lncRNA NEAT1 promoting the proliferation and apoptosis escape of laryngeal papilloma cells by activating BZW2 through the inhibition of miR-let-7a

ZHANG Bo; WANG Song; Ayiheng· Qukuerhan

doi:10.11659/jjssx.10E022108

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lncRNA NEAT1 promoting the proliferation and apoptosis escape of laryngeal papilloma cells by activating BZW2 through the inhibition of miR-let-7a

Basic Research | 更新时间：2023-05-26

- lncRNA NEAT1 promoting the proliferation and apoptosis escape of laryngeal papilloma cells by activating BZW2 through the inhibition of miR-let-7a
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 32, Issue 5, Pages: 382-388(2023)
- 作者机构：
  
  新疆医科大学第一附属医院耳鼻喉科，新疆乌鲁木齐 830054
- 作者简介：
- 基金信息：
- DOI：10.11659/jjssx.10E022108
  CLC： R739.65
- Published：25 May 2023，
  
  Received：21 October 2022，
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张波,王松,阿依恒·曲库尔汗 .lncRNA NEAT1通过抑制miR-let-7a激活BZW2促进喉乳头状瘤细胞的增殖和凋亡逃逸[J].局解手术学杂志,2023,32(05):382-388.

ZHANG Bo,WANG Song,Ayiheng·Qukuerhan .lncRNA NEAT1 promoting the proliferation and apoptosis escape of laryngeal papilloma cells by activating BZW2 through the inhibition of miR-let-7a[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(05):382-388.
张波,王松,阿依恒·曲库尔汗 .lncRNA NEAT1通过抑制miR-let-7a激活BZW2促进喉乳头状瘤细胞的增殖和凋亡逃逸[J].局解手术学杂志,2023,32(05):382-388. DOI： 10.11659/jjssx.10E022108.

ZHANG Bo,WANG Song,Ayiheng·Qukuerhan .lncRNA NEAT1 promoting the proliferation and apoptosis escape of laryngeal papilloma cells by activating BZW2 through the inhibition of miR-let-7a[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(05):382-388. DOI： 10.11659/jjssx.10E022108.

摘要

目的

探讨长链非编码RNA核富集转录本1（lncRNA NEAT1）对喉乳头状瘤（LP）细胞的增殖和凋亡逃逸的调控作用和机制。

方法

培养人喉乳头状瘤细胞Hs840.T，将正常人口腔上皮细胞（HOECs）作为对照。将Hs840.T分为lncRNA NEAT1过表达载体（pcDNA-NEAT1）组、pcDNA-NEAT1阴性对照（pcDNA-NC）组、沉默lncRNA NEAT1的小干扰RNA载体（siNEAT1）组、siNEAT1的阴性对照（siNC）组、miR-let-7a的模拟物（mimic）组、mimic阴性对照（mimic-NC）组、miR-let-7a抑制剂（inhibitor）组、inhibitor阴性对照（inhibitor-NC）组、沉默碱性亮氨酸拉链和W2结构域2（BZW2）的小干扰RNA载体（siBZW2）组、siBZW2的阴性对照（siBZW2-NC）组、pcDNA-NEAT1联合siBZW2给药（pcDNA-NEAT1+siBZW2）组、pcDNA-NEAT1联合siBZW2-NC给药（pcDNA-NEAT1+siBZW2-NC）组。qRT-PCR检测各组中lncRNA NEAT1及其预测靶分子miR-let-7a的表达。Western blot和qRT-PCR检测BZW2的表达。双荧光素酶报告基因实验检测Hs840.T中lncRNA NEAT1和miR-let-7a以及miR-let-7a和BZW2的靶向关系。MTT实验测定Hs840.T的增殖能力。Annexin V-FITC/PI双染法检测Hs840.T的凋亡逃逸能力。

结果

Hs840.T中的lncRNA NEAT1和BZW2表达高于HOECs（

0.05），而miR-let-7a表达低于HOECs（

0.05）。双荧光素酶报告基因实验结果表明，Hs840.T中lncRNA NEAT1吸附抑制miR-let-7a并上调BZW2的表达（

0.05），且BZW2是miR-let-7a的靶基因。与pcDNA-NC组相比，pcDNA-NEAT1组Hs840.T的增殖率增加（

0.05），早期、晚期凋亡率均降低（

0.05）；与siNC组相比，siNEAT1组Hs840.T的增殖率降低（

0.05），早期、晚期凋亡率均增加（

0.05）。另外，与pcDNA-NEAT1+siBZW2-NC组相比，pcDNA-NEAT1+siBZW2组Hs840.T的增殖率降低（

0.05），早期、晚期凋亡率均增高（

0.05）。

结论

lncRNA NEAT1通过抑制miR-let-7a激活BZW2促进LP细胞的增殖和凋亡逃逸。

Abstract

Objective

To investigate the regulatory effect of long non-coding RNA nuclear enriched abundant transcript 1 (lncRNA NEAT1) on the proliferation and apoptosis escape of laryngeal papilloma (LP) cells and its mechanism.

