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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 investigate the liposome-mediated VEGF-C gene transfected HeLa cells and human cervical cancer anti-apoptotic molecular mechanism. Methods: The eukaryotic expression vector pcDNA3.1 (+) / VEGF-C was constructed and transfected into HeLa cells by lipofectamine 2000. HeLa cells were transfected successfully by pressure screening. The expression of VEGF-C after transfection was detected by PCR, and the expression of VEGF-C in culture supernatant was detected by ELISA. The successful transfected cells were detected NF-κB, bcl-2 gene expression. Results: At mRNA level, VEGF-C in transfection group was significantly higher than that in empty vector group and untransfected group (678.73 ± 38.92ng / mL), which was also significantly higher than that in empty vector group (129.52 ± 50.73ng / HepG2 / HepG2 / HepG2 / HepG2 / HepG2 / HepG2 / HepG2 / HepG2 / HepG2 / HepG2 / HepG2 / 1.35 ± 0.02 vs 1.38 ± 0.02 P <0.05). The expression of bcl-2 (2.02 ± 0.67 vs 0.41 ± 0.06 vs 0.37 ± 0.06, P <0.05) was significantly higher than that in empty vector group and non-transfected group. CONCLUSION: Liposome-mediated VEGF-C gene transfected into human cervical cancer HeLa can significantly increase the expression of VEGF-C, speculated that high expression of VEGF-C can activate NF-κB, so that anti-apoptotic gene bcl-2 expression, thus Promote the growth of tumor cells.