miR-216b-5p对喉癌TU686细胞增殖、迁移和侵袭的影响及其机制

doi:10.13481/j.1671-587X.20230115

Abstract

Abstract:

Objective To investigate the effect of miR-216b-5p on the proliferation， migration，invasion and apoptosis of laryngeal squamous cell cancer（LSCC） TU686 cells，and to analyze the target genes in laryngeal cancer cells and its possible mechanism. Methods The TU686 cells were divided into control group and miR-216b-5p group；the cells in control group were infected with the nonsense sequence through lentivirus，and the cells in miR-216b-5p group were infected with overexpressed miR-216b-5p by lentivirus.The expression levels of miR-216b-5p in the cells in two groups were detected by real-time fluorescence quantitative PCR（RT-qPCR） method， the clone formation rates of the cells in two groups were detected by clone formation assay，the scratch healing rates of cells in two groups were detected by cell scratch assay，the number of invasion cells in two groups was detected by Transwell assay，and the apoptotic rates of cells in two groups were detected by flow cytometry. The TargetScan website was used to predict the potential target genes of miR-216b-5p. The expression levels of target gene mRNA were detected by RT-qPCR method.The targeting relationship between miR-216b-5p and the target gene was detected by dual-luciferase assay. Results After lentivirus infection， compared with control group， the expression level of miR-216b-5p in the cells in miR-216b-5p group was significantly increased（P<0.01）.Compared with control group，the clone formation rate and the scratch healing rate of the cells in miR-216b-5p group were significantly decreased（P<0.01），the number of invasion cells was significantly decreased（P<0.01）， and the apoptotic rate was significantly increased（P<0.01）. Targetscan website predicted that autophagy related gene 5 （ATG5） was the potential target gene of miR-216b-5p；compared with control group，the expression level of ATG5 mRNA in the cells in miR-216b-5p group was significantly decreased（P<0.01）.The dual-luciferase assay proved that there was a targeting correlation between ATG5 and miR-216b-5p. Conclusion MiR-216b-5p can inhibit the proliferation， migration， and invasion and induce apoptosis of laryngeal cancer cells to exert a tumor suppressor effect， and its mechanism may be related to the targeted regulation of the expression level of ATG5.

Key words: Laryngeal neoplasms, Laryngeal squamous cell cancer, MiR-216b-5p, TU686 cells, Autophagy related gene 5

CLC Number:

R739.65

Zongnan YU,Ying CUI. Effect of miR-216b-5p on proliferation, migration and invasion of laryngeal cancer TU686 cells and its mechanism[J].Journal of Jilin University(Medicine Edition), 2023, 49(1): 116-121.

Figures/Tables 6

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References 20

1	MARKOU K， CHRISTOFORIDOU A， KARASMANIS I， et al. Laryngeal cancer： epidemiological data from Νorthern Greece and review of the literature［J］. Hippokratia， 2013， 17（4）： 313-318.
2	LI Y S， YAN B R， WANG X， et al. ALKBH5-mediated m6A modification of lncRNA KCNQ1OT1 triggers the development of LSCC via upregulation of HOXA9［J］. J Cell Mol Med， 2022， 26（2）： 385-398.
3	JENCKEL F， KNECHT R. State of the art in the treatment of laryngeal cancer［J］. Anticancer Res， 2013， 33（11）： 4701-4710.
4	LU T X， ROTHENBERG M E. microRNA［J］. J Allergy Clin Immunol， 2018， 141（4）： 1202-1207.
5	NILSEN T W. Mechanisms of microRNA-mediated gene regulation in animal cells［J］. Trends Genet， 2007， 23（5）： 243-249.
6	PILLAI R S， BHATTACHARYYA S N， FILIPOWICZ W. Repression of protein synthesis by miRNAs： how many mechanisms？［J］. Trends Cell Biol， 2007， 17（3）： 118-126.
7	FILIPOWICZ W， BHATTACHARYYA S N， SONENBERG N. Mechanisms of post-transcriptional regulation by microRNAs： are the answers in sight？［J］. Nat Rev Genet， 2008， 9（2）： 102-114.
8	TAN J， JING Y Y， HAN L， et al. microRNA-203 inhibits invasion and induces apoptosis of laryngeal cancer cells via targeting LASP1［J］. Eur Rev Med Pharmacol Sci， 2018， 22（19）： 6350-6357.
9	ZHOU M， WANG Y， ZHANG C M， et al. microRNA-195-5p suppresses the proliferation， migration， invasion and epithelial-mesenchymal transition of laryngeal cancer cells in vitro by targeting E2F3［J］. Exp Ther Med， 2021， 22（4）： 1078.
10	MENBARI M N， RAHIMI K， AHMADI A， et al. miR-216b-5p inhibits cell proliferation in human breast cancer by down-regulating HDAC8 expression［J］. Life Sci， 2019， 237： 116945.
11	YOU Y， TAN J X， GONG Y， et al. microRNA-216b-5p functions as a tumor-suppressive RNA by targeting TPT1 in pancreatic cancer cells［J］. J Cancer， 2017， 8（14）： 2854-2865.
12	刘浩，穆兰，黄志伟，等. 喉鳞状细胞癌相关microRNA的筛选及功能鉴定［J］. 锦州医科大学学报， 2021， 42（2）： 40-44.
13	LEE R C， FEINBAUM R L， AMBROS V. The C. elegans heterochronic gene Lin-4 encodes small RNAs with antisense complementarity to Lin-14［J］. Cell， 1993， 75（5）： 843-854.
14	GRIFFITHS-JONES S， GROCOCK R J， VAN DONGEN S， et al. miRBase： microRNA sequences， targets and gene nomenclature［J］. Nucleic Acids Res， 2006， 34（Database issue）： D140-D144.
15	REDA EL SAYED S， CRISTANTE J， GUYON L， et al. microRNA therapeutics in cancer： current advances and challenges［J］.Cancers （Basel），2021，13（11）： 2680.
16	LIU W， LI X L， TAN X， et al. microRNA-204-3p inhibits metastasis of pancreatic cancer via downregulating MGAT1［J］. J BUON， 2021， 26（5）： 2149-2156.
17	HE S Z， WANG Q. microRNA-548c-5p inhibits the proliferation of breast cancer cells through regulating Wnt/β-catenin signaling pathway［J］. Eur Rev Med Pharmacol Sci， 2020， 24（7）： 3795-3804.
18	LU J， GETZ G， MISKA E A， et al. microRNA expression profiles classify human cancers［J］. Nature， 2005， 435（7043）： 834-838.
19	TRANG P， MEDINA P P， WIGGINS J F， et al. Regression of murine lung tumors by the let-7 microRNA［J］. Oncogene， 2010， 29（11）： 1580-1587.
20	KOTA J， CHIVUKULA R R， O’DONNELL K A，et al. Therapeutic microRNA delivery suppresses tumorigenesis in a murine liver cancer model［J］. Cell， 2009， 137（6）： 1005-1017.

