Li Guang-Hui, Wang Dong-Lin, Wang Ying, Chen Zheng-Tang, Li De-Zhi
Military Center of Tumor Treatment, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, P. R. China.
Ai Zheng. 2003 Jan;22(1):30-4.
BACKGROUND & OBJECTIVE: There were a few reports about the role of PAX3-FKHR fusion gene in the oncogenesis of alveolar rhabdomyosarcoma (ARMS); however, its effect on muscle-producing factor (MPF) is not clear. This study was designed to observe the effects of fusion gene PAX3-FKHR transfection on expression of MRF4 and differentiation of cultured human rhabdomyosarcoma RD cells.
To construct an eukaryotic expression vector of PAX3-FKHR gene; the plasmids of MRF4 cDNA and PAX3-FKHR cDNA were transfected into cultured RD cells with lipofectin methods. The changes of cells growth, morphologic and the expression of MHC and alpha-actin were observed.
There was no significant changes in RD cells after transfection with PAX3-FKHR gene in growth rate, growth manner and the expression of MHC and alpha-actin. In MRF4 expressing RD cells after transfected with PAX3-FKHR, although the expression of MRF4 was not changed, morphologic appearance showed a low differentiation; expression of MHC and alpha-actin significantly decreased.
There is no obvious influence on biological character and differentiation in RD cells with transfection of PAX3-FKHR gene; PAX3-FKHR does not inhibit the expression of MRF4, but can reverse the effect of MRF4 on promoting the differentiation of RD cell.
关于PAX3-FKHR融合基因在肺泡横纹肌肉瘤(ARMS)发生中的作用已有一些报道;然而,其对肌肉生成因子(MPF)的影响尚不清楚。本研究旨在观察融合基因PAX3-FKHR转染对人横纹肌肉瘤RD细胞MRF4表达及分化的影响。
构建PAX3-FKHR基因的真核表达载体;采用脂质体法将MRF4 cDNA和PAX3-FKHR cDNA质粒转染至培养的RD细胞中。观察细胞生长、形态变化以及MHC和α-肌动蛋白的表达情况。
用PAX3-FKHR基因转染RD细胞后,其生长速率、生长方式以及MHC和α-肌动蛋白的表达均无明显变化。在转染PAX3-FKHR后的MRF4表达阳性的RD细胞中,虽然MRF4的表达未改变,但形态学表现为低分化;MHC和α-肌动蛋白的表达显著降低。
PAX3-FKHR基因转染对RD细胞的生物学特性和分化无明显影响;PAX3-FKHR不抑制MRF4的表达,但可逆转MRF4促进RD细胞分化的作用。