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通过 siRNA 干扰抑制血管内皮生长因子调节人鼻咽癌细胞的生物学行为。

Suppression of vascular endothelial growth factor via siRNA interference modulates the biological behavior of human nasopharyngeal carcinoma cells.

机构信息

Department of Radiation and Medical Oncology, Second Hospital of Yichang, 4 Tiyuchang Road, Yichang, Hubei 443000, China.

出版信息

Jpn J Radiol. 2011 Nov;29(9):615-22. doi: 10.1007/s11604-011-0603-9. Epub 2011 Sep 29.

Abstract

PURPOSE

The aim was to study the effect of vascular endothelial growth factor (VEGF) down-regulation by small interfering (si)RNA-mediated interference (RNAi) on the biological features of nasopharyngeal carcinoma cell line CNE-2.

MATERIALS AND METHODS

The combined plasmids pU-siVEGF and pU-siCONT were transfected into CNE-2 cells with lipofectamine. The transfected cells were placed in fresh medium containing G418. Expression of VEGF mRNA and protein were measured by reverse transcriptase-polymerase chain reaction and Western blot, respectively. The transwell chamber model was employed to test the ability of cell invasion in vitro. The distribution of cell cycle phases was determined by flow cytometry. Cell survival was assessed by clonogenic assays.

RESULTS

Both VEGF mRNA and protein expression were significantly decreased in the pU-siVEGF group compared with controls (P < 0.05). The cell cycle was arrested in the G(1) phase (P < 0.05). A higher apoptotic ratio and lower invasion ability were seen in the pU-siVEGF group. The D(0) (mean lethal dose) and SF(2) values were significantly lower than those in the control group (P < 0.05).

CONCLUSION

Delivery of siRNA targeting VEGF seems efficient in down-regulating VEGF expression and diminishing the growth, proliferation, and invasiveness of CNE-2 cells. It also enhanced the sensitivity of CNE-2 cells to radiation.

摘要

目的

研究通过小干扰 RNA(siRNA)介导的干扰下调血管内皮生长因子(VEGF)对鼻咽癌细胞系 CNE-2 的生物学特性的影响。

材料与方法

采用脂质体将联合质粒 pU-siVEGF 和 pU-siCONT 转染至 CNE-2 细胞中,转染后的细胞置于含有 G418 的新鲜培养基中。采用逆转录-聚合酶链反应和 Western blot 分别检测 VEGF mRNA 和蛋白的表达。采用 Transwell 小室模型检测体外细胞侵袭能力。采用流式细胞术检测细胞周期分布。采用集落形成实验评估细胞存活能力。

结果

与对照组相比,pU-siVEGF 组 VEGF mRNA 和蛋白表达均显著降低(P<0.05)。细胞周期被阻滞在 G1 期(P<0.05)。pU-siVEGF 组细胞凋亡比例更高,侵袭能力更低。pU-siVEGF 组的 D0(平均致死剂量)和 SF2 值均显著低于对照组(P<0.05)。

结论

针对 VEGF 的 siRNA 转染在下调 VEGF 表达、抑制 CNE-2 细胞生长、增殖和侵袭方面具有显著效果,同时增强了 CNE-2 细胞对辐射的敏感性。

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