BLACAT3募集hnRNPK调控pre-mRNA可变剪接促进膀胱癌抗失巢凋亡和转移

Anoikis resistance plays akey role in the survival and metastasis of bladder cancer. We have screened and identified a long noncoding RNA BLACAT3 (bladder cancer associated transcript 3) from our previous microarray results which enhance the anoikis resistance and metastatic capability of bladder cancer cell. It suggests that BLACAT3 is able to assist anti-apoptosis and promote metastasis of bladder cancer cells via binding with hnRNPK (heterogeneous nuclear ribonucleoprotein k) to mediate spliceosome transformation of BCL-X and P120CTN. Hence, our study will 1) verify the capability of anoikis resistance and pro-metastasis of BLACAT3 in vitro and in vivo; 2) explore the mechanisms of BLACAT3 that wheather BLACAT3 regulate alternative splicing of BCL-X and P120CTN through recruiting hnRNPK by performing RNA and protein immunoprecipitation. In a word, based on the hypothesis of “BLACAT3 promotes anoikis resistance and metastasis of bladder cancer by recruiting hnRNPK”, our study focuses mainly on the experimental evidences for the anti-apoptotic and prognostic role of BLACAT3 in bladder cancer, which may suggest novel strategies for the metastatic prognosis and clinical treatment on BLACAT3 in bladder cancer.

抗失巢凋亡对膀胱癌细胞播散及存活至关重要。我们前期基因芯片数据筛选鉴定出增强膀胱癌细胞抗失巢凋亡能力的长链非编码RNA-BLACAT3，其可能通过结合异质性核糖蛋白K（hnRNPK）介导BCL-X、P120CTN基因的剪接体转换，赋予膀胱癌细胞抗凋亡表型和转移特征。本项目拟在体内外模型证实BLACAT3过表达促进膀胱癌细胞抗失巢凋亡和转移，应用RNA免疫沉淀、蛋白质免疫共沉淀等实验阐明BLACAT3募集并引导hnRNPK调控BCL-X、P120CTN等下游基因mRNA前体选择性剪接的分子机制。本项目围绕“BLACAT3直接结合hnRNPK调控pre-mRNA可变剪接促进膀胱癌抗失巢凋亡和转移”这一科学假说，通过干预BLACAT3及其靶基因获得它们调控失巢凋亡和转移的实验依据，明确BLACAT3与患者预后的关系，为BLACAT3作为预测膀胱癌转移的分子标记和寻找治疗新靶点提供原创性科学依据。

抗失巢凋亡对膀胱癌细胞播散及存活至关重要。我们前期基因芯片数据筛选鉴定出增强膀胱癌细胞抗失巢凋亡能力的长链非编码RNA-BLACAT3，其可能通过结合异质性核糖蛋白K（hnRNPK）介导BCL-X、P120CTN基因的剪接体转换，赋予膀胱癌细胞抗凋亡表型和转移特征。本项目通过体内外模型证实BLACAT3过表达促进膀胱癌细胞抗失巢凋亡和转移，初步阐明BLACAT3募集并引导hnRNPK调控BCL-X、P120CTN等下游基因mRNA前体选择性剪接的分子机制。初步证实了BLACAT3与患者预后的关系，为BLACAT3作为预测膀胱癌转移的分子标记和寻找治疗新靶点提供原创性科学依据。