miR-219调控PRKCI表达在舌鳞状细胞癌中作用和机制

doi:10.11904/j.issn.1002-3070.2018.06.006

实用肿瘤学杂志 ›› 2018, Vol. 32 ›› Issue (6): 510-514.doi: 10.11904/j.issn.1002-3070.2018.06.006

miR-219调控PRKCI表达在舌鳞状细胞癌中作用和机制

王蕾, 姜寰宇, 宋凯镔

哈尔滨医科大学附属肿瘤医院头颈外科(哈尔滨 150081)

收稿日期:2018-07-08 出版日期:2018-12-28 发布日期:2018-12-27
通讯作者: 宋凯镔, E-mail:11714398@qq.com
作者简介:王蕾, 女, (1988-), 硕士, 住院医师, 从事头颈肿瘤学的研究。

miR-219 regulates PRKCI expression in tongue squamous cell carcinoma

WANG Lei, JIANG Huanyu, SONG Kaibin

Department of Head and Neck Surgery, Harbin Medical University Cancer Hospital, Harbin 150081, China

Received:2018-07-08 Online:2018-12-28 Published:2018-12-27

摘要/Abstract

摘要： 目的探讨PRKCI与miR-219表达的相关性以及对舌鳞状细胞癌的细胞增殖、侵袭和转移的影响。方法利用双荧光素酶报告基因实验对预测的靶基因进行验证, 用qRT-PCR和Western blot实验检测外源过表达miR-219对舌鳞状细胞癌细胞中转染的PRKCI基因和蛋白表达的影响。最后在过表达miR-219的稳转细胞系中再过表达PRKCI, 利用MTT法、细胞平板克隆实验、划痕实验和Transwell小室法分析PRKCI对miR-219抑制舌鳞状细胞癌细胞增殖、克隆形成能力、迁移能力和侵袭能力的逆转效果。通过qRT-PCR法检测舌鳞状细胞癌组织中PRKCI基因的表达情况并进一步分析PRKCI和miR-219之间的关系。结果通过生物信息学分析预测得到miR-219下游靶基因为PRKCI。双荧光素酶报告基因实验结果显示miR-219能够降低野生型PRKCI报告基因载体的荧光活性。此外qRT-PCR实验和Western blot实验也显示miR-219能够下调PRKCI在舌鳞状细胞癌细胞中的表达。MTT结果显示过表达PRKCI能够逆转miR-219对舌鳞状细胞癌细胞增殖活性的抑制效应, 进一步通过细胞平板克隆实验、划痕实验以及Transwell实验证明PRKCI过表达能够逆转miR-219对TSCC细胞增殖、侵袭和转移的抑制效应。结论 miR-219通过直接靶向PRKCI、负调控PRKCI的表达发挥抑制肿瘤的作用。在舌鳞状细胞癌组织中, miR-219与PRKCI的表达呈负相关。

关键词: 舌鳞状细胞癌, miR-219, PRKCI

Abstract: Objective The Objective of this study were to investigate the effects of miR-219 on cell proliferation, invasion and metastasis and the correlation between PRKCI and miR-219 expression in tongue squamous cell carcinoma. Methods The luciferase reporter gene assay was used to verify the predicted target gene.The expression of PRKCI in tongue squamous cell carcinoma cells overexpressed exogenous miR-219 was detected by qRT-PCR and Western blot.Finally, the reverse effects of PRKCI on the proliferation, clone formation, migration and invasion ability were examined in stable overexpressing miR-219 tongue squamous cell carcinoma(TSCC)cells by MTT assay, cell plate cloning assay, scratch assay and Transwell chamber assays.qRT-PCR assay was used to determine the expression of PRKCI gene and miR-219 in tongue squamous cell carcinoma tissues, and the relationship between PRKCI and miR-219 was further analyzed. Results The bioinformatics analysis predicted that the downstream target gene of miR-219 was PRKCI.The double luciferase reporter gene assay showed that miR-219 was able to reduce the fluorescence activity of the wild type PRKCI reporter vector.In addition, qRT-PCR and Western blot also showed that miR-219 could down-regulate the expression of PRKCI in TSCC cells.MTT results showed that overexpression of PRKCI could reverse the inhibitory effect of miR-219 on the proliferation of TSCC cells, and further demonstrated that the overexpression of PRKCI could reverse the inhibitory effect of miR-219 on the proliferation, invasion and metastasis of TSCC cells by cell plate cloning, scratch and Transwell experiments. Conclusion MiR-219 plays a role in inhibiting tumor by directly targeting PRKCI and negatively regulating the expression of PRKCI.miR-219 was negatively correlated with PRKCI expression in tongue squamous cell carcinoma.

