摘要
目的:探讨人cathepsin B(CatB)反义RNA在乳腺癌细胞侵袭与迁移中的作用。方法:构建携CatB反义RNA的重组质粒pBudCE4.1-antiCatB,采用脂质体法将重组质粒瞬时转染人乳腺癌细胞MDA-MB-231。Western blotting法检测MDA-MB-231细胞中CatB蛋白的表达,MTT法检测MDA-MB-231细胞的增殖,细胞-基质黏附实验检测反义CatB对MDA-MB-231细胞黏附能力的影响,体外侵袭、迁移实验分析反义CatB表达对MDA-MB-231细胞体外侵袭和迁移能力的影响。结果:成功构建pBudCE4.1-antiCatB表达载体。转染MDA-MB-231细胞后,与未转染组细胞和转染空载体组细胞相比,转染组MDA-MB-231细胞的CatB蛋白表达水平明显降低[(0.96±0.02)vs(1.98±0.23),(1.84±0.08),P<0.05];转染组细胞的增殖受到明显抑制[(0.255±0.017)vs(0.458±0.033),(0.421±0.022),P<0.01];转染组细胞的黏附(基质胶或纤维黏连蛋白)能力明显下降[(0.054±0.017)vs(0.111±0.018),(0.107±0.017),P<0.01;或(0.052±0.008)vs(0.120±0.014),(0.113±0.009),P<0.01];转染组细胞的侵袭和迁移能力也明显降低[(52.80±7.76)vs(124.00±44.54),(116.80±32.87),P<0.01;(60.25±8.73)vs(132.50±12.15),(119.20±25.13),P<0.01]。结论:CatB反义RNA可抑制乳腺癌细胞体外生长、黏附、迁移和侵袭能力。
Objective ：To investigate the effect of antisense cathepsin B （CatB） RNA on the invasion and migration of human breast carcinoma cells. Methods： PBudCE4. 1-antiCatB recombinant plasmid carrying antisense eathepsin B （CatB） gene was constructed and transfeeted into breast carcinoma cell line MDA-MB-231 by lipofection system. The expression of CatB protein in MDA-MB-231 cells was detected by Western blotting analysis; the proliferation of MDA-MB- 231 cells was determined by MTT assay. Cell-matrix adhesion assay was used to examine the effect of anti-CatB on adhesion ability of MDA-MB-231 cells. The effects of anti-CatB on the invasion and migration abilities of MDA-MB-231 cells were measured by invasion and migration transwell system. Results： The recombinant plasmid PBudCE4. 1-antiCatB was successfully constructed. Expression of CatB protein in MDA-MB-231 cells was decreased after PBudCE4. 1-antiCatB transfection compared with those in untransfected and mock-vehicle transfeeted cells（ [0.96 ±0.02 ] vs [ 1.98 ± 0. 23 ], [ 1.84 ± 0. 08 ], P 〈 0.05 ） ; the proliferation of MDA-MB-231 cells was also inhibited in PBudCE4. 1-antiCatB transfected group （ [ 0. 255 ± 0.017 ] vs [ 0.458 ± 0.033 ], [ 0. 421 ± 0. 022 ], P 〈 0. 01 ） ; and the adhesion abilities （ binding to matrix or fibronectin） were decreased（ [0.054 ±0.017] vs [0. 111 ±0.018], [0. 107 ±0.017], P〈0. 01 ; [0.052 ± 0.008 ] vs [ 0.120 ± 0.014 ], [ 0. 113 ± 0.009 ], P 〈 0.01 ）. Transwell asaay showed that the invasion and migration a-bilities were inhibited in PBudCFA. 1-antiCatB transfected group compared with those in the non-transfection and mock-vehicle transfected groups（ [ 52.80 ± 7.76 ] vs [ 124.00 ± 44.54 ], [ 116.80 ± 32.87 ], P 〈 0.01 ; [ 60.25 ± 8.73 ] vs [ 132.50 ± 12.15 ], [ 119.20 ± 25.13 ], P 〈 0.01 ）. Conclusion： The expression of antisense CatB RNA can inhibit the growth, adhesion, invasion and migration abilities of MDA-MB-231 cell in vitro.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2010年第2期161-167,共7页
Chinese Journal of Cancer Biotherapy
基金
福建省自然科学基金资助项目(No.X0650058)
福建省属高校项目(No.2006F5047)~~
