【摘要】 目的探讨CD44单链抗体sc Fv CD44、肿瘤坏死因子相关凋亡诱导配体(TRAIL)双基因共表达真核载体(p IRES-sc Fv CD44-TRAIL)对人乳腺癌MDA-MB-231细胞株的抑制作用。方法 1细胞实验:采用RT-PCR二步法构建p IRES-sc Fv CD44-TRAIL并转染MDA-MB-231细胞(转染组);设转染p IRES空质粒的空白组及不作任何处理的阴性组。转染24 h后采用流式细胞仪检测各组细胞凋亡情况,计算细胞凋亡率;行划痕试验比较各组细胞迁移情况,计算划痕愈合率。2移植瘤实验:建立乳腺癌荷瘤裸鼠模型8只并随机分为实验组及对照组。实验组皮下注射p IRES-sc Fv CD44-TRAIL 100μL/只,隔日1次,共5次;对照组注射等量生理盐水。观察两组移植瘤生长情况,计算抑瘤率;采用免疫组化法检测两组瘤组织中Ki67的表达水平。结果 1细胞实验:转染组细胞凋亡率明显高于、划痕愈合率明显低于空白组和阴性组。2移植瘤实验:观察组抑瘤率为79.8%;观察组Ki67的阳性细胞率明显低于对照组,P<0.05。结论 p IRES-sc Fv CD44-TRAIL可抑制人乳腺癌细胞及其移植肿瘤的生长及转移。更多还原

【Abstract】 Objective To investigate the inhibitory effects of CD44single-chain antibody sc Fv CD44 and tumor necrosis factor-related apoptosis inducing ligand eukaryotic expression vector( p IRES-sc Fv CD44-TRAIL) on breast cancer cells MDA-MB-231. Methods 1 Cell experiments: The plasmid DNA of p IRES-sc Fv CD44-TRAIL,whose genes were amplified by reverse transcriptase-polymerase chain reaction( RT-PCR),was transfected into breast cancer cells MDA-MB-231( the transfected group). The blank group was transfected with the blank p IRES plasmid and the negative group was not treated. After 24 hours,the apoptosis rate was calculated by flow cytometry. The scratch test was performed to observe the cell motility assay as well as to calculate the scratch healing rate. 2 Graft model experiments: Eight nude mice of breast cancer graft model were randomly divided into the experiment group and the control group. Mice in the experiment group received subcutaneous injection of p IRES-sc Fv CD44-TRAIL 100μL per rat,once two days,and for five times totally. Mice in the control group were treated by the same amount of normal saline. Tumor growth was observed,and the tumor inhibitory rate was calculated. The expression of Ki67 was detected by immunohistochemistry. Results 1 Cell experiments: The transfected group had higher apoptosis rate and lower wound-healing rate as compared with the blank group and the negative group. 2 Graft model experiments: The tumor inhibitory rate of the observation group was 79. 8%. The Ki67-positive rate of the observation group was significantly lower than that of the control group( P < 0. 05). Conclusion p IRES-sc Fv CD44-TRAIL can inhibit the growth and metastasis of breast cancer cells and its transplanted tumor.更多还原