuspidin medium-dose group,Cuspidin high-dose group and cisplatin group.Cuspidin low-dose and high-dose groups were injected intraperitoneally with 50,100 and 200 mg/kg cuspidin,respectively.Mice in cisplatin group were intraperitoneally injected with cisplatin 2 mg/kg once a day for consecutive 5 days,and mice in model group were intraperitoneally injected with the same amount of normal saline.Tumor mass was determined and tumor inhibition rate was calculated.The morphology of tumor cells was observed by HE staining.TUNEL staining was used to detect the apoptosis rate of tumor cells.mRNA levels of c-Myc,p53 and PTEN were detected by qRT-PCR,and protein levels of c-Myc,p53,PTEN,Bax and Bcl-2 were detected by western blotting.Results The tumor cell morphology of model group was normal,the nuclei were large and deeply stained,and the cell membrane was intact.The nuclear staining of cuspidin low-dose,medium-dose and high-dose groups and cisplatin groups was lightest to varying degrees,including nuclear pyretosis,cell density reduction,cell necrosis and cell division,among which the cisplatin group and cuspidin high-dose group were the most significant.Compared with model group,tumor mass,mRNA levels of p53 and c-Myc,Bcl-2,p53 and c-Myc and protein relative expressions of cuspidin low-dose,medium-dose and low-dose cuspidin groups and cisplatin groups decreased,and cisplatin group<cuspidin hig-[LM]h-dose group<cuspidin medium-dose group<cuspidin low-dose group.The apoptosis rate,PTEN mRNA and the relative expression levels of Bax and PTEN protein were increased(P<0.05).Compared with cuspidin low-dose group,the inhibitory rate of cuspidin neutralization high-dose group and cisplatin group was increased,and cisplatin group was>Cuspidin high-dose group>Cuspidin medium-dose group(P<0.05).Conclusion Polygonin can promote the apoptosis of cancer cells and inhibit the growth of osteosarcoma by promoting the expression of tumor suppressor genes,inhibiting the expression of tumor protor genes,regulating apoptosis related proteins.