[1]	ZHAN Shihua, WANG Ping, YIN Wanzhong, ZHU Wei. Enhancement effect of resveratrol on sensitivity of laryngeal carcinoma Hep-2 cells to cisplatin and its mechanism [J]. Journal of Jilin University Medicine Edition, 2015, 41(02): 282-286.
[2]	WANG He-bin，WANG Ya-fang，SHEN Miao-yan，LI Na，ZHAO Li-jing，TENG Bo. Apoptosis of human laryngeal 　carcinoma Hep-2 cells induced by siRNA silencing eIF4E and its mechanism [J]. Journal of Jilin University Medicine Edition, 2014, 40(01): 39-43.
[3]	LI Guang-yu,JIANG Xiao-dan,MENG Cui-da,GAO Ge,ZHU Dong-dong. Expressions \|of \|SLeX and CD24 in laryngeal squamous cell carcinoma tissues [J]. J4, 2011, 37(2): 304-307.
[4]	CUI Xiang-Yan, HONG Xin, CHEN Wei-Lun, XU Gong, HUANG Ge-Xin, CHU Wei. Expression \|of Brg1 in laryngeal squamous cell carcinoma tissue and significance [J]. J4, 2010, 36(3): 546-549.
[5]	SONG Zhao-Xia, ZHU Wei, LI Peng, LIU Bing. Association between anti-oncogene PTEN single nucleotide polymorphism and laryngocarcinoma [J]. J4, 2009, 35(5): 918-922.
[6]	CHEN Wei-lun,LI Qiu-ming,ZHU Wei,GUO Xiao-feng，CUI Hang. Synchronous detection of cell cycle and apoptosis in laryngeal carcinoma cells and its significance [J]. J4, 2008, 34(5): 854-859.
[7]	ZHANG Lian-zhi, SUN Ji-feng, WANG Shao-hua,WANG Wei-fang, YANG Liu,WANG Ning， LIU Jian-kai. Effects of indomethacin on proliferation and iNOS activity of laryngeal cancer cell Hep-2 [J]. J4, 2008, 34(2): 251-253.
[8]	XU Cheng-bi，TENG Bo，LI Chang-qing，JIN Chun-shun. Promoter methylation of RASSF1A gene in laryngealsquamous cell carcinoma and protein expression [J]. J4, 2006, 32(2): 326-329.
[9]	XU Hai-yang,LU Xiu-ying,MA Jing,LI Xiao-ming. Expressions of TNFR1, TNFR2 and TRAF2 in laryngeal squamous carcinoma [J]. J4, 2006, 32(2): 322-325.
[10]	FU Zhong-ying,ZHANG Wei-tian,GUO Xiao-feng,QUAN Cheng-shi. Relationships between E-cadherin gene mutation,its abnormalprotein expression and metastasis of laryngeal cancer [J]. J4, 2005, 31(5): 788-790.
[11]	SHI Xiao-ru，LI Xiao-ming，WANG Zhe-wen，LU Xiu-ying. Expression of survivin gene in laryngeal carcinoma and its relationship with apoptosis [J]. J4, 2005, 31(5): 761-764.
[12]	FU Zhong-ying, QUAN Cheng-shi，GUO Xiao-feng. Expressions of CD44v6 protein and α- andβ-catenin mRNA in metastasis of laryngeal cancer [J]. J4, 2004, 30(6): 919-921.
[13]	XIN Ding，TENG Bo,LI Wei, XU Ya-hui, WU Dong-hui. Relationship between ICAM-1 expression andbiological characters of laryngeal cancer [J]. J4, 2004, 30(5): 687-689.
[14]	LI Ye，JIN Chun-shun，ZHAO Ming，FENG Hua，SUN Cui. Significance of proliferation and expansion of lymphatic capillaries in pericancerous tissue in lymphatic metastasisof laryngeal carcinoma [J]. J4, 2004, 30(3): 444-446.
[15]	DU Bo，WANG Ping，GUO Xiao-feng，DU Bao-dong. Expression of tissue-inhibitors of metalloproteinases-1and prognosis of laryngeal carcinoma [J]. J4, 2004, 30(1): 126-128.

Effect of miR-216b-5p on proliferation, migration and invasion of laryngeal cancer TU686 cells and its mechanism

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