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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 & AIM: PAX3-FKHR fusion gene is closely related to the occurrence of acinar rhabdomyosarcoma. However, the role of PAX3-FKHR in regulating myogenic factor remains unclear. This study was to investigate the effect of fusion gene PAX3-FKHR on embryonic rhabdomyosarcoma cell differentiation and MRF4 expression. Methods: The eukaryotic expression plasmids of PAX3-FKHR were constructed. The MRF4 and PAX3-FKHR were transfected into human embryonic rhabdomyosarcoma RD cells by liposome method. The biological characteristics, morphological changes, Differentiation markers MHC, α-actin expression. Results: There was no significant change in the growth rate, growth pattern and the expression of MHC and α-actin in RD cells after PAX3-FKHR transfection. The expression of MRF4 in PAF3-FKHR cells after transfected with RDF cells expressing MRF4 Significant changes in cell morphology to the low differentiation direction, the expression of MHC and α-actin decreased significantly. CONCLUSION: Transfection of fusion gene PAX3-FKHR has no significant effect on the biological characteristics and differentiation of cultured RD cells, does not affect the expression of MRF4, but reverses the effect of MRF4 on differentiation of RD cells.