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RNA介导的锌指蛋白91假基因敲低对胰腺癌细胞生长和侵袭的抑制作用

Inhibitory effect of RNA-mediated knockdown of zinc finger protein 91 pseudogene on pancreatic cancer cell growth and invasion.

作者信息

Huang Weiyi, Li Ning, Hu Jiong, Wang Lei

机构信息

Department of Oncology, Shanghai Jiaotong University Affiliated Shanghai General Hospital, Shanghai 200085, P.R. China.

出版信息

Oncol Lett. 2016 Aug;12(2):1343-1348. doi: 10.3892/ol.2016.4794. Epub 2016 Jun 28.

Abstract

Worldwide, human pancreatic cancer is a rare malignancy with a poor prognosis. Long non-coding RNAs (lncRNAs) are known to have a crucial role in cancer occurrence and progression; however, the role of pseudogene-expressed lncRNAs, a major type of lncRNA, have not been thoroughly analyzed in cancer. Therefore, the present study focused on zinc finger protein 91 pseudogene (-P). -P expression was initially detected in two pancreatic cancer cell lines by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and the highest expression of -P was found in the BXPC-3-H cell line. Subsequently, BXPC-3-H cells were transfected with -P short hairpin RNA (shRNA) using a plasmid vector and termed sh-P. Cells transfected with negative control plasmid vector were termed shCon. MTT and Transwell assays were performed to analyze the proliferation and migration of BXPC-3-H cells, respectively, and western blotting was used to detect epithelial-mesenchymal transition markers, including vimentin and β-catenin. The present study showed that depletion of -P markedly decreased pancreatic cancer cell proliferation and inhibited cell migration capacity. In addition, the expression of β-catenin increased while vimentin expression decreased. The current findings suggest that high expression of -P promotes the migration of BXPC-3-H cells and may be a novel marker for early diagnosis for pancreatic cancer.

摘要

在全球范围内,人类胰腺癌是一种预后较差的罕见恶性肿瘤。已知长链非编码RNA(lncRNA)在癌症的发生和发展中起关键作用;然而,作为lncRNA主要类型之一的假基因表达lncRNA在癌症中的作用尚未得到充分分析。因此,本研究聚焦于锌指蛋白91假基因(-P)。通过逆转录定量聚合酶链反应(RT-qPCR)最初在两种胰腺癌细胞系中检测到-P的表达,并且在BXPC-3-H细胞系中发现-P的表达最高。随后,使用质粒载体将-P短发夹RNA(shRNA)转染至BXPC-3-H细胞,并命名为sh-P。用阴性对照质粒载体转染的细胞命名为shCon。分别进行MTT和Transwell实验以分析BXPC-3-H细胞的增殖和迁移,并使用蛋白质印迹法检测上皮-间质转化标志物,包括波形蛋白和β-连环蛋白。本研究表明,-P的缺失显著降低胰腺癌细胞的增殖并抑制细胞迁移能力。此外,β-连环蛋白的表达增加而波形蛋白的表达降低。目前的研究结果表明,-P的高表达促进BXPC-3-H细胞的迁移,可能是胰腺癌早期诊断的一个新标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8dca/4950789/6310686537bd/ol-12-02-1343-g00.jpg

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