PAX3-FKHR转染对横纹肌肉瘤RD细胞分化及MRF4表达的影响

Influences of PAX3-FKHR Transfection on Expression of MRF4and Differentiation of Cultured Human Rhabdomyosarcoma RD Cells

背景与目的:研究报道融合基因PAX3-FKHR与腺泡型横纹肌肉瘤的发生有着密切的关系,但它对生肌调节因子的作用还不清楚。本研究拟探讨融合基因PAX3-FKHR对胚胎型横纹肌肉瘤细胞分化及MRF4表达的影响。方法:构建PAX3-FKHR的真核表达质粒,采用脂质体法将MRF4、PAX3-FKHR先后转染入人胚胎型横纹肌肉瘤RD细胞,观察转染前后RD细胞生物学特性、形态学改变以及肌分化标志蛋白MHC、α-actin的表达。结果:RD细胞在转染PAX3-FKHR后细胞的生长速度、生长方式及MHC、α-actin的表达均无显著的变化;在表达MRF4的RD细胞中转入PAX3-FKHR后,细胞MRF4表达无明显变化,细胞形态向低分化方向变化,细胞MHC和α-actin表达显著下降。结论:融合基因PAX3-FKHR转染对培养RD细胞的生物学特性及分化无明显影响,并不影响MRF4的表达,但可逆转MRF4促RD细胞分化的作用。

BACKGROUND &OBJECTIVE:There were a few reports about the role of PAX3-FKHR fusion g ene in t he oncog enesis of alveolar rhabdomyosarcoma (ARMS );however,its effect on muscle-prod ucing factor (MPF)is not clear.This study was desig ned to observe the effects of fusion g ene PAX3-FKHR transfection on expressi on of MRF4and differentiation of cul tured human rhabdomyosarcoma RD cells.METHODS :To construct an eukaryotic expression vector of PAX3-FKHR g ene;the plasmids of MRF4cDNA and PAX3-FKHR cDNA were transfected int o cultured RD cells with lipofectin methods.The chang es of cells g rowth,morpholog ic and the expression of M HC andα-actin were observed.RESULTS :There was no sig nificant chang es in R D cells after transfection with PAX3-FKHR g ene in g rowth rate,g rowth manner and the expression of MHC andα-actin.In MRF4expressing RD cells a fter transfected with PAX3-FKHR,althoug h the expression of MRF4was not chan g ed,morpholog ic appearance showed a low differentiation;expression of MHC andα-actin sig nificantly decreased.CONCLUSION:There is no obvious influence on biolog ical character and differe ntiation in RD cells with transfection of PAX3-FKHR g ene;PAX3-FKHR does not i nhibit the expression of MRF4,but ca n reverse the effect of MRF4on promoting the differentiation of RD cell.