Yu Yong-Jiang, Li Yu-Min, Hou Xu-Dong, Guo Chao, Cao Nong, Jiao Zuo-Yi
Department of General Surgery, First Hospital of Lanzhou Univercity, Lanzhou, China.
Asian Pac J Cancer Prev. 2012;13(5):1845-9. doi: 10.7314/apjcp.2012.13.5.1845.
Tissue factor (TF) is expressed abnormally in certain types of tumor cells, closely related to invasion and metastasis. The aim of this study was to construct a human gastric cancer cell line SGC7901 stably-transfected with human TF, and observe effects on oxaliplatin-dependent inhibition of invasion and the apoptosis induction.
The target gene TF was obtained from human placenta by nested PCR and introduced into the human gastric cell line SGC7901 through transfection mediated by lipofectamine. Stably-transfected cells were screened using G418. Examples successfully transfected with TF-pcDNA3 recombinant (experimental group), and empty vector pcDNA3 (control group) were incubated with oxaliplatin. Transwell chambers were used to show change in invasive ability. Caspase-3 activity was detected using a colorimetric method and annexin-V/PI double- staining was applied to detect apoptosis.
We generated the human gastric cancer cell line SGC7901/TF successfully, expressing TF stably and efficiently. Compared with the control group, invasion increased, whereas caspase-3 activity and apoptosis rate were decreased in the experimental group.
TF can enhance the invasive capacity of gastric cancer cells in vitro. Its increased expression may reduce invasion inhibition and apoptosis-inducing effects of oxaliplatin and therefore may warrant targeting for improved chemotherapy.
组织因子(TF)在某些类型的肿瘤细胞中异常表达,与侵袭和转移密切相关。本研究旨在构建稳定转染人TF的人胃癌细胞系SGC7901,并观察其对奥沙利铂依赖性侵袭抑制和凋亡诱导的影响。
通过巢式PCR从人胎盘中获得靶基因TF,并通过脂质体介导的转染将其导入人胃癌细胞系SGC7901。使用G418筛选稳定转染的细胞。将成功转染TF-pcDNA3重组体(实验组)和空载体pcDNA3(对照组)的样本与奥沙利铂一起孵育。使用Transwell小室显示侵袭能力的变化。采用比色法检测Caspase-3活性,并应用膜联蛋白V/碘化丙啶双染法检测凋亡。
我们成功构建了人胃癌细胞系SGC7901/TF,其稳定且高效地表达TF。与对照组相比,实验组的侵袭增加,而Caspase-3活性和凋亡率降低。
TF可增强胃癌细胞的体外侵袭能力。其表达增加可能会降低奥沙利铂的侵袭抑制和凋亡诱导作用,因此可能需要针对其进行靶向治疗以改善化疗效果。