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长链非编码RNA SPRY4-IT1通过抑制非小细胞肺癌中的上皮-间质转化下调MPZL-1来逆转顺铂耐药性。

Long Non-Coding RNA SPRY4-IT1 Reverses Cisplatin Resistance by Downregulating MPZL-1 via Suppressing EMT in NSCLC.

作者信息

Ye Yunyao, Gu Jingyao, Liu Pei, Wang He, Jiang Lihua, Lei Tianyao, Yu Shanxun, Han Gaohua, Wang Zhaoxia

机构信息

Department of Oncology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, People's Republic of China.

Department of Oncology, Taizhou People's Hospital, Taizhou, Jiangsu, People's Republic of China.

出版信息

Onco Targets Ther. 2020 Apr 2;13:2783-2793. doi: 10.2147/OTT.S232769. eCollection 2020.

DOI:10.2147/OTT.S232769
PMID:32308413
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7135170/
Abstract

PURPOSE

Long non-coding RNA (lncRNA) SPRY4 intronic transcript 1 (SPRY4-IT1) is reported to play important roles in the occurrence and development of many tumors. However, the possible role of SPRY4-IT1 in cisplatin (DDP) resistance of non-small-cell lung cancer (NSCLC) remains unclear. The aim of this study is to investigate the functions and molecular mechanisms underlying SPRY4-IT1 of cisplatin resistance in NSCLC.

METHODS

Expression of SPRY4-IT1 was analyzed in A549 and cisplatin-resistant A549/DDP cell lines by quantitative real-time polymerase chain reaction (RT-qPCR). Overexpression techniques were applied to investigate the biological functions of SPRY4-IT1 in cisplatin-resistant A549/DDP cells. The effects of SPRY4-IT1 on proliferation and apoptosis were evaluated using cell counting kit-8 (CCK8) assays, colony formation assay and flow-cytometric analysis. The expressions of epithelial-mesenchymal transition (EMT)-associated proteins, including E-cadherin and Vimentin, were detected by Western blot. Microarray analysis was performed to identify the putative targets of SPRY4-IT1, which were further verified by Western blotting and RT-qPCR. A549/DDP cells transfected with pCDNA-SPRY4-IT1 were injected into nude mice in order to verify the effect of SPRY4-IT1 on cisplatin resistance in vivo.

RESULTS

The present study demonstrated that SPRY4-IT1 expression was decreased in A549/DDP cells compared with parental A549 cells. Upregulation of SPRY4-IT1 suppressed cell proliferation and caused apoptosis of A549/DDP cells both in vitro and in vivo. MPZL-1 was negatively regulated by SPRY4-IT1. Furthermore, upregulation of SPRY4-IT1 and downregulation of MPZL-1 could suppress epithelial-mesenchymal transition (EMT), which was characterized by increased E-cadherin expression and decreased Vimentin expression.

CONCLUSION

Upregulation of SPRY4-IT1 reversed the cisplatin-resistant phenotype of NSCLC partially by downregulating MPZL-1 via inhibiting EMT process.

摘要

目的

据报道，长链非编码RNA（lncRNA）SPRY4内含子转录本1（SPRY4-IT1）在多种肿瘤的发生和发展中发挥重要作用。然而，SPRY4-IT1在非小细胞肺癌（NSCLC）顺铂（DDP）耐药中的可能作用仍不清楚。本研究旨在探讨SPRY4-IT1在NSCLC顺铂耐药中的功能及分子机制。

方法

采用定量实时聚合酶链反应（RT-qPCR）分析A549细胞和顺铂耐药的A549/DDP细胞系中SPRY4-IT1的表达。应用过表达技术研究SPRY4-IT1在顺铂耐药的A549/DDP细胞中的生物学功能。使用细胞计数试剂盒-8（CCK8）检测、集落形成试验和流式细胞术分析评估SPRY4-IT1对细胞增殖和凋亡的影响。通过蛋白质印迹法检测上皮-间质转化（EMT）相关蛋白E-钙黏蛋白和波形蛋白的表达。进行基因芯片分析以鉴定SPRY4-IT1的潜在靶点，并通过蛋白质印迹法和RT-qPCR进一步验证。将转染了pCDNA-SPRY4-IT1的A549/DDP细胞注射到裸鼠体内，以验证SPRY4-IT1在体内对顺铂耐药的影响。

结果

本研究表明，与亲本A549细胞相比，A549/DDP细胞中SPRY4-IT1的表达降低。SPRY4-IT1的上调在体外和体内均抑制了A549/DDP细胞的增殖并导致其凋亡。MPZL-1受SPRY4-IT1负调控。此外，SPRY4-IT1的上调和MPZL-1的下调可抑制上皮-间质转化（EMT），其特征为E-钙黏蛋白表达增加和波形蛋白表达降低。

结论

SPRY4-IT1的上调通过抑制EMT过程下调MPZL-1，部分逆转了NSCLC的顺铂耐药表型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a41e/7135170/a244576b8817/OTT-13-2783-g0006.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a41e/7135170/64a6d2b148e2/OTT-13-2783-g0002.jpg
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