骨桥蛋白不同剪接变体对肺癌细胞生长、侵袭及转移的影响  

The effects of splicing variants of osteopontin on cell growth,migration and invasion of lung cancer cells

作　　者：宋水川[1] 陈达伟[1] 江晓肖[1] 

机构地区：[1]中国人民解放军117医院消化科,浙江杭州310013

出　　处：《现代肿瘤医学》2014年第3期529-533,共5页Journal of Modern Oncology

摘　　要：目的:研究骨桥蛋白(OPN)不同剪接变体在三株人肺癌细胞系A549、NCI-H209和95-D中的表达情况及其对肺癌细胞增殖、侵袭转移的影响。方法:RT-PCR和Western blot方法检测OPN不同剪接变体在三株肺癌细胞系中的mRNA和蛋白水平。构建不同OPN剪接变体的瞬时转染细胞。软琼脂克隆形成实验、细胞侵袭及划痕实验观察不同OPN剪接变体转染的A549细胞体外增殖、侵袭以及转移能力。结果:在三株肺癌细胞系中,OPN有3种剪接变体形式表达,A549和NCI-H209细胞中OPN-a和OPN-b的mRNA水平高于OPN-c,OPN-c的mRNA水平在95-D细胞中最高。Western blot也进一步发现三种剪接变体在三株肺癌细胞系中表达,三种剪接变体的蛋白表达水平与mRNA水平不一致,主要以OPN-a形式高表达。瞬转三种剪接变体至A549细胞,发现瞬转OPN-a和OPN-b可以更有效的促进A549细胞的增殖,而OPN-b和OPN-c对侵袭能力促进更明显,但是OPN-c更有效促进细胞迁移。结论:不同OPN剪接变体形式在肺癌细胞中的表达水平不同,其对肿瘤的生物学行为影响也不同。鉴定特异的OPN剪接变体可更有效的为评价肿瘤生长及转移提供参考指标。Objective ：To investigate the expression of splicing variants of osteopontin （OPN - a,OPN - b, OPN - c） on lung cancer cell line A549,NCI - 11209 and 95 - D, and the role of these isoforln on tumor cell growth,migra- tion and invasion in vitro. Methods：The relative expression of OPN splicing variants in human lung cancer cell lines was determined by RT - PCR and Western blot. Lentivirus vector carried OPN splicing variants was transfected into A549 cells transietly, cell growth,invasion and migration were determined by softagar colony forlnation assay,Matrigel transwell assay,cell wound healing assay. Results： High mRNA and protein expressions of OPN - a, OPN - b and OPN - e were detected in three lung cancer cell lines. Among them,the levels of OPN - a and OPN - b in A549, NCI - 14209 cells showed higher than the level of OPN - e,but higher expression of OPN - e was detected in 95 - D ceils. in protein levels, OPN - a was mainly expressed in three lung cancer cell lines, and the protein levels of three splice- some were not consistent with mRNA levels. Furthermore,over - expression of OPN - a and OPN - b in A549 cells enhanced cell growth, and OPN - e showed no obvious effect on cell growth. However, over - expression of OPN - b and OPN - e significantly increased cell invasion, compared with OPN - a transfection and mock groups. Furthermore, we found that over - expression of three splicing variants of OPN promoted cell migration, but OPN - e showed more effect,compared with other groups. Conclusion：Splicing variants of OPN were differential expressed in lung cancer cell, and play different roles in tumor cell growth, invasion and migration.

关 键 词：肺癌细胞 骨桥蛋白 剪接变体 侵袭转移 

分 类 号：R734.2[医药卫生—肿瘤]