Liu Shuyong, Chen Zhiping
Department of Hand and Foot Surgery, Jining No. 1 People's Hospital, Jining, Shandong, China (mainland).
Department of Pediatrics, Jining No. 1 People's Hospital, Jining, Shandong, China (mainland).
Med Sci Monit. 2015 Jul 8;21:1976-82. doi: 10.12659/MSM.893430.
As the most common primary bone tumor, osteosarcoma has an improved survival rates with advancement of treatment methods. A higher rate of metastasis, however, leads to the aggravation of the disease. Studies have shown that some genes, namely osteosarcoma metastasis-related genes, participate in the process of tumor metastasis. The peripheral myelin protein 22 (PMP22) gene has recently been found to be abundantly expressed in the oncogenesis of osteosarcoma. Its detailed role and function in the tumor metastasis, however, remains unknown.
The recombinant retroviral plasmid pcDNA3.1-PMP22 was constructed and used to transfect osteosarcoma cells SOSP-M, whose cell proliferation was measured by MTT method. The formation of tumor cell colony, the cell migration and invasion were also measured. The signal transduction pathway MAPK was further analyzed by Western blotting.
The pcDNA3.1-PMP22 plasmid was confirmed to have a 305bp PMP22 fragment by EcoRI-XhoI dual digestion. Compared to the control group, osteosarcoma cell invasion was significantly facilitated by the transfection of pcDNA3.1-PMP22 plasmid (p<0.05). The recombinant plasmid also significantly potentiated the formation of tumor cell colony and increased the migration and invasion ability of tumor cells (p<0.05 in all cases). Phosphorylated p-ERK and p-P38 were also up-regulated by vector transfection (p<0.05).
Osteosarcoma metastasis-related gene PMP22 participates in the proliferation, invasion, migration and colony formation of osteosarcoma cells possibly via the MAPK signal transduction pathway, providing evidences for further investigation of metastatic mechanism of osteosarcoma.
骨肉瘤作为最常见的原发性骨肿瘤，随着治疗方法的进步，其生存率有所提高。然而，较高的转移率会导致病情加重。研究表明，一些基因，即骨肉瘤转移相关基因，参与肿瘤转移过程。外周髓鞘蛋白22（PMP22）基因最近被发现在骨肉瘤的发生过程中大量表达。然而，其在肿瘤转移中的具体作用和功能仍不清楚。
构建重组逆转录病毒质粒pcDNA3.1-PMP22，并用于转染骨肉瘤细胞SOSP-M，通过MTT法检测其细胞增殖情况。同时检测肿瘤细胞集落的形成、细胞迁移和侵袭情况。通过蛋白质免疫印迹法进一步分析丝裂原活化蛋白激酶（MAPK）信号转导通路。
经EcoRI-XhoI双酶切证实pcDNA3.1-PMP22质粒含有305bp的PMP22片段。与对照组相比，转染pcDNA3.1-PMP22质粒可显著促进骨肉瘤细胞的侵袭（p<0.05）。该重组质粒还显著增强了肿瘤细胞集落的形成，并提高了肿瘤细胞的迁移和侵袭能力（所有情况均p<0.05）。载体转染还上调了磷酸化的p-ERK和p-P38（p<0.05）。
骨肉瘤转移相关基因PMP22可能通过MAPK信号转导通路参与骨肉瘤细胞的增殖、侵袭、迁移和集落形成，为进一步研究骨肉瘤的转移机制提供了依据。
