[1]匡剑韧,魏平.lncRNA LSINCT5靶向结合miR-29c调控乳头状甲状腺癌的增殖迁移[J].第三军医大学学报,2018,40(20):1851-1857.
 KUANG Jianren,WEI Ping.Long non-coding RNA LSINCT5 promotes proliferation and metastasis of papillary thyroid cancer by binding to miR-29c[J].J Third Mil Med Univ,2018,40(20):1851-1857.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第20期
页码:
1851-1857
栏目:
基础医学
出版日期:
2018-10-30

文章信息/Info

Title:
Long non-coding RNA LSINCT5 promotes proliferation and metastasis of papillary thyroid cancer by binding to miR-29c
作者:
匡剑韧魏平
陆军军医大学(第三军医大学)第一附属医院内分泌科
Author(s):
KUANG Jianren WEI Ping

Department of Endocrinology, First Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400038, China

关键词:
lncRNA LSINCT5乳头状甲状腺癌增殖迁移
Keywords:
long non-coding RNAs LSINCT5 papillary thyroid cancer proliferation migration  
分类号:
R394.13;R730.23;R736.1
文献标志码:
A
摘要:

目的    探讨lncRNA LSINCT5靶向结合miR-29c促乳头状甲状腺癌(papillary thyroid cancer,PTC)增殖和迁移的作用及机制。方法高通量转录芯片筛选PTC癌组织和癌旁组织差异表达lncRNAs。实时荧光定量PCR(qRT-PCR)检测32例来源于本院的PTC配对癌组织和癌旁组织样本(其中男性18例,女性14例,平均年龄57岁)和PTC细胞系中LSINCT5的表达水平。PTC细胞株TPC-1和KAT-5中转染siNC、siLSINCT5-1、siLSINCT5-2后,CCK-8和Transwell迁移实验测定转染各组的细胞增殖和转移水平。采用双荧光素酶报告基因实验验证LSINCT5与miR-29c的靶向结合。TPC-1细胞中转染Control(空质粒组)、LSINCT5、LSINCT5-Mut以及共转染LSINCT5和miR29c后,qRTPCR和Western blot检测ITGB1和Ecadherin表达水平。结果与配对的癌旁组织相比,LSINCT5在癌组织中表达显著升高(P<0.05)。LSINCT5在PTC细胞系中表达均高于人正常甲状腺细胞HT-ori3,差异有统计学意义(P<0.05)。与转染siNC相比,PTC细胞株TPC1和KAT-5在转染siLSINCT5-1、siLSINCT5-2后,TPC-1和KAT-5细胞的增殖和迁移能力均显著降低(P<0.05)。双荧光素酶报告基因和RIP证实LSINCT5能够与miR-29c特异性结合。与转染LSINCT5组相比,共转染LSINCT5和miR-29c 组中TPC-1细胞增殖和迁移能力均显著降低(P<0.05),ITGB1 mRNA和蛋白表达水平显著下调(P<0.05),E-cadherin 蛋白表达水平上升。结论    LSINCT5能够发挥miRNAs分子海绵的功能,靶向结合miR-29c而抑制miR-29c-ITGB1轴的抑癌作用,促进PTC的增殖和转移

Abstract:

ObjectiveTo evaluate the role of long noncoding RNA (lncRNA) LSINCT5 in promoting the proliferation and metastasis of papillary thyroid cancer (PTC) and explore the mechanism. MethodslncRNA highthroughput chip was employed to screen the differentially expressed lncRNAs between PTC tissues and the adjacent tissues. qRT-PCR was used to detect the expression level of LSINCT5 in paired PTC tissues and adjacent tissues from 32 patients (including 18 male and 14 female patients with a mean age of 57 years) and in PTC and normal human thyroid cell lines. Human PTC cell lines TPC-1 and KAT-5 were transfected with siLSINCT5-1 and siLSINCT5-2 for knocking down LSINCT5 expression, and the changes in cell proliferation and invasion were analyzed using CCK-8 assay and Transwell analysis. Dual luciferase reporter analysis and RIP were used to detect the interaction between LSINCT5 and miR-29c. The mRNA and protein expressions of ITGB1 and E-cadherin in TPC-1 cells co-transfected with LSINCT5 and miR-29c were detected using qRT-PCR and Western blotting. ResultsCompared with the adjacent tissues, PTC tissues showed a significantly increased LSINCT5 expression (P<0.05). LSINCT5 expression was also significantly elevated in PTC cell lines compared to normal human thyroid HT-ori3 cells (P<0.05). Compared with siNC transfection, transfection with siLSINCT5-1 and siLSINCT5-2 for LSINCT5 knockdown significantly suppressed the proliferation and migration of TPC-1 and AKT-5 cells (P<0.05). Dual luciferase reporter and RIP analyses confirmed that LSINCT5 could specifically bind to miR-29c. Compared with LSINCT5 transfection alone, co-transfection with LSINCT5 and mir-29c significantly inhibited the cell proliferation and migration (P<0.05), lowered ITGB1 mRNA and protein expressions (P<0.05), and increased E-cadherin protein expression in TPC-1 cells. ConclusionLSINCT5 plays the role of miRNA molecular sponge, and by competitively binding to miR-29c, LSINCT5 attenuates the tumor-inhibiting effect of miR-29c-ITGB1 axis to promote the proliferation and metastasis of PTC.

