[1]胡艳正,牛彦杰,石鹏飞.长链非编码RNA LINC00222对肺腺癌细胞增殖、迁移、侵袭和凋亡的影响及作用机制的研究[J].现代检验医学杂志,2021,36(05):16-22.[doi:10.3969/j.issn.1671-7414.2021.05.004]
 HU Yan-zheng,NIU Yan-jie,SHI Peng-fei.Effect of Long Non Coding RNA LINC00222 on Proliferation, Migration,Invasion and Apoptosis of Lung Adenocarcinoma Cells and Its Mechanism[J].Journal of Modern Laboratory Medicine,2021,36(05):16-22.[doi:10.3969/j.issn.1671-7414.2021.05.004]
点击复制

长链非编码RNA LINC00222对肺腺癌细胞增殖、迁移、侵袭和凋亡的影响及作用机制的研究()
分享到:

《现代检验医学杂志》[ISSN:/CN:]

卷:
第36卷
期数:
2021年05期
页码:
16-22
栏目:
论 著
出版日期:
2021-10-14

文章信息/Info

Title:
Effect of Long Non Coding RNA LINC00222 on Proliferation, Migration,Invasion and Apoptosis of Lung Adenocarcinoma Cells and Its Mechanism
文章编号:
1671-7414(2021)05-016-07
作者:
胡艳正牛彦杰石鹏飞
(咸阳市第一人民医院胸外科,陕西咸阳 712000)
Author(s):
HU Yan-zheng NIU Yan-jie SHI Peng-fei
(Department of Thoracic Surgery, Xianyang First People’s Hospital, Shaanxi Xianyang 712000,China)
关键词:
长链非编码RNAs LINC00222肺腺癌生物学行为
分类号:
R734.2;R730.43
DOI:
10.3969/j.issn.1671-7414.2021.05.004
文献标志码:
A
摘要:
目的 检测肺腺癌组织及细胞系中LncRNA LINC00222表达,探究其对肺腺癌细胞增殖、迁移、侵袭和凋亡的影响和相关作用机制。方法 采用实时荧光定量PCR(qRT-PCR)法和Western blot 实验检测肺腺癌组织及细胞中LINC00222表达;构建LINC00222过表达载体,验证其转染效率;采用CCK-8法、Hoechst 33342/PI 染色法、划痕实验及Transwell实验分别检测过表达LINC00222对肺腺癌细胞增殖、凋亡、迁移和侵袭的影响;采用Western blot 法检测肺腺癌细胞中P-GSK-3β,GSK-3β蛋白及β-catenin核蛋白表达;采用荧光霉素基因实验及RIP实验验证探究LINC00222影响肺腺癌生物学行为的相关作用机制。结果 肺腺癌组织中LINC00222 mRNA和蛋白表达明显低于癌旁正常组织,差异有统计学意义(t=7.388,15.100,均P<0.001);肺腺癌细胞系中LINC00222表达明显低于人正常肺胚细胞(F=21.926,P<0.001)。过表达LINC00222后,肺腺癌细胞增殖、迁移、侵袭能力明显抑制,细胞凋亡数目明显增多(P<0.01)。过表达 LINC00222后,GSK-3β磷酸化明显降低, GSK-3β催化活性明显增强,β-catenin 核转位受到抑制(P<0.01)。LINC00222靶向调控结合GSK-3β和GSK-3β蛋白共沉淀中LINC00222表达显著高于IgG蛋白共沉淀(P<0.05)。结论 肺腺癌中LncRNA LINC00222低表达,其过表达可抑制肺腺癌细胞的增殖、迁移及侵袭,促进细胞凋亡,可能与其调控GSK-3β催化活性,抑制β-catenin 核转位有关。
Abstract:
Objective The expression of lncRNA LINC00222 in lung adenocarcinoma tissues and cell lines was detected toexplore the effects of lncRNA LINC00222 on the proliferation,migration,invasion and apoptosis of lung adenocarcinoma cellsand the related mechanisms. Methods The expression of LINC00222 in lung adenocarcinoma tissues and cells was detected byqRT-PCR and Western blot. Overexpression vector LINC00222 was constructed to verify its transfection efficiency. CCK-8 assay,Hoechst 33342/PI staining assay,scratch assay and Transwell assay were used to detect the effects of overexpression ofLINC00222 on proliferation,apoptosis,migration and invasion of lung adenocarcinoma cells.Western blot was used to detect theexpression of P-GSK-3β,GSK-3βprotein and β-catenin nuclear protein in lung adenocarcinoma cells. Luciferin geneexperiment and RIP experiment were used to investigate the mechanism of LINC00222 affecting the biological behavior of lungadenocarcinoma. Results The mRNA and protein expressions of LINC00222 in lung adenocarcinoma tissues were significantlylower than those in adjacent normal tissues, the difference were statistically significant (t=7.388, 15.100,all P<0.001). Theexpression of LINC00222 in lung adenocarcinoma cell lines was significantly lower than that in normal human lung embryo celllines (F=21.926, P<0.001). After the overexpression of LINC00222,the proliferation, migration and invasion of lungadenocarcinoma cells were significantly inhibited,and the number of cell apoptosis was significantly increased (P<0.01). Afteroverexpression of LINC00222,GSK-3β phosphorylation was significantly decreased,GSK-3β catalytic activity wassignificantly enhanced,and β-catenin nuclear translocation was inhibited (P<0.01). LINC00222 targeted the binding of GSK-3β,and the expression of LINC00222 in GSK-3β protein co-precipitation was significantly higher than that in IgG protein coprecipitation(P< 0.05). Conclusion LncRNA LINC00222 was low expressed in lung adenocarcinoma,and its overexpressioninhibited the proliferation,migration and invasion of lung adenocarcinoma cells,and promoted cell apoptosis,which may be related to its regulation of GSK-3β catalytic activity and inhibition of β-catenin nuclear translocation.

