VEGF-C在宫颈癌抗凋亡分子机制中的研究  

Effect of VEGF-C Gene Transfection on the Expression of VEGF-C in Human Cervical Carcinoma HeLa Cells and the Molecular Mechnisms of Its Anti-apoptosis Effect

作　　者：陈星[1] 王美芬[1] 吴朝阳[1] 任虹平[1] 郑灵芝[1] 郑曙民[2] 熊冬生[3] 杨纯正[4] 糜若然[3] 

机构地区：[1]温州医学院附属浙江省台州医院妇产科,浙江省临海市317000 [2]山西省肿瘤医院 [3]天津医科大学总医院 [4]中国医学科学院协和医科大学天津血液病学研究所

出　　处：《中国肿瘤临床》2009年第21期1248-1252,共5页Chinese Journal of Clinical Oncology

基　　金：浙江省卫生厅科技基金(编号:2008196B);浙江省台州市科技基金资助(编号:08YK14)

摘　　要：目的:探讨脂质体介导VEGF-C基因转染人宫颈癌HeLa细胞及其对宫颈癌抗凋亡分子机制的研究。方法:前期构建的真核表达载体pcDNA3.1(+)/VEGF-C,用脂质体介导转染人宫颈癌HeLa细胞,并加压筛选获得转染成功的细胞株,经半定量RT-PCR检测转染后VEGF-C表达水平,ELISA检测培养上清中VEGF-C的表达。对转染成功的细胞检测NF-κB、bcl-2基因的表达。结果:在mRNA水平,转染组VEGF-C明显高于空载体组和未转染组;ELISA检测转染组(678.73±38.92ng/mL),也明显高于空载体组(129.52±50.73ng/ml),和未转染组(123.05±55.83ng/mL),成功构建了高表达VEGF-C的宫颈癌细胞株HeLa/S1;在HeLa/S1组NF-κR的表达(2.06±0.09 vs 1.35±0.02 vs 1.38±0.02 P<0.05),bcl-2的表达(2.02±0.67 vs 0.41±0.06 vs 0.37±0.06 P<0.05)明显高于空载体组和未转染组。结论:脂质体介导VEGF-C基因转染人宫颈癌HeLa可显著增加VEGF-C表达,推测高表达的VEGF-C可激活NF-κB,使抗凋亡基因bcl-2高表达,从而促进肿瘤细胞的生长。Objective： To explore the effect of VEGF-C gene transfection on the expression of VEGF-C in human cervical carcinoma HeLa cells and the mechanisms of its anti-apoptosis effect. Methods： The con- structed pcDNA3.1（＋）NEGF-C vector was transformed into human cervical cancer HeLa cells and was selected by G418. The changes in the expression level of VEGF-C mRNA and protein were determined by semi-quantitive RT-PCR and ELISA. HeLa cells with overexpression of VEGF-C were named as HeLa/SI. The expression level of NF-KB and bcl-2 mRNA was determined by RT-PCR in transfected cells. Results： After transfection by liposome, the VEGF-C mRNA level and the expression of VEGF-C protein in transfected cells were higher than those in the control groups. HeLa/S1 cell line was successfully established. In HeLa/S1 cells, the expression of NF-KB （2.06±0.09 vs 1.35±0.02 vs 1.38±0.02 P〈0.05） and bcl-2 gene mRNA （2.02± 0.67 vs 0.41± 0.06 vs 0.37 ± 0.06, P〈0.05） level were higher than those in the control groups. Conclusion： VEGF-C gene transfection by liposome can increase the expression of VEGF-C in human cervical cancer HeLa cells. NF-KB is stimulated and induces the overexpression of bcl-2 gene in HeLa/S1 cells.

关 键 词：血管内皮生长因子-C BCL-2 NF—κB 宫颈癌 

分 类 号：R737.33[医药卫生—肿瘤]