miR-217调控lncRNA&nbsp;MALAT1影响食管鳞癌细胞的生物学行为

徐健; 江跃全; 蔡华荣; 尹哲; 张奇

doi:10.13418/j.issn.1001-165x.2018.04.011

中国临床解剖学杂志 ›› 2018, Vol. 36 ›› Issue (4) : 408-413. DOI: 10.13418/j.issn.1001-165x.2018.04.011

实验研究

miR-217调控lncRNA MALAT1影响食管鳞癌细胞的生物学行为

徐健，　江跃全，　蔡华荣，　尹哲，　张奇

作者信息 +

miR-217 inhibits proliferation, migration, and invasion of esophageal squamous cell carcinoma cells by regulating lncRNA MALAT1

XU Jian, JIANG Yue-quan, CAI Hua-rong, YIN Zhe, ZHANG Qi

Author information +

文章历史 +

摘要

目的　探讨miR-217通过调控lncRNA MALAT1抑制食管鳞状细胞癌细胞的增殖，迁移和侵袭行为及其机制。方法　qPCR检测miR-217在食管鳞状细胞癌组织和不同细胞株中的表达情况；双荧光素酶报告基因检测miR-217与MALAT1之间的相互作用；MTT增殖实验检测抑制miR-217后食管鳞状细胞癌细胞的增殖能力的变化情况；划痕愈合试验和Transwell侵袭实验检测抑制miR-217后食管鳞状细胞癌细胞的迁移和侵袭行为的变化情况；Western blotting实验检测miR-217对MALAT1下游相关蛋白表达情况的影响。结果　与正常食管组织相比，食管鳞状细胞癌组织中miR-217的表达水平相对上调，与其他细胞株相比，Ec109细胞中miR-217表达最高；双荧光素酶实验证实miR-217能与MALAT1的3’ UTR特异性结合，可以调控MALAT1的表达与活性；抑制miR-217的表达后可以抑制食管鳞状细胞癌细胞的增殖、迁移和侵袭能力；抑制miR-217后MALAT1下游MIA2，ROBO1表达明显下调。结论　miR-217可以调控MALAT1的表达影响食管鳞状细胞癌细胞的生物学行为。

Abstract

Objective　To investigate the inhibitory effect of miR-217 on proliferation, migration and invasion of esophageal squamous cell carcinoma cells by regulating lncRNA MALAT1.　Methods　qPCR was used to detect the expression of miR-217 in esophageal squamous cell carcinoma and different cell lines. Dual luciferase reporter assay detected the interaction between miR-217 and MALAT1. MTT assay was used to detect the proliferation of esophageal squamous cell carcinoma cells after miR-217 was inhibited. Scratch healing assay and Transwell invasion assay were used to detect the migration and invasion of esophageal squamous carcinoma cells after miR-217 was inhibited. The effect of miR-217 on the expression of MALAT1 downstream protein was detected by Western blotting.　Results　Compared with normal esophageal tissue, the expression of miR-217 in esophageal squamous cell carcinoma was relatively up-regulated, and the expression of miR-217 was the highest in Ec109 cells compared with other cell lines. Dual luciferase assay confirmed that miR-217 could specifically bind to 3'UTR of MALAT1 and regulate the expression and activity of MALAT1. Inhibition of miR-217 expression could inhibit esophageal squamous cell carcinoma cell proliferation, migration and invasion ability. Down-regulation of miR-217 after MALAT1 MIA2, ROBO1 expression was significantly down-regulated.　Conclusion　miR-217 can regulate the expression of MALAT1 and affect the biological behavior of esophageal squamous cell carcinoma cells.

导出引用

徐健，　江跃全，　蔡华荣，　尹哲，　张奇. miR-217调控lncRNA MALAT1影响食管鳞癌细胞的生物学行为[J]. 中国临床解剖学杂志. 2018, 36(4): 408-413 https://doi.org/10.13418/j.issn.1001-165x.2018.04.011

XU Jian, JIANG Yue-quan, CAI Hua-rong, YIN Zhe, ZHANG Qi. miR-217 inhibits proliferation, migration, and invasion of esophageal squamous cell carcinoma cells by regulating lncRNA MALAT1[J]. Chinese Journal of Clinical Anatomy. 2018, 36(4): 408-413 https://doi.org/10.13418/j.issn.1001-165x.2018.04.011