参考文献/References:

[1] BENDELS MICHAEL H K, BR?GGMANN D.SCH?FFEL N, et al. Gendermetrics of cancerresearch:results from a global analysis on lungcancer[J]. Oncotarget, 2017, 8(60): 101911-101921.
[2] SUN Weiliang, YANG Yunben, XU Chunjing, et al.Regulatory mechanisms of long noncoding RNAs ongene expression in cancers[J]. Cancer Genetics, 2017,216-217: 105-110.
[3] ENGREITZ J M, HAINES J E, PEREZ E M, etal. Local regulation of gene expression by lncRNApromoters, transcription and splicing[J]. Nature, 2016,539(7629): 452-455.
[4] DYKES I M, EMANUELI C. Transcriptional and posttranscriptionalgene regulation by long non-codingRNA[J]. Genomics, Proteomics & Bioinformatics,2017, 15(3): 177-186.
[5] CHEN Dong, SUN Qiang, CHENG Xiaofei, et al.Genome-wide analysis of long noncoding RNA(lncRNA) expression in colorectal cancer tissues frompatients with liver metastasis[J]. Cancer Medicine,2016, 5(7): 1629-1639.
[6] XIE Shanshan, JIN Juan, XU Xiao, et al. Emergingroles of non-coding RNAs in gastric cancer:Pathogenesis and clinical implications[J]. World Journalof Gastroenterology, 2016, 22(3): 1213-1223.
[7] DONG Rui, JIA Deshui, XUE Ping, et al. Genomewideanalysis of long noncoding RNA (lncRNA)expression in hepatoblastoma tissues[J]. PLoS One,2014, 9(1): e85599.
[8] SHI Xiuhui, ZHAO Yan, HE Ruizhi, et al. Three-lncRNA signature is a potential prognostic biomarker forpancreatic adenocarcinoma[J]. Oncotarget, 2018, 9(36):24248-24259.
[9] YIN Dandan, LU Xiyi, SU Jun, et al. Long noncodingRNA AFAP1-AS1 predicts a poor prognosis andregulates non-small cell lung cancer cell proliferationby epigenetically repressing p21 expression[J].Molecular Cancer, 2018, 17(1): 92.
[10] ZHANG Yajia, PITCHIAYA S, CIE?LIK M, et al.Analysis of the androgen receptor-regulated lncRNAlandscape identifies a role for ARLNC1 in prostatecancer progression[J]. Nature Genetics, 2018, 50(6):814-824.
[11] ENGREITZ J M, OLLIKAINEN N, GUTTMAN M.Long non-coding RNAs: spatial amplifiers that controlnuclear structure and gene expression[J]. Nature ReviewsMolecular Cell Biology, 2016, 17(12): 756-770.
[12] QUINN J J, CHANG H Y. Unique features of longnon-coding RNA biogenesis and function[J]. NatureReviews Genetics, 2016, 17(1): 47-62.
[13] XI Jie, FENG Jing, ZENG Saitian. Long noncodingRNA lncBRM facilitates the proliferation, migrationand invasion of ovarian cancer cells via upregulation ofSox4[J]. American Journal of Cancer Research, 2017,7(11): 2180-2189.
[14] ZHANG Junyong, ZHANG Di, ZHAO Qi, et al. Adistinctively expressed long noncoding RNA, RP11-466I1.1, May serve as a prognostic biomarker inhepatocellular carcinoma[J]. Cancer Medicine, 2018,7(7): 2960-2968.
