|Table of Contents|

shRNA silencing CNTN1 gene expression inhibits proliferation and clone formation of breast cancer MDA-MB-468 cells

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2021 04
Page:
564-568
Research Field:
Publishing date:

Info

Title:
shRNA silencing CNTN1 gene expression inhibits proliferation and clone formation of breast cancer MDA-MB-468 cells
Author(s):
HE Sai1; GENG Jie2; YANG Xiaomin1; HOU Yanni1; FAN Yongguo1; ZHAO Jing1; ZHANG Jingyuan1; CHEN Nan1
1.Department of Breast Oncology,Shaanxi Provincial Cancer Hospital,Shaanxi Xi'an 710061,China;2.Department of Cardiovascular Internal Medicine,the Second Affiliated Hospital of Xi'an Jiaotong University,Shaanxi Xi'an 710004,China.
Keywords:
breast cancer; MDA-MB-468; CNTN1; proliferation; clone formation
PACS:
R737.9
DOI:
10.3969/j.issn.1672-4992.2021.04.005
Abstract:
Objective：To investigate the effect of short hairpin RNA (shRNA) targeted silencing of CNTN1 gene on proliferation and clone formation of breast cancer MDA-MB-468 cells.Methods:Real-time quantitative PCR (RT-PCR) and Western blot were used to detect the mRNA and protein levels of CNTN1 in MCF7-ADR,MDA-MB-468、MCF7 and Hs578T cells．MDA-MB-468 cells were randomly transfected with the successfully constructed shRNA vector fragments targeting CNTN1 gene (silencing group) or nonsense shRNA fragments (control group) via LipofectamineTM 2000.CNTN1 mRNA and protein expression was detected after 48 h transfection by RT-PCR and Western blot.The proliferation and clone formation abilities of cells were detected by MTT,clone formation assay and flow cytometry,respectively.Results:The relative expression level of CNTN1 mRNA and protein in MDA-MB-468 cells had the highest expression significantly (P＜0.05).MDA-MB-468 cells in transfection group underwent G1 phase block.The capabilities of proliferation,clone formation of MDA-MB-468 cells in transfection group decreased significantly compared with the non-transfection group and mock-vehicle group(P＜0.05).Conclusion:CNTN1 gene is highly expressed in breast cancer MDA-MB-468 cells,and its silencing expression may inhibit proliferation and clone formation．

References:

［1］ RACHEL CH,JESSICA H.Breast cancer knowledge and decisions made for contralateral prophylactic mastectomy:A survey of surgeons and women in the general population［J］.Plastic & Reconstructive Surgery,2019,143(5):936e-945e.
[2] LEE J.A Meta-analysis of the association between physical activity and breast cancer mortality［J］.Cancer Nursing,2019,42:252-261.
[3] LI ZX,CHONG SY,GULIMIRAN A,et al.Effect of sulindac derivative K-80003 in combination with MEK inhibitor cobimetinib in breast cancer cells［J］.Chinese Pharmacological Bulletin,2019,35(2):251-254.
[4] HAENISCH C,DIEKMANN H,KLINGER M,et al.The neuronal growth and regeneration associated Cntn1 (F3/F11/Contactin) gene is duplicated in fish:Expression during development and retinal axon regeneration［J］.Molecular and Cellular Neuroscience,2005,28(2):361-374.
[5] HE M,WU W.Contactin 1(CNTN1) expression associates with regional lymph node metastasis and is a novel predictor of prognosis in patients with oral squamous cell carcinoma［J］.Molecular Medicine Reports,2012,6(2):265-270.
[6] DOPPLER K,SCHUSTER Y,APPELTSHAUSER L,et al.Anti-CNTN1 IgG3 induces acute conduction block and motor deficits in a passive transfer rat model［J］.Journal of Neuroinflammation,2019,16(1):250-255.
[7] OLAKOGLU G,BERGSTROM-TYRBERG U,BERGLUND EO,et al.Contactin-1 regulates myelination and nodal/paranodal domain organization in the central nervous system［J］.Proc Natl Acad Sci,2014,111:E394-403.
[8] 陈楠,贺赛,丁艳妮,等.CNTN1基因对乳腺癌细胞株Hs578T增殖、迁移及侵袭能力的影响［J］.现代肿瘤医学,2016,24(19):3017-3020. CHEN N,HE S,DING YN,et al.Effects of recombinant plasmid of pEGFP-N1-CNTN1 on proliferation,clone formation,migration and invasion of Hs578T cells［J］.Modern Oncology,2016,24(19):3017-3020.
[9] SPAN PN.Survivin is an independent prognostic marker for risk stratification of breast cancer patients［J］.Clinical Chemistry,2020,50(11):1986-1993.
[10] STONE JK,KIM JH,VUKADIN L,et al.Hypoxia induces cancer cell-specific chromatin interactions and increases MALAT1 expression in breast cancer cells［J］.Journal of Biological Chemistry,2019,11(11):294-300.
[11] RVILLION F,HORNEZ L,PEYRAT JP.Quantification of c-erbB-2 gene expression in breast cancer by competitive RT-PCR［J］.Clinical Chemistry,2020,43(11):2114-2120.
[12] RAMIREZ JM,FEHM T,ORSINI M,et al.Prognostic relevance of viable circulating tumor cells detected by EPISPOT in metastatic breast cancer patients［J］.Clinical Chemistry,2020,60(1):214-221.
[13] REN YX,XIAO RD,LOU XM,et al.Research advance and application in the gene therapy of gene editing technologies［J］.Hereditas,2019,41(1):18-27.
[14] LEI JT,GOU X,ELLIS MJ.Abstract PD8-03:ESR1 gene fusions drive endocrine therapy resistance and metastasis in breast cancer［J］.Mol Cell Oncol,2018,5(6):e1526005.
[15] SRINIVAS PR,SRIVASTAVA S,HANASH S,et al.Proteomics in early detection of cancer［J］.Clinical Chemistry,2020,47(10):1901-1911.
[16] ISABEL LLLA.ARTHUR ASBURY LECTURE:Chronic inflammatory demyelinating polyradiculoneuropathy:Clinical aspects and new animal models of auto-immunity to nodal components［J］.Journal of the Peripheral Nervous System,2017,22(4):418-424.
[17] MARTIN POOT.Intragenic CNTNAP2 deletions:A bridge too far［J］.Molecular Syndromology,2017,8(3):118.
[18] CATRONA A,GERDING WM,CHRISTOPH F,et al.PLP1 and CNTN1 gene variation modulates the microstructure of human white matter in the corpus callosum［J］.Brain Structure and Function,2018,13(3):251-256.
[19] ERIK O BERGLUND,BARBARA RANSCHT.Molecular cloning and in situ localization of the human contactin gene (CNTN1) on chromosome 12q11-q12［J］.Genomics,1994,21(3):580-582.
[20] CHEN DH.Roles of contactin-1 in solid tumors［J］.World Chinese Journal of Digestology,2015,23(30):4785.
[21] SU JL,YANG PC,SHIH JY,et al.The VEGF-C/Flt-4 axis promotes invasion and metastasis of cancer cells［J］.Cancer Cell,2006,9:209-223.
[22] LAMPRIANOU S,CHATZOPOULOU E,THOMAS J,et al.Complex between contactin-1 and the protein tyrosine phosphatase PTPRZ controls the development of oligodendrocyte precursor cells［J］.Proceedings of the National Academy of Sciences of the United States of America,2011,108(42):17498.
[23] YAN J,WONG N,HUNG C,et al.Contactin-1 reduces E-cadherin expression via activating AKT in lung cancer［J］.PLoS One,2019,8:e65463.
[24] YU JW,WU SH,LU RQ,et al.Expression and significances of contactin-1 in human gastric cancer［J］.Gastroenterol Res Pract,2013:210205.
[25] BENSON JR,JATOI I.The global breast cancer burden［J］.Future Oncol,2012,8:697-702.
[26] CHEN N,HE S,GENG J,et al.Overexpression of Contactin 1 promotes growth,migration and invasion in Hs578T breast cancer cells［J］.BMC Cell Biology,2018,19:5-13.

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陕西省自然科学基础研究计划（编号：2020JM-680）
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