参考文献

[1] Lin DC, Hao JJ, Nagata Y, et al. Genomic and molecular characterization of esophageal squamous cell carcinoma[J]. Nat Genet, 2014, 46(5): 467-473.
[2] Prabhu A, Obi KO, Rubenstein JH. The synergistic effects of alcohol and tobacco consumption on the risk of esophageal squamous cell carcinoma: a meta-analysis[J]. Am J Gastroenterol, 2014, 109(6): 822-827.
[3] Zeng W, Li H, Chen Y, et al. Survivin activates NF?κB p65 via the IKKβ promoter in esophageal squamous cell carcinoma[J]. Mol Med Rep, 2016, 13(2): 1869-1880.
\[4] Giri S, Pathak R, Aryal MR, et al. Incidence trend of esophageal squamous cell carcinoma: an analysis of surveillance epidemiology, and end results (SEER) database[J]. Cancer Causes Control, 2015, 26(1): 159-161.
[5] Wu C, Wang Z, Song X, et al. Joint analysis of three genome-wide association studies of esophageal squamous cell carcinoma in Chinese populations[J]. Nat Genet, 2014, 46(9): 1001-1006.
[6] Chen FJ, Sun M, Li SQ, et al. Upregulation of the long non-coding RNA HOTAIR promotes esophageal squamous cell carcinoma metastasis and poor prognosis[J]. Mol Carcinog, 2013, 52(11): 908-915.
[7] Yu C, Xue J, Zhu W, et al. Warburg meets non-coding RNAs: the emerging role of ncRNA in regulating the glucose metabolism of cancer cells[J]. Tumor Biol, 2015, 36(1): 81-94.
[8] Liu Y, Zhao L, Li D, et al. Microvesicle-delivery miR-150 promotes tumorigenesis by up-regulating VEGF, and the neutralization of miR-150 attenuate tumor development[J]. Protein Cell, 2013, 4(12): 932-941.
[9] Liu Y, Zhang M, Liang L, et al. Over-expression of lncRNA DANCR is associated with advanced tumor progression and poor prognosis in patients with colorectal cancer[J]. Int J Clin Exp Pathol, 2015, 8(9): 11480-11489.
[10] Inamura K. Major tumor suppressor and oncogenic non-coding RNAs: clinical relevance in lung cancer[J]. Cells, 2017, 6(2): 12-19.
[11] Tee AE, Liu B, Song R, et al. The long noncoding RNA MALAT1 promotes tumor-driven angiogenesis by up-regulating pro-angiogenic gene expression[J]. Oncotarget, 2016, 7(8): 8663-8675.
[12] Dong Y, Liang G, Yuan B, et al. MALAT1 promotes the proliferation and metastasis of osteosarcoma cells by activating the PI3K/Akt pathway[J]. Tumor Biol, 2015, 36(3): 1477-1486.
[13] Ren S, Liu Y, Xu W, et al. Long noncoding RNA MALAT-1 is a new potential therapeutic target for castration resistant prostate cancer[J]. J Urol, 2013, 190(6): 2278-2287.　　　　　[14] Han Y, Wu Z, Wu T, et al. Tumor-suppressive function of long noncoding RNA MALAT1 in glioma cells by downregulation of MMP2 and inactivation of ERK/MAPK signaling[J]. Cell Death Dis, 2016, 7(3): e2123.
[15] Ren D, Li H, Li R, et al. Novel insight into MALAT-1 in cancer: therapeutic targets and clinical applications[J]. Oncol Lett, 2016, 11(3): 1621-1630.
[16] Wu Y, Huang C, Meng X, et al. Long noncoding RNA MALAT1: insights into its biogenesis and implications in human disease[J]. Curr Pharm Des, 2015, 21(34): 5017-5028.
[17] Zhang B, Mao YS, Diermeier SD, et al. Identification and characterization of a class of MALAT1-like genomic loci[J]. Cell Rep, 2017, 19(8): 1723-1738.
[18] Ma XY, Wang JH, Wang JL, et al. Malat1 as an evolutionarily conserved lncRNA, plays a positive role in regulating proliferation and maintaining undifferentiated status of early-stage hematopoietic cells[J]. BMC Genomics, 2015, 16(1): 676-685.
[19] Leti F, Legendre C, Still CD, et al. Altered expression of MALAT1 lncRNA in nonalcoholic steatohepatitis fibrosis regulates CXCL5 in hepatic stellate cells[J]. Trans Res, 2017, 190: 25-39. e21.
[20] Lu L, Luo F, Liu Y, et al. Posttranscriptional silencing of the lncRNA MALAT1 by miR-217 inhibits the epithelial–mesenchymal transition via enhancer of zeste homolog 2 in the malignant transformation of HBE cells induced by cigarette smoke extract[J]. Toxicol Appl Pharmacol, 2015, 289(2): 276-285.
[21] Wang X, Li M, Wang Z, et al. Silencing of long noncoding RNA MALAT1 by miR-101 and miR-217 inhibits proliferation, migration, and invasion of esophageal squamous cell carcinoma cells[J]. J Biol Chem, 2015, 290(7): 3925-3935.
[22] Xiao H, Tang K, Liu P, et al. LncRNA MALAT1 functions as a competing endogenous RNA to regulate ZEB2 expression by sponging miR-200s in clear cell kidney carcinoma[J]. Oncotarget, 2015, 6(35): 38005-38009.
[23] Fesler A, Ju J. microRNA-based novel therapeutic development in gastrointestinal cancer[M]//Therapeutic Targets for Inflammation and Cancer: Novel Therapies for Digestive Diseases. 2017: 365-386.