摘要
目的探讨甲基强的松龙(MP)对肝癌HepG2细胞生长的影响及相关机制。方法MTT法测定不同浓度MP对HepG2细胞增殖的影响;细胞分为6个处理组:对照组、MPⅠ组(10ug/L)、MPⅡ组(20 ug/L)、MPIII组(50 ug/L)、MPIV组(500 ug/L)、MPV组(5000 ug/L);流式细胞仪测定MP对HepG2细胞周期和凋亡情况的作用;荧光定量PCR法测定MP对HepG2细胞VEGF mRNA表达的影响;ELISA法测定细胞培养上清液VEGF浓度。结果50～5000 ug/L的MP作用72 h,能促进HepG2细胞的增殖;MP对HepG2细胞周期和凋亡无影响;不同浓度的MP作用72 h后,HepG2细胞VEGF mRNA的转录和蛋白表达有显著性差异(P<0.01),MP在500 ug/L和5000 ug/L浓度时,VEGF mRNA表达量分别为(7.293±0.409)拷贝数/ul对数值和(8.231±0.216)拷贝数/uL对数值,高于对照组(4.761±0.624)拷贝数/uL对数值,结果有显著性差异(P<0.01),VEGF浓度分别为(11.153±1.552)ug/L和(11.456±1.517)ug/L,高于对照组(7.362±0.510)ug/L,结果有显著性差异(P<0.01)。结论高浓度的MP在体外能促进肝癌HepG2细胞增殖,上调其VEGF mRNA的转录和翻译。
Objective To investigate the efficacy and the possible mechanism of HepG2 cell line induced by methylyprednisolone （MP） in vitro. Methods The proliferation of HepG2 cells treated with MP was determined by MTT assay. Flow cytometry was used to detect cell cycle and apoptosis induced by MP. HepG2 ceils were randomly divided into 6 treatment groups： the control group, the MP group Ⅰ （MP 10 u/L）, group Ⅱ （MP20 ug/L）, group Ⅲ （MP50 ug/L）, group Ⅳ （MPS00 ug/L）, group Ⅴ （MP5000 ug/L）. The expression of VEGF mRNA of HepG2 cells was detected by fluorescent quantitation ploymerase chain reaction （FQ-PCR）. The VEGF concentration of supemate of HepG2 cells was measured by ELISA. Results MP enhanced the proliferation of HepG2 cells at the concentration of 50-5000 ug/L in vitro. The expression of VEGF mRNA was promoted significantly by MP at the concentration of 500-5000 ug/L. The expression of VEGF mRNA in the control group, the MP group Ⅰ , group Ⅱ , group Ⅲ, group Ⅳ, group Ⅴ was （4.761±0.624）, （4.679±0.875）, （5.554±0.541 ）, （5.710±1.22）, （7.293±0.409）, （8.231±0.216） （P〈0.01 ）.The supernate concentration of VEGF was promoted significantly by MP at the concentration of 500-5000 ug/L （P〈0.01）. Conclusion High dose of methylyprednisolone can enhance the growth of hepatocellular carcinoma cell lines HepG2 in vitro and up-regulate the transcription and translation of VEGF mRNA.
出处
《岭南现代临床外科》
2012年第1期16-19,共4页
Lingnan Modern Clinics in Surgery
基金
广东省医学科研基金项目(A2009545)
广州市卫生局基金资助项目(2008-YB-073)
关键词
甲基强的松龙
肝细胞癌
血管内皮生长因子
Methylyprednisolone
Hepatocellular carcinoma
Vascular endothelial growth factor
