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目的检测EFEMP1在骨肉瘤组织及细胞株F4和F5M2中的表达,探讨EFEMP1在骨肉瘤细胞中的表达,明确其对骨肉瘤细胞侵袭和迁移的影响。方法采用免疫组织化学SP法检测45例临床骨肉瘤及12例骨软骨瘤病例肿瘤组织中EFEMP1表达;采用免疫印迹法(Western blot)及逆转录定量PCR(qRT-PCR)检测具有不同转移潜能的骨肉瘤细胞株(F4、F5M2)中EFEMP1的表达水平;Tanswell实验检测重组人EFEMP1对F5M2细胞侵袭和迁移的影响。结果免疫组化结果显示在45例骨肉瘤标本中,有42例EFEMP1阳性(93.3%),其中强阳性为29例(64.4%),3例(6.7%)为阴性。骨软骨瘤中EFEMP1表达均为阴性。qRT-PCR检测以及Western-blot结果显示,在mRNA和蛋白水平上,骨肉瘤细胞株F4和F5M2中均有EFEMP1的表达,F5M2细胞株EFEMP1的表达高于F4细胞株,Western-blot显示EFEMP1在F4和F5中均有表达,且在F5M2中EFEMP1表达水平高于F4(P<0.05),这一结果与qRT-PCR检测结果一致。Tanswell实验结果显示,与阳性对照组和空白对照组相比,实验组F5M2细胞的侵袭和迁移能力显著增加,重组人全长蛋白EFEMP1可促进骨肉瘤细胞的侵袭和迁移能力。结论EFEMP1在人骨肉瘤组织中表达水平增高,骨肉瘤细胞株F4及F5M2中EFEMP1表达阳性,且高转移骨肉瘤细胞株F5M2中EFEMP1表达量较低转移细胞株F4表达水平明显增高,重组人EFEMP1能够促进骨肉瘤细胞侵袭及转移。EFEMP1可为骨肉瘤患者早期诊断和治疗提供一个新的策略。
Objective To detect the expression of EFEMP1 in osteosarcoma tissues and cell lines F4 and F5M2 and to investigate the expression of EFEMP1 in osteosarcoma cells and to determine its effect on the invasion and migration of osteosarcoma cells. Methods Immunohistochemical SP method was used to detect the expression of EFEMP1 in 45 osteosarcoma cases and 12 osteochondroma cases. The expression of EFEMP1 was detected by Western blot and qRT-PCR The expression of EFEMP1 in osteosarcoma cell lines (F4, F5M2) was detected by Tanswell assay. The effect of recombinant human EFEMP1 on the invasion and migration of F5M2 cells was detected by Tanswell assay. Results Immunohistochemical results showed that 42 of 45 osteosarcoma specimens were positive for EFEMP1 (93.3%), of which 29 (64.4%) were strongly positive and 3 (6.7%) were negative. The expression of EFEMP1 in osteochondroma was negative. qRT-PCR and Western-blot showed that the expression of EFEMP1 in both F4 and F5M2 cells was higher than that of F4 cells in the F5M2 cell line at mRNA and protein levels. Western-blot showed that the expression of EFEMP1 F4 and F5, and the expression of EFEMP1 in F5M2 was higher than that in F4 (P <0.05). This result was consistent with the results of qRT-PCR. Tanswell experimental results showed that compared with the positive control group and the blank control group, the invasion and migration ability of F5M2 cells in the experimental group were significantly increased, and the recombinant human full-length protein EFEMP1 can promote the invasion and migration of the osteosarcoma cells. Conclusion The expression of EFEMP1 in human osteosarcoma tissue is increased, the expression of EFEMP1 in osteosarcoma cell lines F4 and F5M2 is positive, and the expression of EF4 in highly metastatic osteosarcoma cell line F5M2 is significantly higher than that of EFEMP1 Promote osteosarcoma cell invasion and metastasis. EFEMP1 can provide a new strategy for the early diagnosis and treatment of osteosarcoma patients.