CAJ | 학술논문

目的：检测在人骨肉瘤组织、骨肉瘤MG63细胞及其干细胞中Hippo信号分子TAZ的表达情况，比较骨肉瘤细胞和骨肉瘤干细胞中 TAZ 的表达差异，并探讨其可能的临床意义。方法2010年1月至2012年1月在我院治疗的骨肉瘤患者中，选取原发骨肉瘤组织标本12例，复发的骨肉瘤标本6例，另选6例骨纤维结构不良组织作阴性对照，并贴壁培养骨肉瘤MG63细胞，采用免疫组化方法检测骨肉瘤组织和骨肉瘤MG63细胞中 TAZ 的表达情况；用无血清悬浮培养骨肉瘤 MG63细胞，分离并收集骨肉瘤细胞球，通过定量反转录-聚合酶链反应( RT-PCR )法检测骨肉瘤MG63细胞及骨肉瘤细胞球中胚胎干细胞标志基因( Nanog， Oct4)和 Hippo 信号分子 TAZ 的表达，Western 印记法检测 MG63细胞和细胞球中 TAZ 蛋白的表达情况。结果在12例人骨肉瘤原发组织标本中，3例骨肉瘤组织TAZ表达阳性，而6例骨肉瘤复发组织中TAZ表达全部阳性，在骨肉瘤细胞MG63中TAZ分子表达也呈阳性，而骨纤维结果不良中全部表达阴性。与骨肉瘤MG63细胞相比，RT-PCR显示培养的骨肉瘤细胞球中干细胞标志基因Nanog，Oct4和Hippo信号分子TAZ明显高表达。Western blot 提示细胞球中 TAZ 蛋白表达明显高于骨肉瘤 MG63细胞。结论骨肉瘤组织中存在TAZ分子的表达，在复发骨肉瘤组织中呈高表达；在骨肉瘤细胞株中也存在TAZ分子的表达，而在干细胞中TAZ分子表达更高；提示其与骨肉瘤干细胞的特征具有一定的相关性。
목적：검측재인골육류조직、골육류MG63세포급기간세포중Hippo신호분자TAZ적표체정황，비교골육류세포화골육류간세포중 TAZ 적표체차이，병탐토기가능적림상의의。방법2010년1월지2012년1월재아원치료적골육류환자중，선취원발골육류조직표본12례，복발적골육류표본6례，령선6례골섬유결구불량조직작음성대조，병첩벽배양골육류MG63세포，채용면역조화방법검측골육류조직화골육류MG63세포중 TAZ 적표체정황；용무혈청현부배양골육류 MG63세포，분리병수집골육류세포구，통과정량반전록-취합매련반응( RT-PCR )법검측골육류MG63세포급골육류세포구중배태간세포표지기인( Nanog， Oct4)화 Hippo 신호분자 TAZ 적표체，Western 인기법검측 MG63세포화세포구중 TAZ 단백적표체정황。결과재12례인골육류원발조직표본중，3례골육류조직TAZ표체양성，이6례골육류복발조직중TAZ표체전부양성，재골육류세포MG63중TAZ분자표체야정양성，이골섬유결과불량중전부표체음성。여골육류MG63세포상비，RT-PCR현시배양적골육류세포구중간세포표지기인Nanog，Oct4화Hippo신호분자TAZ명현고표체。Western blot 제시세포구중 TAZ 단백표체명현고우골육류 MG63세포。결론골육류조직중존재TAZ분자적표체，재복발골육류조직중정고표체；재골육류세포주중야존재TAZ분자적표체，이재간세포중TAZ분자표체경고；제시기여골육류간세포적특정구유일정적상관성。
Objective To detect the expressions of transcriptional coactivator with plasma dissociated zircon ( PDZ )-binding motif ( TAZ ) molecules of Hippo signaling pathway in human osteosarcoma tissues, osteosarcoma MG63 cells and stem cells, to compare the expression differences of TAZ molecules in osteosarcoma cells and osteosarcoma stem cells, and to investigate possible clinical signiifcance. Methods 12 primary osteosarcoma tissue specimens and 6 recurrent osteosarcoma specimens were selected from the patients with osteosarcomas treated in our hospital from January 2010 to January 2012, with another 6 patients with ifbrous dysplasia of bone as the negative control group. Osteosarcoma MG63 cells were cultured by adherent method. Immunohistochemical method was used to detect the expressions of TAZ molecules in osteosarcoma tissues and osteosarcoma MG63 cells. Osteosarcoma MG63 cells were cultured in suspension in serum-free medium to separate and collect osteosarcoma spheroids. The expressions of marker genes of embryonic stem cells ( Nanog and Oct4 ) and TAZ molecules of Hippo signaling pathway were detected in osteosarcoma MG63 cells and osteosarcoma spheroids by quantitative reverse transcription-polymerase chain reaction ( RT-PCR ). The expressions of TAZ proteins in MG63 cells and spheroids were detected by western blot. Results The expressions of TAZ molecules were positive in 3 of 12 primary human osteosarcoma tissue specimens, which were also positive in all the recurrent osteosarcoma specimens. The expressions of TAZ molecules were positive in osteosarcoma MG63 cells, and negative expressions of TAZ molecules were found in the control group. Compared with osteosarcoma MG63 cells, RT-PCR analysis showed the expressions were signiifcantly higher in marker genes of stem cells ( Nanog and Oct4 ) in cultured osteosarcoma spheroids and TAZ molecules of Hippo signaling pathway. Based on western blot results, the expressions of TAZ proteins in spheroids were significantly higher than that in osteosarcoma MG63 cells. Conclusions There are TAZ molecules expressed in osteosarcoma tissues, but they are highly expressed in recurrent osteosarcoma tissues. The expressions of TAZ molecules are also found in osteosarcoma cell lines, but they are higher in stem cells. Therefore, it is suggested that TAZ molecules of Hippo signaling pathway may have a certain correlation with the characteristics of osteosarcoma stem cells.