摘要
目的探讨重组人促红细胞生成素(rh EPO)对人肝癌细胞株(human hepatoma cell line,Hep G2)裸鼠皮下移植瘤的生长影响,及其与促红细胞生成素(EPO)、促红细胞生成素受体(EPOR)、内皮生长因子(VEGF)、Bcl-2(B淋巴细胞/白血病-2基因)表达的时效和量效关系。方法在BALB/C裸鼠皮下接种人肝癌细胞(Hep G2)建立模型,成瘤后随机将其分为rh EPO低剂量组、高剂量组,阴性对照组(PBS组),分组给药,分别于第2周和第4周观察各组肿瘤生长情况,采用HE染色观察肿瘤组织病理情况,应用real-time PCR技术检测肿瘤组织中EPO、EPOR、VEGF、Bcl-2的m RNA水平。结果对照组的肿瘤体积增长幅度均高于实验组肿瘤体积增长幅度;real-time PCR的结果显示:1与对照组比较,第2周高剂量组和低剂量组EPO m RNA表达降低,差异无统计学意义(P>0.05);EPOR m RNA表达降低(P<0.05);VEGF、Bcl-2 m RNA表达均略升高,但差异无统计学意义(P>0.05)。第4周高剂量组和低剂量组EPO、EPOR m RNA较对照组表达降低(P<0.05);EPO高剂量组的VEGF、Bcl-2 m RNA较对照组表达降低(P<0.05),EPO低剂量组的VEGF、Bcl-2 m RNA较对照组表达降低,但差异无统计学意义(P>0.05)。2第2周高剂量组4个基因表达均较低剂量组降低,但差异无统计学意义(P>0.05)。第4周高剂量组EPO、EPOR m RNA表达较低剂量组略有升高,但差异无统计学意义(P>0.05);高剂量组VEGF、Bcl-2 m RNA表达较低剂量组降低,但差异无统计学意义(P>0.05)。3EPO高剂量组第4周与第2周比较4个基因的表达均有所下降,但差异无统计学意义(P>0.05);EPO低剂量组第4周与第2周比较EPO m RNA表达下降(P<0.05),EPOR、VEGF、Bcl-2m RNA表达有所下降,但差异无统计学意义(P>0.05)。结论不同剂量的rh EPO及rh EPO作用时间的不同对Hep G2肿瘤组织的VEGF、Bcl-2 m RNA表达均无明显影响,无直接证据显示rh EPO可引发Hep G2皮下移植瘤的生长。
Objective To investigate the effect of recombinant human erythropoietin(rh EPO) on human hepatoma cell line(Hep G2) grown in nude mice with subcutaneous tumors, and the relationship with erythropoietin(EPO), erythropoietin receptor(EPOR), endothelial growth factor(VEGF), Bcl-2(B lymphocytes/leukemia-2 gene) expression in time-effect and dose-effect. Methods The model in BALB/C mice were inoculated subcutaneously with Hep G2. After tumor formation, the mice were randomly divided into rh EPO low dose group, high dose group and negative control group(PBS group). The mice were administered. The second and the fourth week of each group tumor growth was observed by HE staining of tumor tissue pathology, and applied real-time PCR technology to detect tumors in m RNA levels of EPO, EPOR, VEGF, Bcl-2. Results The tumor volume growth rate in the control group were higher than the experimental group; real-time PCR results showed that:1 Compared with the control group, high-dose group and low dose group EPO m RNA expression in the two weeks group were lower, the differences were not statistically significant(P>0.05); EPOR m RNA expression decreased(P<0.05); VEGF, Bcl-2m RNA expression increased slightly, but the differences were not statistically significant(P >0.05); in the four weeks group,high dose group and low dose group EPO, EPOR m RNA expression were lower than the control group(P<0.05); in high-dose group,VEGF, Bcl-2 m RNA expression were lower than the control group(P<0.05), in low-dose group, VEGF, Bcl-2 m RNA expression were lower than the control group, but the differences were not statistically significant(P>0.05). 2 In the two weeks group, high dose group compared with low-dose group,the four genes decreased, but the differences were not statistically significant(P>0.05). In the four weeks group, high dose group compared with low-dose group EPO, EPOR m RNA expression increased slightly, but the differences were not statistically significant(P>0.05); high dose group compared with low-dose group VEGF, Bcl-2 m RNA expression decreased, but the differences were not statistically significant(P >0.05). 3 In rh EPO high dose group, four weeks group compared with two weeks group, four genes expression decreased slightly, but the differences were not statistically significant(P>0.05); in rh EPO low dose group, four weeks group compared with two weeks group, EPO m RNA expression decreased(P<0.05), EPOR, VEGF, Bcl-2 m RNA expression decreased, but the differences were not statistically significant(P>0.05). Conclusion Different doses of rh EPO and rh EPO different time used on the model of Hep G2,have no significant effect on VEGF,Bcl-2 m RNA expression, no direct evidence shows that rh EPO can influence Hep G2 subcutaneous tumor growth.
出处
《中国现代医药杂志》
2016年第3期21-25,共5页
Modern Medicine Journal of China
