摘要
目的:探索富脯氨酸蛋白11(Proline-rich protein 11,PRR11)过表达诱发染色体凝聚异常的分子机理。方法:综合利用瞬时过表达、si RNA敲减和免疫荧光染色方法在人肺癌细胞系A549中过表达或沉默PRR11后,检测波形蛋白(Vimentin)的表达与亚细胞分布及染色质凝聚异常情况。结果:肺癌细胞中内源性PRR11在胞浆中呈现弥散型分布,而Flag-PRR11过表达后在细胞中的分布呈现弥散状和网络状两种不同形态。进一步检测发现内源性或外源过表达PRR11与Ⅲ型中间丝蛋白Vimentin共定位,并且导致Vimentin绕核分布紊乱及染色质凝聚异常。结论:在细胞周期过程中PRR11与Vimentin共定位并调节其组装过程。推测PRR11异常表达可能通过与Vimentin共定位导致核骨架核纤层不稳定并诱发染色质异常凝聚,从而促进肿瘤的发生发展。
Objective ：To explore the molecular mechanism of over-expression of Proline-rich protein 11 （PRR11） gene inducing ab- normal chromatin condensation. Methods：The PRR11 gene was down-regulated and over-expressed using siRNA transient over-ex- pression and immunofluorescent staining in lung cancer cell A549. The expressions of the Vimentin at mRNA and protein levels were detected by RT-PCR and Western blot after the PRR11 treatment,respectively. Subcellular distribution of Vimentin and disruption of chromatin condensation was detected by immunofluorescence. Results：Endogenous PRR11 proteins were dispersively distributed in cytoplasm, and over-expressed Flag-PRR11 proteins were dispersively and reticularly arranged in cytoplasm. The results showed that both endogenous and exogenous over-expressed PRR11 proteins were co-localized with Vimentin. Disorder arrangement of Vimentin in the perinuclear area and disruption of chromatin condensation were observed after over expressing PRR11 proteins. Conclusion：The PRR11 proteins are co-localized with Vimentin, and the distribution of Vimentin proteins is re-arranged after forced expression of PRR11 proteins in lung cancer cell A549. We speculate that abnormal expression of PRR11 proteins could induce the instability of Lamins and abnormal chromatin condensation through co-localization with Vimentin and thereby promotes the development of tumor.
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2016年第7期733-737,共5页
Journal of Chongqing Medical University
基金
国家自然科学基金资助项目(编号:81501979)
重庆市自然科学基金计划资助项目(编号:cstc2013jcyjA10043)
