长链非编码RNA-PICSAR通过miR-485-5p/REV3L轴在皮肤鳞状细胞癌中导致顺铂耐药。
Lnc-PICSAR contributes to cisplatin resistance by miR-485-5p/REV3L axis in cutaneous squamous cell carcinoma.
作者信息
Wang Dan, Zhou Xiaoqiang, Yin Jing, Zhou Yang
机构信息
Plastic Surgery Center, Affiliated Hospital of Jiangnan University, Wuxi, Jiangsu, 214000, China.
Department of Dermatology, Medical College of Shaoguan University, Shaoguan, 512026, China.
出版信息
Open Life Sci. 2020 Jul 10;15(1):488-500. doi: 10.1515/biol-2020-0049. eCollection 2020.
BACKGROUND
Dysregulation of long noncoding RNAs (lncRNAs) is associated with drug resistance in multiple cancers. We explored the roles of lncRNA p38 inhibited cutaneous squamous cell carcinoma-associated lincRNA (PICSAR) in cisplatin (DDP) resistance of cutaneous squamous cell carcinoma (CSCC).
METHODS
Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to measure the expression of lnc-PICSAR, miR-485-5p and reversionless 3-like (REV3L) mRNA. The cell counting kit-8 (CCK-8) assay was conducted to evaluate DDP resistance and cell viability. The transwell assay was performed to determine cell migration and invasion. Western blot assay and immunohistochemistry (IHC) staining assay were carried out to measure protein levels. The dual-luciferase reporter assay was used to investigate the association between miR-485-5p and lnc-PICSAR or REV3L. Murine xenograft model was constructed to explore the function of lnc-PICSAR . The morphology of exosomes was analyzed by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA).
RESULTS
Lnc-PICSAR was elevated in DDP-resistant CSCC cells. Lnc-PICSAR silencing suppressed cell viability, DDP resistance, migration and invasion in DDP-resistant CSCC cells. MiR-485-5p acted as a target of lnc-PICSAR, and miR-485-5p inhibition reversed the impacts of lnc-PICSAR silencing on DDP resistance and cell progression in DDP-resistant CSCC cells. Lnc-PICSAR promoted REV3L expression via sponging miR-485-5p. Moreover, REV3L overexpression overturned the effects of lnc-PICSAR on cell progression and DDP resistance. Lnc-PICSAR knockdown suppressed DDP resistance . In addition, lnc-PICSAR was increased in the exosomes derived from CSCC patients' serum and CSCC cells.
CONCLUSION
Lnc-PICSAR enhanced DDP resistance via miR-485-5p/REV3L axis in DDP-resistant CSCC cells. Besides, exosome-mediated lnc-PICSAR might be involved in the regulation of drug resistance in CSCC.
背景
长链非编码RNA（lncRNA）失调与多种癌症的耐药性相关。我们探讨了长链非编码RNA p38抑制皮肤鳞状细胞癌相关长链间充质RNA（PICSAR）在皮肤鳞状细胞癌（CSCC）顺铂（DDP）耐药中的作用。
方法
采用定量实时聚合酶链反应（qRT-PCR）检测lnc-PICSAR、miR-485-5p和无回复3样（REV3L）mRNA的表达。使用细胞计数试剂盒-8（CCK-8）试验评估DDP耐药性和细胞活力。进行Transwell试验以确定细胞迁移和侵袭能力。采用蛋白质免疫印迹法和免疫组织化学（IHC）染色法检测蛋白质水平。利用双荧光素酶报告基因试验研究miR-485-5p与lnc-PICSAR或REV3L之间的关系。构建小鼠异种移植模型以探究lnc-PICSAR的功能。通过透射电子显微镜（TEM）和纳米颗粒跟踪分析（NTA）分析外泌体的形态。
结果
lnc-PICSAR在DDP耐药的CSCC细胞中表达升高。lnc-PICSAR沉默可抑制DDP耐药CSCC细胞的活力、DDP耐药性、迁移和侵袭能力。miR-485-5p是lnc-PICSAR的靶标，抑制miR-485-5p可逆转lnc-PICSAR沉默对DDP耐药CSCC细胞的DDP耐药性和细胞进展的影响。lnc-PICSAR通过吸附miR-485-5p促进REV3L表达。此外，REV3L过表达可逆转lnc-PICSAR对细胞进展和DDP耐药性的影响。lnc-PICSAR敲低可抑制DDP耐药性。此外，在CSCC患者血清和CSCC细胞来源的外泌体中lnc-PICSAR增加。
结论
在DDP耐药的CSCC细胞中，lnc-PICSAR通过miR-485-5p/REV3L轴增强DDP耐药性。此外,外泌体介导的lnc-PICSAR可能参与CSCC耐药性的调控。
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