[15] LI Shuqin, ZHOU Jun, WANG Zhaoxin, et al. Longnoncoding RNA GAS5 suppresses triple negative breastcancer progression through inhibition of proliferationand invasion by competitively binding miR-196a-5p[J].Biomedicine & Pharmacotherapy, 2018, 104: 451-457.
[16] KOPP F, MENDELL J T. Functional classification andexperimental dissection of long noncoding RNAs[J].Cell, 2018, 172(3): 393-407.
[17] ENGREITZ J M, HAINES J E, PEREZ E M, etal. Local regulation of gene expression by lncRNApromoters, transcription and splicing[J]. Nature, 2016,539(7629): 452-455.
[18] 周征宇, 向颖, 袁帅, 等. 长链非编码RNA LINC00324 对非小细胞肺癌细胞生物学功能的影响[J].第三军医大学学报,2019,41(4):331-338.ZHOU Zhengyu, XIANG Ying, YUAN Shuai, etal. Effect of long non-coding RNA LINC00324on biological functions of non-small cell lungcancer cells [J]. Journal of Third Military MedicalUniversity,2019,41(4):331-338.
[19] CHEN Y G , SATPATHY A T, CHANG H Y. Generegulation in the immune system by long noncodingRNAs[J]. Nature Immunology, 2017, 18(9): 962-972.
[20] ZOU Zigui, MA Tianshi, HE Xuezhi, et al. Longintergenic non-coding RNA 00324 promotes gastriccancer cell proliferation via binding with HuR andstabilizing FAM83B expression[J]. Cell Death &Disease, 2018, 9(7): 717.
[21] PANDEY M K, DEGRADO T R. Glycogen synthasekinase-3 (GSK-3)-Targeted therapy and imaging[J].Theranostics, 2016, 6(4): 571-593.
[22] ZHAN T, RINDTORFF N, BOUTROS M. Wnt signalingin cancer[J]. Oncogene, 2017, 36(11): 1461-1473.
[23] MANCINELLI R, CARPINO G, PETRUNGAROS, et al. Multifaceted roles of GSK-3 in cancer andautophagy-related diseases[J]. Oxidative Medicine andCellular Longevity, 2017,2017: 4629495.
[24] GU Ye, WANG Zhirong, SHI Jiawei, et al. Titaniumparticle-induced osteogenic inhibition and bonedestruction are mediated by the GSK-3β/β-cateninsignal pathway[J]. Cell Death & Disease, 2017, 8(6):e2878.
[25] JAHAN S, SINGH S, SRIVASTAVA A, et al. PKAGSK3βand β-Catenin signaling play a critical rolein Trans-Resveratrol mediated neuronal differentiationin human cord blood stem cells[J]. MolecularNeurobiology, 2018, 55(4): 2828-2839.
[26] JAIN S, GHANGHAS P, RANA C, et al. Role of GSK-3β in regulation of canonical Wnt/β-catenin signalingand PI3-K/Akt oncogenic pathway in colon cancer[J].Cancer Investigation, 2017, 35(7): 473-483.
[27] SUN Chengming, LUAN Shuping, ZHANG Guili,et al. CEBPA-mediated upregulation of the lncRNAPLIN2 promotes the development of chronicmyelogenous leukemia via the GSK3 and Wnt/β-catenin signaling pathways[J]. American Journal ofCancer Research, 2017, 7(5): 1054-1067.