Key words: Squamous cell carcinoma of the tongue, miR-219, PRKCI

中图分类号:

R739.86

王蕾, 姜寰宇, 宋凯镔. miR-219调控PRKCI表达在舌鳞状细胞癌中作用和机制[J]. 实用肿瘤学杂志, 2018, 32(6): 510-514.

WANG Lei, JIANG Huanyu, SONG Kaibin. miR-219 regulates PRKCI expression in tongue squamous cell carcinoma[J]. PRACTICAL ONCOLOGY JOURNAL, 2018, 32(6): 510-514.

参考文献

1 Sano D, Myers JN.Metastasis of squamous cell carcinoma of the oral tongue[J].Cancer Metastasis Rev, 2007, 26:645-662.
2 Liu X, Wang A, Heidbreder CE, et al.MicroRNA-24 targeting RNA-binding protein DND1 in tongue squamous cell carcinoma[J].FEBS Lett, 2010, 584:4115-4120.
3 Jia LF, Wei SB, Gan YH, et al.Expression, regulation and roles of miR-26a and MEG3 in tongue squamous cell carcinoma[J].Int J Cancer, 2014, 135(10):2282-2293.
4 Yang YL, Chu JY, Luo ML, et al.Amplification of PRKCI, located in 3q26, is associated with lymph node metastasis in esophageal squamous cell carcinoma[J].Genes Chromosomes Cancer, 2008, 47(2):127-136.
5 Regala RP, Weems C, Jamieson L, et al.Atypical protein kinase Ciota is an oncogene in human non-small cell lung cancer[J].Cancer Res, 2005, 65(19):8905-8911.
6 Justilien V, Walsh MP, Ali SA, et al.The PRKCI and SOX2 oncogenes are coamplified and cooperate to activate Hedgehog signaling in lung squamous cell carcinoma[J].Cancer Cell, 2014, 25(2):139-151.
7 Zhou B, Chen H, Wei D, et al.A novel miR-219-SMC4-JAK2/Stat3 regulatory pathway in human hepatocellular carcinoma[J].J Exp Clin Cancer Res, 2014, 33:55.
8 Friedman RC, Farh KK, Burge CB, et al.Most mammalian mRNAs are conserved targets of microRNAs[J].Genome Res, 2008, 19(1):92-105.
9 Fields AP, Regala RP.Protein kinase C iota:human oncogene, prognostic marker and therapeutic target[J].Pharmacol Res, 2007, 55(6):487-497.
10 Regala RP, Weems C, Jamieson L, et al.Atypical protein kinase Ciota is an oncogene in human non-small cell lung cancer[J].Cancer Res, 2005, 65(19):8905-8911.
11 Campa D, CanzianF, Kaaks R.Genetic variability in the PRKCI gene and prostate cancer risk[J].Cell Cycle, 2012, 11(2):209.
12 Yang YL, Chu JY, Luo ML, et al.Amplification of PRKCI, located in 3q26, is associated with lymph node metastasis in esophageal squamous cell carcinoma[J].Genes Chromosomes Cancer, 2008, 47(2):127-136.
13 Yang YL, Chu JY, Luo ML, et al.Amplification of PRKCI, located in 3q26, is associated with lymph node metastasis in esophageal squamous cell carcinoma[J].Genes Chromosomes Cancer, 2008, 47(2):127-136.
14 Frederick LA, Matthews JA, Jamieson L, et al.Matrix metalloproteinase-10 is a critical effector of protein kinase Ciota-Par6 alpha-mediated lung cancer[J].Oncogene, 2008, 27:4841-4853.
15 Justilien V, Fields AP.Ect2 links the PKCiota-Par6 alpha complex to Rac1 activation and cellular transformation[J].Oncogene, 2009, 28(41):3597-3607.
16 Regala RP, Weems C, Jamieson L, et al.Atypical protein kinase Ciota plays a critical role in human lung cancer cell growth and tumorigenicity[J].J Biol Chem, 2005, 280(35):31109-31115.
17 苑振楠, 宋凯镔.miR-219在舌鳞状细胞癌中的表达及意义研究[J].实用肿瘤学杂志, 2016, 30(5):420-424.
18 Justilien V, Walsh MP, Ali SA, et al.The PRKCI and SOX2 oncogenes are coamplifi ed and cooperate to activate Hedgehog signaling in lung squamous cell carcinoma[J].Cancer Cell, 2014, 25(2):139-151.

miR-219调控PRKCI表达在舌鳞状细胞癌中作用和机制

miR-219 regulates PRKCI expression in tongue squamous cell carcinoma

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 1

编辑推荐

Metrics

本文评价