参考文献/References:

[1]TORRE L A, BRAY F, SIEGEL R L, et al. Global cancer statistics, 2012[J]. CA Cancer J Clin, 2015, 65(2): 87-108. DOI: 10.3322/caac.21262.
[2]LI Q, LI H, ZHANG L, et al. Identification of novel long noncoding RNA biomarkers for prognosis prediction of papillary thyroid cancer[J]. Oncotarge, 2017, 8(28): 46136-46144. DOI: 10.18632/oncotarget.17556.
[3]LIYANARACHCHI S, LI W, YAN P, et al. Genomewide expression screening discloses long noncoding RNAs involved in thyroid carcinogenesis[J]. J Clin Endocrinol Metab, 2016, 101(11): 4005-4013. DOI: 10.1210/jc.20161991.
[4]JENDRZEJEWSKI J, THOMAS A, LIYANARACHCHI S, et al. PTCSC3 is involved in papillary thyroid carcinoma development by modulating S100A4 gene expression[J]. J Clin Endocrinol Metab, 2015, 100(10): E1370-E1377. DOI: 10.1210/jc.20152247.
[5]WANG C, YAN G, ZHANG Y, et al. Long noncoding RNA MEG3 suppresses migration and invasion of thyroid carcinoma by targeting of Rac1[J]. Neoplasma, 2015, 62(4): 541-549. DOI: 10.4149/neo_2015_065.
[6]WANG Y, GU J, LIN X, et al. lncRNA BANCR promotes EMT in PTC via the Raf/MEK/ERK signaling pathway[J]. Oncol Lett, 2018, 15(4): 5865-5870. DOI: 10.3892/ol.2018.8017.
[7]DING S, QU W, JIAO Y, et al. LncRNA SNHG12 promotes the proliferation and metastasis of papillary thyroid carcinoma cells through regulating wnt/betacatenin signaling pathway[J]. Cancer Biomark, 2018, 22(2): 217-226. DOI: 10.3233/CBM170777.
[8]ZHU H, LV Z, AN C, et al. OncolncRNA HOTAIR and its functional genetic variants in papillary thyroid carcinoma[J]. Sci Rep, 2016, 6: 31969. DOI: 10.1038/srep31969.
[9]JEONG S, LEE J, KIM D, et al. Relationship of focally amplified long noncoding on chromosome 1 (FAL1) lncRNA with E2F transcription factors in thyroid cancer[J]. Medicine, 2016, 95(4): e2592. DOI: 10.1097/MD.0000000000002592.
[10]ZHOU Q, CHEN J, FENG J, et al. Long noncoding RNA PVT1 modulates thyroid cancer cell proliferation by recruiting EZH2 and regulating thyroidstimulating hormone receptor (TSHR) [J]. Tumour Biol, 2016, 37(3): 3105-3113. DOI: 10.1007/s1327701541499.
[11]CHEN C, ZHOU L, WANG H, et al. Long noncoding RNA CNALPTC1 promotes cell proliferation and migration of papillary thyroid cancer via sponging miR30 family[J]. Am J Cancer Res, 2018, 8(1): 192-206.
[12]WANG X, LU X, GENG Z, et al. LncRNA PTCSC3/miR5745p governs cell proliferation and migration of papillary thyroid carcinoma via Wnt/betacatenin signaling[J]. J Cell Biochem, 2017, 118(12): 4745-4752. DOI: 10.1002/jcb.26142.

更新日期/Last Update: 2018-10-31