Methods

Human laryngeal papilloma cells Hs840.T were cultured

and the normal human oral epithelial cells (HOECs) were used as the control.Then

the Hs840.T were divided into the lncRNA NEAT1 over-expression vector (pcDNA-NEAT1) group

pcDNA-NEAT1 negative control (pcDNA-NC) group

lncRNA NEAT1 silencing small interfering RNA vector (siNEAT1) group

siNEAT1 negative control (siNC) group

mimic of miR-let-7a (mimic) group

mimic negative control (mimic-NC) group

inhibitor of miR-let-7a (inhibitor) group

inhibitor negative control (inhibitor-NC) group

basic leucine zipper and W2 domains 2 (BZW2) silencing small interfering RNA vector (siBZW2) group

siBZW2 negative control (siBZW2-NC) group

combined administration of pcDNA-NEAT1 and siBZW2 (pcDNA-NEAT1+siBZW2) group

and combined administration of pcDNA-NEAT1 and siBZW2-NC (pcDNA-NEAT1+siBZW2-NC) group.qRT-PCR was used to detect the expression of lncRNA NEAT1 and its predicted target molecule miR-let-7a in each group.Western blot and qRT-PCR were used to detect the expression of BZW2.Dual-luciferase reporter gene assay was used to investigate the targeting relationship between lncRNA NEAT1 and miR-let-7a and the targeting relationship between miR-let-7a and BZW2 in Hs840.T.MTT assay was used to determine the proliferation ability of Hs840.T.Annexin V-FITC/PI double staining assay was used to determine the apoptosis escape ability of Hs840.T.

Results

The expression of lncRNA NEAT1 and BZW2 in Hs840.T were higher than those in HOECs (

0.05)

while the expression of miR-let-7a in Hs840.T was lower than that in HOECs (

0.05).The results of dual-luciferase reporter gene assay showed that lncRNA NEAT1 sponged and inhibited miR-let-7a

thereby up regulating the expression of BZW2 in Hs840.T (

0.05)

and BZW2 was a target gene of miR-let-7a.Compared with the pcDNA-NC group

the pcDNA-NEAT1 group showed a higher proliferation rate of Hs840.T(

0.05)

and lower early and later apoptosis rates (

0.05);compared with the siNC group

the siNEAT1 group showed a lower proliferation rate of Hs840.T (

0.05)

and higher early and later apoptosis rates (

0.05).In addition

compared with the pcDNA-NEAT1+siBZW2-NC group

the pcDNA-NEAT1+siBZW2 group showed a lower proliferation rate of Hs840.T (

0.05)

and higher early and later apoptosis rates (

0.05).

Conclusion

lncRNA NEAT1 promotes the proliferation and apoptosis escape of LP cells by activating BZW2 through the inhibition of miR-let-7a.

关键词

喉乳头状瘤长链非编码RNA核富集转录本1miR-let-7a碱性亮氨酸拉链和W2结构域2细胞凋亡细胞增殖

Keywords

laryngeal papillomalong non-coding RNA nuclear enriched abundant transcript 1miR-let-7abasic leucine zipper and W2 domains 2cell apoptosiscell proliferation

references

Kurita T，Chitose S，Sato K，et al.Pathological mechanisms of laryngeal papillomatosis based on laryngeal epithelial characteristics［J］.Laryngoscope Investig Otolaryngol，2019，4（1）：89-94.doi：10.1002/lio2.242http://dx.doi.org/10.1002/lio2.242.

Matsuzaki H，Makiyama K，Hirai R，et al.Multi‐year effect of human papillomavirus vaccination on recurrent respiratory papillomatosis［J］.Laryngoscope，2020，130（2）：442-447.doi：10.1002/lary.27993http://dx.doi.org/10.1002/lary.27993.

Ouda AM，Elsabagh AA，Elmakaty IM，et al.HPV and recurrent respiratory papillomatosis：a brief review［J］.Life，2021，11（11）：1279-1285.doi：10.3390/life11111279http://dx.doi.org/10.3390/life11111279.

Fernandes JCR，Acuña SM，Aoki JI，et al.Long non-coding RNAs in the regulation of gene expression：physiology and disease［J］.Noncoding RNA，2019，5（1）：17-23.doi：10.3390/ncrna5010017http://dx.doi.org/10.3390/ncrna5010017.

Wang L，Ren C，Xu Y，et al.The LINC00922 aggravates ovarian cancer progression via sponging miR-361-3p［J］.J Ovarian Res，2021，14：77.doi：10.1186/s13048-021-00828-7http://dx.doi.org/10.1186/s13048-021-00828-7.

