Xin Xiao-tang, Yin De-zhen, Lan Hai, Chen Cong, Liu Bo, Yang Shang-you
Department of Orthopaedic Surgery, Weihai Municipal Hospital, Weihai, China.
Zhonghua Wai Ke Za Zhi. 2011 Aug 1;49(8):746-51.
To examine the influence of vascular endothelial growth factors (VEGF) in controlling the growth of an experimental osteosarcoma in mice by performing retrovirus-mediated sFlt-1 gene modification.
From March to October 2010 human osteosarcoma G-292 cells were in vitro infected with retroviral vectors encoding soluble Flt-1 or LacZ gene before transplanted into proximal tibiae of immune deficient SCID mice to establish experimental orthotopic osteosarcoma. Daily observation and biweekly microCT were performed to monitor tumor development and progression till sacrifice at 8 weeks after tumor cell inoculation for histological and molecular analyses.
Successful transgene expression was confirmed in the culture media of sFlt-1 transduced G-292 cells using ELISA, and with positive X-gal staining of the LacZ transduced cells. Noteworthy tumors were grown in all mice on the tibiae receiving G-292 cell inoculation, with clear detection on microCT images starting 2 weeks after inoculation. Over the time period, tumors derived from sFlt-1 transduced G-292 cells were distinctively smaller in size compared to the ones from wide-type G-292 and G-292-LacZ cells. Histology showed typical osteosarcoma characteristics including severe cellular pleomorphism, bone erosions, and neo-vascularization. Real-time polymerase chain reaction indicated significantly higher sFlt-1 expression in sFlt-1 transduced groups than the wild-type G-292 or LacZ treated groups. Strong expression of oncogenes c-myc and c-fos were also obvious, along with the expression of VEGF in the primary tumor tissue.
Retrovirus-mediated sFLT-1 gene modification decelerates the osteosarcoma tumor growth in this murine model.
通过进行逆转录病毒介导的sFlt-1基因修饰，研究血管内皮生长因子（VEGF）对小鼠实验性骨肉瘤生长的控制作用。
2010年3月至10月，将人骨肉瘤G-292细胞在体外感染编码可溶性Flt-1或LacZ基因的逆转录病毒载体，然后移植到免疫缺陷SCID小鼠的胫骨近端，建立实验性原位骨肉瘤模型。每天进行观察，每两周进行一次显微CT检查，以监测肿瘤的发生和进展，直至在接种肿瘤细胞8周后处死小鼠进行组织学和分子分析。
使用酶联免疫吸附测定法（ELISA）在sFlt-1转导的G-292细胞的培养基中证实了转基因的成功表达，并且LacZ转导的细胞进行了阳性X-半乳糖苷酶染色。值得注意的是，所有接种G-292细胞的小鼠胫骨上均长出肿瘤，接种后2周在显微CT图像上即可清晰检测到。在此期间，与野生型G-292和G-292-LacZ细胞来源的肿瘤相比，sFlt-1转导的G-292细胞来源的肿瘤体积明显更小。组织学显示典型的骨肉瘤特征，包括严重的细胞多形性、骨质侵蚀和新血管形成。实时聚合酶链反应表明，sFlt-1转导组中的sFlt-1表达明显高于野生型G-292或LacZ处理组。癌基因c-myc和c-fos的强表达也很明显，同时原发性肿瘤组织中也有VEGF的表达。
在该小鼠模型中，逆转录病毒介导的sFLT-1基因修饰可减缓骨肉瘤肿瘤的生长。
