瘦素及胰岛素对人类肝细胞癌细胞系细胞瘦素受体mRNA表达的作用  

Effects of leptin and insulin on leptin receptors mRNA expression in human hepatocellular HepG2 cells

作　　者：刘正娟[1] 姚兴家[1] 王玉川[2] 王德颖[2] 陈竹名[2] 翟玲玲[1] 

机构地区：[1]中国医科大学附属二院儿科,沈阳110004 [2]大连医科大学附属二院儿科

出　　处：《中华医学杂志》2005年第6期396-399,共4页National Medical Journal of China

基　　金：辽宁省自然科学基金资助项目(9910500206)

摘　　要：目的研究瘦素及胰岛素对瘦素受体(OB-R)的调节作用.方法以人类肝细胞癌细胞系(HepG2)为实验摸型,采用6孔板培养皿培养HepG2细胞,4个孔作为一个浓度组,通过DNA序列测定及半定量的逆转录-聚合酶链反应等方法,分别观察了0、10-9、10-8、10-7、10-6 mol/L 浓度的人类瘦素及胰岛素对HepG2细胞内瘦素受体mRNA表达水平的影响.结果通过RT-PCR及DNA序列测定,我们证明了HepG2细胞含有瘦素长型受体(Ob-Rb)及短型受体(OB-Ra:OB-R219.3);通过半定量RT-PCR,我们检测了Ob-Rb 和OB-R219.3的mRNA表达水平.当HepG2细胞与不同浓度的瘦素孵育24 h后,10-7～10-6 mol/L浓度的瘦素明显抑制了OB-Rb的mRNA表达,并在10-6 mol/L浓度时达到最大抑制为43%;同样10-8～10-6mol/L浓度的瘦素亦明显抑制了OB-R219.3的mRNA表达,并在10-6 mol/L浓度时达到最大抑制为49%.当HepG2细胞与不同浓度的胰岛素孵育24 h后, OB-Rb及OB-R219.3的mRNA表达水平无改变.结论在HepG2细胞内瘦素对OB-Rb及OB-R219.3的mRNA表达有下调作用,而胰岛素对OB-Rb及OB-R219.3的mRNA表达无调节作用,这对与我们了解瘦素抵抗及瘦素的外周功能具有重要意义.Objective To study the leptin receptor isoforms regulation by leptin and insulin.Methods Human hepatocellular carcinoma cells of the line HepG2 were cultured in DMEM containing 10% FBS in six well plate and were incubated for 24 hours in serum free medium containing 0,10 -9 ,10 -8 ,10 -7 , and 10 -6 mol/L of human leptin or insulin. Using the semi quantitative RT PCR technique, the mRNA expressions of long (OB Rb) and short (OB Ra:OB R219 3) leptin receptor isoforms were measured. Results OB Rb and OB R219 3 mRNAs were expressed in this cell line. Leptin of the concentrations of 10 -7 ~10 -6 mol/L significantly inhibited the OB Rb mRNA expression, with the maximum decrease (by 43%) at the concentration of 10 -6 mol/L. Similarly the mRNA expression of OB R219 3 was also markedly reduced in cells treated with leptin of the concentrations of 10 -8 ~10 -6 (mol/L), with the maximum inhibition (by 49%) at the concentrations of 10 -6 mol/L. Insulin showed no effect on OB Rb and OB R219 3 mRNAs expression in HepG2 cell.Conclusion In HepG2 cells, leptin down regulates the expressions of OB Rb and OB R219 3 mRNAs, and insulin has no effect on OB Rb and OB R219 3 mRNAs, which contributes at least partly to an understanding of the mechanism of leptin resistance in vivo and suggests that leptin induced receptor down regulation may be relevant to leptin resistance at sites of peripheral action.

关 键 词：MRNA表达 HEPG2细胞 胰岛素 瘦素受体 肝细胞癌 癌细胞系 OB 浓度 Ob-Rb 孵育 

分 类 号：R735.7[医药卫生—肿瘤] R587[医药卫生—临床医学]