相似文献/References:

[1]刘 欣,张欢欢,关 平,等.肺腺癌细胞中长链非编码 RNA- ARAP1-AS1 表达模式及其生物学特征[J].现代检验医学杂志,2021,36(03):38.[doi:10.3969/j.issn.1671-7414.2021.03.009]
 LIU Xin,ZHANG Huan-huan,GUAN Ping,et al.Expression Pattern and Biological Characteristics of ARAP1-AS1 in LungAdenocarcinoma Cells[J].Journal of Modern Laboratory Medicine,2021,36(05):38.[doi:10.3969/j.issn.1671-7414.2021.03.009]
[2]王利民,刘 锴.TUBA1C促进人肺腺癌细胞系增殖和迁移的机制研究[J].现代检验医学杂志,2021,36(04):74.[doi:10.3969/j.issn.1671-7414.2021.04.016]
 WANG Li-min,LIU Kai.Study on the Mechanism of TUBA1C Promoting Proliferation and Migrationof Lung Adenocarcinoma Cells[J].Journal of Modern Laboratory Medicine,2021,36(05):74.[doi:10.3969/j.issn.1671-7414.2021.04.016]
[3]鲍培龙,徐月亮,王孝彬.miR-153对肺腺癌细胞增殖、侵袭、迁移和凋亡的影响及机制研究[J].现代检验医学杂志,2022,37(01):107.[doi:10.3969/j.issn.1671-7414.2022.01.022]
 BAO Pei-long,XU Yue-liang,WANG Xiao-bin.Effects of miR-153 on Proliferation, Invasion, Metastasis and Apoptosis of Lung Adenocarcinoma Cells and Its Mechanism[J].Journal of Modern Laboratory Medicine,2022,37(05):107.[doi:10.3969/j.issn.1671-7414.2022.01.022]
[4]董胜鉴,黄丽丽,李树锦.肺腺癌患者血清中miR-498,miR-339-5p和miR-210-3p水平表达的临床诊断价值[J].现代检验医学杂志,2022,37(02):58.[doi:10.3969/j.issn.1671-7414.2022.02.012]
 DONG Sheng- jian,HUANG Li- li,LI Shu -jin.Clinical Diagnostic Value Expressed at miR-498, miR-339-5p and miR-210-3p Levels in Serum in Patients with Lung Adenocarcinoma[J].Journal of Modern Laboratory Medicine,2022,37(05):58.[doi:10.3969/j.issn.1671-7414.2022.02.012]
[5]苗 毅a,杨淑梅a,王 晶b,等.肺腺癌组织中MCM4 的表达及其与预后、免疫微环境的相关性分析[J].现代检验医学杂志,2023,38(05):115.[doi:10.3969/j.issn.1671-7414.2023.05.022]
 MIAO Yia,YANG Shumeia,WANG Jingb,et al.Expression of MCM4 in Lung Adenocarcinoma and Its Correlation with Prognosis and Immune Microenvironment[J].Journal of Modern Laboratory Medicine,2023,38(05):115.[doi:10.3969/j.issn.1671-7414.2023.05.022]

备注/Memo

备注/Memo:
基金项目:陕西省社会发展科技攻关项目(2018SF081)。
作者简介:胡艳正(1986-),男,研究生,主治医师,研究方向:肺癌、食管癌、纵隔肿瘤。
通讯作者:石鹏飞(1979-),男,副主任医师,E-mail:ringring5452000@163.com。
更新日期/Last Update: 1900-01-01