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目的 研究细胞间通讯功能在 3种不同转移潜能的人体横纹肌肉瘤细胞系间及其与培养的正常人肌母细胞的差别及其意义。方法 采用间接免疫荧光法 ,用激光扫描共聚焦显微镜定量测定与细胞间通讯有关的间隙连接蛋白 (connexin43,CX43)的含量 ,并用荧光漂白后恢复技术检测间隙连接介导的细胞间通讯的功能。结果 CX43在正常肌母细胞中呈高表达 ,主要定位于细胞膜 ,有时定位于胞质 ,而在横纹肌肉瘤中表达下降 ;CX43阳性率及荧光强度均随着横纹肌肉瘤转移潜能的增加而降低 (P <0 0 5 ) ;荧光漂白后恢复技术结果显示横纹肌肉瘤细胞荧光漂白后恢复率下降而且与横纹肌肉瘤细胞的转移潜能呈负相关 (P <0 0 5 )。结论 不同程度细胞间通讯功能的抑制可能与横纹肌肉瘤的转移潜能不同有关 ,其有可能成为判断横纹肌肉瘤恶性程度及预后的指标之一。
Objective To investigate the differences and significances of intercellular communication between human rhabdomyosarcoma cell lines with different metastatic potentials and their normal human myoblasts. Methods The indirect immunofluorescence method was used to quantitatively measure the content of connexin43 (CX43) associated with cell-cell communication by laser scanning confocal microscopy, and the function of gap junction-mediated cell-cell communication was detected by fluorescence bleaching recovery technique. Results The high expression of CX43 in normal myoblasts was mainly localized in the cell membrane, sometimes located in the cytoplasm and decreased in rhabdomyosarcoma; the positive rate of CX43 and fluorescence intensity decreased with the increase of the metastatic potential of rhabdomyosarcoma (P). <0 0 5 ) ; Recovery results after fluorescence bleaching showed that the recovery rate of rhabdomyosarcoma cells after fluorescent bleaching was negatively correlated with the metastatic potential of rhabdomyosarcoma cells (P < 0.05). Conclusion The inhibition of different levels of intercellular communication may be related to the different metastatic potential of rhabdomyosarcoma, which may be one of the indicators to judge the malignancy and prognosis of rhabdomyosarcoma.