中华胃肠外科杂志
中華胃腸外科雜誌
중화위장외과잡지
CHINESE JOURNAL OF GASTROINTESTINAL SURGERY
2009年
4期
416-419
,共4页
郑朝旭%吴荣耀%陈流华%谭敏
鄭朝旭%吳榮耀%陳流華%譚敏
정조욱%오영요%진류화%담민
受体%血管内皮生长因子%转染%结肠肿瘤%Lovo细胞
受體%血管內皮生長因子%轉染%結腸腫瘤%Lovo細胞
수체%혈관내피생장인자%전염%결장종류%Lovo세포
Reeeptors,vascular endothelial growth factor%Transfection%Colonic neoplasms%Lovo cell line
目的 研究人可溶性血管内皮生长肉子受体1(sFlt-1)基因转染结肠癌Lovo细胞后对其细胞生长及培养上清液VEGF含量的影响.方法 使用脂质体介导方法把含sFlt-1基因的重组质粒pcDNA3-sFh-1转染Lovo细胞.通过RT-PCR及ELISA法鉴定sFlt-1的表达,MTT比色法及ELISA法检测该蛋白对Lovo细胞生长及VEGF含鼍的影响.结果 pcDNA3-sFlt-1成功转染Lovo细胞,ELISA法检测到转染后培养上清液中sFlt-1蛋白的表达,MTT实验显示此培养上清液可抑制Lovo细胞的生长,转染后2、14、21和28 d细胞培养上清的抑瘤率分别为(23.92±9.16)%、(13.98±10.21)%、(22.54±11.92)%和(33.43±9.34)%,与未转染组比较,差异均有统计学意义(P<0.05.P<0.01);ELISA证实细胞培养上清中VEGF的含量降低,与未转染组比较,差异亦均有统计学意义(P<0.05,P<0.01).结论 将sFlt-1基因转染至结肠癌Lovo细胞后,可获得具有生物活性的sFlt-1蛋白,抑制癌细胞生长.
目的 研究人可溶性血管內皮生長肉子受體1(sFlt-1)基因轉染結腸癌Lovo細胞後對其細胞生長及培養上清液VEGF含量的影響.方法 使用脂質體介導方法把含sFlt-1基因的重組質粒pcDNA3-sFh-1轉染Lovo細胞.通過RT-PCR及ELISA法鑒定sFlt-1的錶達,MTT比色法及ELISA法檢測該蛋白對Lovo細胞生長及VEGF含鼉的影響.結果 pcDNA3-sFlt-1成功轉染Lovo細胞,ELISA法檢測到轉染後培養上清液中sFlt-1蛋白的錶達,MTT實驗顯示此培養上清液可抑製Lovo細胞的生長,轉染後2、14、21和28 d細胞培養上清的抑瘤率分彆為(23.92±9.16)%、(13.98±10.21)%、(22.54±11.92)%和(33.43±9.34)%,與未轉染組比較,差異均有統計學意義(P<0.05.P<0.01);ELISA證實細胞培養上清中VEGF的含量降低,與未轉染組比較,差異亦均有統計學意義(P<0.05,P<0.01).結論 將sFlt-1基因轉染至結腸癌Lovo細胞後,可穫得具有生物活性的sFlt-1蛋白,抑製癌細胞生長.
목적 연구인가용성혈관내피생장육자수체1(sFlt-1)기인전염결장암Lovo세포후대기세포생장급배양상청액VEGF함량적영향.방법 사용지질체개도방법파함sFlt-1기인적중조질립pcDNA3-sFh-1전염Lovo세포.통과RT-PCR급ELISA법감정sFlt-1적표체,MTT비색법급ELISA법검측해단백대Lovo세포생장급VEGF함타적영향.결과 pcDNA3-sFlt-1성공전염Lovo세포,ELISA법검측도전염후배양상청액중sFlt-1단백적표체,MTT실험현시차배양상청액가억제Lovo세포적생장,전염후2、14、21화28 d세포배양상청적억류솔분별위(23.92±9.16)%、(13.98±10.21)%、(22.54±11.92)%화(33.43±9.34)%,여미전염조비교,차이균유통계학의의(P<0.05.P<0.01);ELISA증실세포배양상청중VEGF적함량강저,여미전염조비교,차이역균유통계학의의(P<0.05,P<0.01).결론 장sFlt-1기인전염지결장암Lovo세포후,가획득구유생물활성적sFlt-1단백,억제암세포생장.
Objective To investigate the effect of transfection with human soluble vascular endothelial growth factor receptor-1 (sFlt-1) gene on cell growth and vascular endothelial growth factor (VEGF) concentration of the culture supematant in human colon cancer cell line Lovo. Methods The recombinant plasmid peDNA3-sFlt-1 containing sFlt-1 gene was transfected into Lovo cells by Lipofeetamine 2000, which was identified by RT-PCR and ELISA. The effect of sFlt-1 protein on cell growth and VEGF expression in Lovo cells were investigated by MTT and ELISA. Results The recombinant plasmid peDNA3-sFlt-1 was successfully transfected into Lovo cells. The sFh-1 expression was identified by RT-PCR and ELISA, which inhibited the growth of Lovo cells and reduced the VEGF concentration in the culture supematant compared with control. The inhibitory rates of proliferation of Lovo cells via MTT assay after 2,14,21 and 28 days were (23.92±9.16)%, (13.98±10.21)%,(22.54±11.92)% and (33.43±9.34)% respectively. Compared with the control groups,the differences were significant(P<0.05 ,P<0.01). Conclusion Transfeetion with sFlt-1 gene into Lovo cells results in the expression of sFh-1 protein, which possesses high biological activity and inhibits the growth of cancer cells.
