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作 者:陶光晶 严奉奇 马柳疆 李霞[3] 袁建林[2] 武国军[2]
机构地区:[1]安康市中心医院泌尿外科,陕西安康725000 [2]第四军医大学西京医院泌尿外科,陕西西安710032 [3]第四军医大学生物化学与分子生物学教研室,陕西西安710032
出 处:《现代肿瘤医学》2016年第20期3180-3184,共5页Journal of Modern Oncology
基 金:国家自然科学基金资助项目(编号:81272812)
摘 要:目的:利用质粒转染技术制备过表达迁移侵袭抑制蛋白(migration and invasion inhibitory protein,MIIP)的肾癌786-0/MIIP细胞系,观察MIIP对细胞增殖、侵袭及迁移能力的影响。方法:构建携带MIIP基因的质粒,通过脂质体质粒转染技术上调肾癌786-0细胞系MIIP基因表达。Western-blot、Real-time PCR检测转染效果;利用MTT实验检测MIIP对786-0细胞增殖能力的影响;Transwell实验、划痕实验检测MIIP对786-0细胞侵袭、迁移能力的影响。结果:Western-blot、Real-time PCR检测发现实验组786-0/MIIP细胞MIIP蛋白及mRNA表达量有效上调。MTT实验显示实验组786-0/MIIP细胞增殖能力明显减弱(P<0.01)。Transwell实验显示实验组786-0/MIIP细胞侵袭能力减弱,穿透小室细胞数明显减少(P<0.01)。细胞划痕实验显示实验组786-0/MIIP细胞迁移能力显著下降(P<0.01)。结论:通过质粒转染技术能够制备过表达MIIP的肾癌细胞系。转染成功的肾癌786-0/MIIP细胞系MIIP表达量增加,有效抑制了肾癌细胞的增殖、侵袭与迁移。Objective :To prepare renal carcinoma cell line 786 -0/MIIP via plasmid transfection technique, then investigate the change of renal carcinoma cellg proliferation, invasion and migration ability. Methods :The renal carci- noma cell strain 786 -0 was transfeeted by MIIP gene plasmid. The transfeetion effects were detected by Real -time PCR and Western- blot technique. The change of eellg proliferation, invasion and metastasia ability was detected by MTT assay,Transwell assay and ground -healing assay respectively. Results: By the Western -bolt and Real -time PCR technique, the expression levels of MIIP protein and mRNA in 786 -0/MIIP cell strain were up - regulated effectively. Meanwhile, the proliferation and invasion ability of 786 - 0/MIIP group decreased significantly( P 〈 0.01 ) , and the number of 786 -0/MIIP cell that had through the membrane was the lowest in the test(P 〈 0.01 ). Similarly, the migration distance of 786 - 0/MIIP cell was the shortest in Wound - healing assay-(P 〈 0.01 ). Conclusion: Through the plasmid transfection technique,the RCC cell line that could over expressed MIIP were prepared successfully and it can suppres the renal cancer eellg proliferation,invasion and metastasis.
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