不同转移潜能的人体横纹肌肉瘤细胞系细胞间通讯的研究

Studies of intercellular communication in human rhabdomyosarcoma cell lines of different metastatic potential

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 relationship between intercellular communication and three human rhabdomyosarcoma (RMS) cell lines of different metastatic potential. Methods Indirect immunofluorescent staining and laser scanning confocal microscope (ISCM) were used to detect connexin 43 (CX43), a molecule related with gap junctional intercellular communication (GJIC). Fluorescence redistribution after photobleaching(FRAP) was used to detect function of GJIC. Results A high level of CX43 protein expression was revealed in normal human myoblasts. CX43 protein was mainly localized on the surface of cells and sometimes in cytoplasm. CX43 expression was decreased in RMS cells. Both the detection rates and fluorescent intensity of CX43 decreased when the metastatic potential of RMS increased (P<0.05). In contrast to normal, the fluorescence recovery rates of the RMS cells decreased and there was a negative correlation between the function of GJIC and malignancy of RMS cell lines (P<0.05). Conclusion Varying degrees of GJIC inhibition may correlate with different metastatic potential of RMS. This may help to determine the malignant behavior of RMS and be used as a prognostic index of RMS .