反义转化生长因子-β1基因逆转骨肉瘤细胞恶性表型的研究

Reverse malignant phenotypes of osteosarcoma cells transfected by antisense TGF-β1 gene

目的 观察反义转化生长因子 (TGF) β1基因对骨肉瘤细胞恶性表型的逆转作用。方法 将反义TGF β1基因转染骨肉瘤细胞MG 63后 ,以G418(4 0 0mg/L)筛选 14d ,获得转染细胞系。分别检测细胞周期、细胞凋亡和明胶酶A(MMP 2 )表达 ,并观察转染细胞软琼脂克隆形成数和裸鼠体内致瘤能力。结果 与MG 63和空载体转染细胞MG pcDNA3相比 ,反义基因转染细胞MG TGF β1(-)的G0 /G1期细胞从 5 6.2 %和 60 .1%增至 71.6% ,S期细胞从 19.1%和 17.8%降至 12 .9% ,MMP 2mRNA表达明显减少。MG TGF β1( )的软琼脂克隆形成数从 48.6± 8.1和45 .2± 4.7减至 2 8.2± 5 .6,裸鼠体内形成肿瘤的体积 (83 .4± 2 7.4)mm3 也明显小于MG 63组(191.3± 2 1.5 )mm3 和MG pcDNA3组 (2 0 4.2± 3 0 .7)mm3 。结论 反义TGF β1基因转染骨肉瘤细胞可以逆转肿瘤细胞的恶性表型 ,抑制肿瘤的侵袭和转移。

Objective To investigate the reversing effects of antisense TGF-β1 gene on the malignant phenotypes of osteosarcoma cells.Methods MG-63 cells were transfected by antisense TGF-β1 gene and selected in DMEM containing G418 (400 mg/L) for 14 days.The effects of antisense TGF-β1 gene on the malignant phenotypes of MG-63 cells were examined by using flow cytometric analysis,RT-PCR,soft sugar test and nude mice inoculation test.Results Compared to MG-63 and MG-pcDNA3,the number of cells in the G 0/G 1 phase of MG-TGF-β1(-) was increased from 56.2% and 60.1% to 71.6%,but that of cells in the S phase was decreased.The clony formation rate of MG-TGF-β1(-) was decreased from 48.6±8.1 and 45.2±4.7 to 28.2±5.6,and the tumorigenicity in nude mice was also decreased.Conclusion The antisense TGFβ1 gene could reverse the malignant phenotype of osteosarcoma cells to some extent and inhibit tumor invasion and metastasis.