Dong H，Hu J，Zou K，et al.Correction to：activation of LncRNA TINCR by H3K27 acetylation promotes Trastuzumab resistance and epithelial-mesenchymal transition by targeting MicroRNA-125b in breast cancer［J］.Mol Cancer，2021，20（1）：87.doi：10.1186/s12943-021-01385-9http://dx.doi.org/10.1186/s12943-021-01385-9.

Su Z，Chen M，Ding R，et al.Long noncoding RNA HCG11 suppresses the malignant phenotype of nonsmall cell lung cancer cells by targeting a miR875/SATB2 axis［J］.Mol Med Rep，2021，24（2）：552.doi：10.3892/mmr.2021.12191http://dx.doi.org/10.3892/mmr.2021.12191.

Han L，Zheng C，Wu S.Long noncoding RNA NEAT1 promotes the malignancy of laryngeal squamous cell carcinoma by regulating the microRNA2045p/SEMA4B axis［J］.Oncol Lett，2021，22（5）：1-16.doi：10.3892/ol.2021.13063http://dx.doi.org/10.3892/ol.2021.13063.

Shishodia G，Verma G，Srivastava Y，et al.Deregulation of microRNAs Let-7a and miR-21 mediate aberrant STAT3 signaling during human papillomavirus-induced cervical carcinogenesis：role of E6 oncoprotein［J］.BMC Cancer，2014，23（14）：996.doi：10.1186/1471-2407-14-996http://dx.doi.org/10.1186/1471-2407-14-996.

Liu L，Zhao J，Peng Y，et al.MiR-let-7a-5p inhibits invasion and migration of hepatoma cells by regulating BZW2 expression ［J］.Onco Targets Ther，2020，13（1）：12269-12279.doi：10.2147/OTT.S278954http://dx.doi.org/10.2147/OTT.S278954.

Geng F，Jia WC，Li T，et al.Knockdown of lncRNA NEAT1 suppresses proliferation and migration，and induces apoptosis of cervical cancer cells by regulating the miR377/FGFR1 axis［J］.Mol Med Rep，2022，25（1）：10.doi：10.3892/mmr.2021.12526http://dx.doi.org/10.3892/mmr.2021.12526.

Zhao D，Hou Y.Long non-coding RNA nuclear-enriched abundant transcript 1 （LncRNA NEAT1） upregulates Cyclin T2 （CCNT2） in laryngeal papilloma through sponging miR-577/miR-1224-5p and blocking cell apoptosis［J］.Bioengineered，2022，13（1）：1828-1837.doi：10.1080/21655979.2021.2017653http://dx.doi.org/10.1080/21655979.2021.2017653.

Liu J，Yang T，Zhang Y，et al.Promotion of BZW2 by LINC00174 through miR-4500 inhibition enhances proliferation and apoptosis evasion in laryngeal papilloma［J］.Cancer Cell Int，2020，20（1）：471.doi：10.1186/s12935-020-01559-3http://dx.doi.org/10.1186/s12935-020-01559-3.

Fan JT，Zhou ZY，Luo YL，et al.Exosomal lncRNA NEAT1 from cancer-associated fibroblasts facilitates endometrial cancer progression via miR-26a/b-5p-mediated STAT3/YKL-40 signaling pathway［J］.Neoplasia，2021，23（7）：692-703.doi：10.1016/j.neo.2021.05.004http://dx.doi.org/10.1016/j.neo.2021.05.004.

Shen X，Ye Z，Wu W，et al.lncRNA NEAT1 facilitates the progression of colorectal cancer via the KDM5A/Cul4A and Wnt signaling pathway［J］.Int J Oncol，2021，59（1）：51.doi：10.3892/ijo.2021.5231http://dx.doi.org/10.3892/ijo.2021.5231.

Qiao Y，Jin T，Guan S，et al.Long non-coding RNA Lnc-408 promotes invasion and metastasis of breast cancer cell by regulating LIMK1［J］.Oncogene，2021，40（24）：4198-4213.doi：10.1038/s41388-021-01845-yhttp://dx.doi.org/10.1038/s41388-021-01845-y.

Yu T，Yu J，Lu L，et al.MT1JP-mediated miR-24-3p/BCL2L2 axis promotes Lenvatinib resistance in hepatocellular carcinoma cells by inhibiting apoptosis［J］.Cell Oncol （Dordr），2021，44（4）：821-834.doi：10.1007/s13402-021-00605-0http://dx.doi.org/10.1007/s13402-021-00605-0.

Duan YR，Chen BP，Chen F，et al.LncRNA lnc-ISG20 promotes renal fibrosis in diabetic nephropathy by inducing AKT phosphorylation through miR-486-5p/NFAT5［J］.J Cell Mol Med，2021，25（11）：4922-4937.doi：10.1111/jcmm.1628http://dx.doi.org/10.1111/jcmm.1628.

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