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机构地区:[1]大连医科大学附属第二医院普外科,辽宁大连116027
出 处:《实用肿瘤杂志》2005年第1期21-24,共4页Journal of Practical Oncology
摘 要:目的 探讨 VEGF- C反义寡核苷酸 (antisense oligodeoxynucleotide,ASODN )对人乳腺癌细胞 MDA-MB- 4 35黏附、侵袭能力的影响。方法 以 VEGF- C基因编码区为靶点合成反义寡核苷酸 ,脂质体介导转染。 RT-PCR法检测转染后 VEGF- C m RNA表达水平 ;MTT法检测乳腺癌细胞与细胞外基质黏附 ;利用 Transwell小室 ,检测乳腺癌细胞侵袭人工基底膜能力 ;Western- blot法检测乳腺癌细胞中 MMP- 2蛋白表达。结果 与空白对照及正义序列 (sense oligodeoxynucleotide,SODN)转染组比较 ,ASODN组 VEGF- C m RNA水平下调 ,差异有显著性 (t=5 .6 4 ,5 .4 8,P<0 .0 1) ;对胞外基质黏附率明显降低 (χ2 =7.15 ,7.2 9,P<0 .0 5 ) ,穿透重组基底膜数目显著下降 (t=5 .2 9,5 .17,P<0 .0 1) ;细胞裂解液中 MMP- 2蛋白表达显著下调 (t=7.36 ,7.4 3,P<0 .0 1)。结论 VEGF- C基因 ASODN脂质体介导转染可明显抑制乳腺癌细胞体外实验中的黏附和侵袭能力 ;下调 MMP- 2的表达可能是其作用途径。Objective To investigate the effects of V EGF-C antisense oligodeoxynucleotide(ASODN) on the adhesion and invassion of br east cancer cell line MDA-MB-435 in vitro.Methods The ASO DN sequence targeting the coding region of VEGF-C was synthesized and introduce d into breast cancer cells by liposomes.RT-PCR was used to detect the mRNA leve l of VEGF-C;MTT method was used to detect the adhesion of cancer cells to extra cellular matrix. Trans-well chamber was used to detect the invasive capacity of cancer cells to the recombinant basement membrane.Western-blot was used to det ect the expression of MMP-2.Results In ASODN group,the VEGF-C mRNA level was significantly decreased (t=5.64,5.48,P<0.01);the adhesive r ate and the invasive cell number were decreased markedly (χ2=7.15,7.29,P<0. 05/t=4.29,4.17,P<0.01) compared with the control and sense oligodeoxynucle otide (SODN) group. The result of Western-blot showed a down-regulated MMP-2 expression (t=7.36,7.43,P<0.01).Conclusion The ASODN of VEG F-C can inhibit the adhesive and invasive capacity of breast cancer cells in vitro; Down-regulating the expression of MMP-2 may be the mechanism.\;
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