HnRNPA2B1在乳腺癌细胞MCF7中的作用与机研究.docx

摘要上述研究结果表明，敲除hnRNPA2fBl会抑制乳腺癌细胞的增殖，促进其迁 摘要 上述研究结果表明，敲除hnRNPA2fBl会抑制乳腺癌细胞的增殖，促进其迁 移和侵袭，抑制其凋亡和自噬，而且hnRNPA2／B1与STAT3、NPM有相互作用。 我们的结果提示hnRNPA2／B1可能在乳腺癌中的作用并不简单的是肿瘤抑制或 促进作用。HnRNPA2／B1在肿瘤的进程中可能起着双重的作用，其促进了细胞的 增殖，但抑制了细胞的迁移和侵袭。HnRNPA2／B1的表达水平可能成为乳腺癌进 展及预后的一个潜在标志物。 关键词： CRISPR—Cas9；hnRNPA2／B1；相互作用蛋白网络；细胞迁移；乳腺癌 万方数据 AbnraorionAbstraction Abnraorion Abstraction An emerging body of data shows the overexpression of hnRNPA2／B 1 in many cancers，but its specific molecular mechanism in tumors is still poorly understand．In order to explore the mechanism of hnRNPA2／B 1 and fmd valid therapeutic target in breast cancer．We used CRISPR．Cas9 to construct the stable MCF一7 cells in which hnRNPA2／B 1 were konck-out，and explored the change of proliferation，migration， invasion，apoptosis，autophagy．Besides，affmity purification by flag magnetic beads and identified the separated proteins by HPLC—MS．Bioinformatics analysis of MS results using string database and costructed the interation network of hnRNPB 1，the KEGG pathway of the interation network of hnRNPB 1。 MTT assay showed knock—out of hnRNPA2／B 1 reduce the proliferation of MCF．7 cells，transwell assay results showed knock—out of hnRNPA2／B 1 increased the migration and invasion of MCF-7 cells．besides knock—out of hnRNPA2／B 1 inhibited apoptosis and autophagy．Western blotting results showed that the expression of P-STAT3，AKT,P．AKT,CDK4 decreased and the expression of cell cycle inhibitor P27 increased．Some genes related to EMT,for example，the expression of E．Cadherindecreased，theexpressionofsnail，vimentin，twistincreased．Somegenes related to apoptosis，for instance，the expression of Bax decreased，the expression of cleaved．caspase3 decreased．Some genes related to autophagy,the expression of LC3 II decreased，the expression of P62 increased．QPCR results showed that knock-out of hnRNPA2／B 1 decreased the mRNA level of E．Cadherin，and increased the mRNA level of snail，vimentin，twist，VEGF-A．Alternative splicing results showed the skipping of 1 1 of RO