|Table of Contents|

Screening of naked cuticle homolog 1 interacting proteins by yeast two-hybrid system

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

Issue:
2022 05
Page:
757-762
Research Field:
Publishing date:

Info

Title:
Screening of naked cuticle homolog 1 interacting proteins by yeast two-hybrid system
Author(s):
LIU Qian1; YU Huayi1; SHEN Ying1; WANG Wanyi2; WANG Linlin2; HAN Xiaodong2
1.Wujin Hospital Affiliated with Jiangsu University/The Wujin Clinical College of Xuzhou Medical University,Jiangsu Changzhou 213017,China;2.College of Life Sciences,Inner Mongolia Agriculture University,Inner Mongolia Hohhot 010018,China.
Keywords:
colon cancer; yeast two-hybrid; tumor markers; naked cuticle homolog 1; immunoprecipitation
PACS:
R735.3+5
DOI:
10.3969/j.issn.1672-4992.2022.05.001
Abstract:
Objective：To screen the naked cuticle homolog 1(NKD1) interacting proteins by yeast two-hybrid system,and clarify the biological function of NKD1.Methods:NKD1 gene was cloned from the cDNA of SW480 cells and successfully constructed into pGBKT7 bait vector.Yeast two-hybrid technology was used to screen from colon cancer SW480 cell library.Results:Thirteen positive clones were obtained.After sequencing and blast comparison in the database,eight different protein coding genes were identified.Rotation experiments further verified that these eight protein coding genes could activate the His3,Ade2 and LacZ reporter genes in yeast two-hybrid system.Four of them encode YWHAE protein and its homologous protein.In colon cancer HCT116 cells,NKD1 and YWHAE could bind to each other by immunoprecipitation assay.Conclusion:NKD1 can interact with YWHAE in colon cancer cells to regulate glucose intake by regulating insulin sensitivity.

References:

[1]BRENNER H,KLOOR M,POX CP.Colorectal cancer［J］.Lancet,2014,383(9927):1490-1502.
[2]CHEN WQ,ZHENG RS,BAADE PD,et al.Cancer statistics in China,2015［J］.CA:A Cancer Journal for Clinicians,2016,66(2):115-132.
[3]SIEGEL RL,MILLER KD,JEMAL A.Cancer statistics,2016［J］.CA:A Cancer Journal for Clinicians,2016,66(1)：7-30.
[4]MURALIDHAR S,FILIA A,NSENGIMANA J,et al.Vitamin D-VDR signaling inhibits Wnt/beta-catenin-mediated melanoma progression and promotes antitumor immunity［J］.Cancer Res,2019,79:5986-5998.
[5]HOFFMANN J,CORBETT L,DETJEN K,et al.The role of MAPK-signalling in pancreatic neuroendocrine cancer［J］.Neuroendocrinology,2017,10(5):8.
[6]MASUI K,HARACHI M,CAVENEE WK,et al.mTOR complex 2 is an integrator of cancer metabolism and epigenetics［J］.Cancer Letters,2020,478:1-7.
[7]LIU Q,LU W,YANG C,et al.HBXIP activates the PPARdelta/NF-kappaB feedback loop resulting in cell proliferation［J］.Oncotarget,2018,9(1):404-417.
[8]LIU T,YU T,HU H,et al.Knockdown of the long non-coding RNA HOTTIP inhibits colorectal cancer cell proliferation and migration and induces apoptosis by targeting SGK1［J］.Biomed Pharmacother,2018,98:286-296.
[9]ZENG W,WHARTON KA,MACK JA,et al.Naked cuticle encodes an inducible antagonist of Wnt signalling［J］.Nature,2000,403(6771):789-795.
[10]KATOH M.Molecular cloning,gene structure,and expression analyses of NKD1 and NKD2［J］.Int J Oncol,2001,19(5):963-969.
[11]MARSDEN AN,DERRY SW,SCHNEIDER I,et al.The Nkd EF-hand domain modulates divergent wnt signaling outputs in zebrafish［J］.Developmental Biology,2018,1(434):63-73.
[12]STANCIKOVA J,KRAUSOVA M,KOLAR M,et al.NKD1 marks intestinal and liver tumors linked to aberrant Wnt signaling［J］.Cell Signal,2015,27(2):245-256.
[13]ZHANG S,WANG Y,DAI SD,et al.Down-regulation of NKD1 increases the invasive potential of non-small-cell lung cancer and correlates with a poor prognosis［J］.BMC Cancer,2011,11(1):1-10.
[14]ZHANG S,LI J,YIN ZY,et al.Expression pattern and clinicopathologic significance of NKD1 in human primary hepatocellular carcinoma［J］.Acta Pathologica Microbiologica et Immunologica Scandinavica,2015,123(4):315-320.
[15]LYU ZD,ZHANG L,LIU XP,et al.NKD1 down-regulation is associated with poor prognosis in breast invasive ductal carcinoma［J］.Int J Clin Exp Pathol,2015,8(4):4015-4021.
[16]ANGONIN D,VAN RAAY TJ.Nkd1 functions as a passive antagonist of Wnt signaling［J］.PLoS One,2013,8(8):e74666.
[17]SEREBRIISKII I,ESTOJAK J,BERMAN M,et al.Approaches to detecting false positives in yeast two-hybrid systems ［J］.Biotechniques,2000,28(2):332-336.
[18]徐东刚.酵母双杂交系统的应用研究进展［J］.国外医学临床生物化学与检验学分册,2001,22(06)：281-282. XU DG.Research progress of yeast two hybrid system［J］.Foreign Medical Clinical Biochemistry and Laboratory Volume,2001,22(06)：281-282.
[19]LIU Q,DENG JZ,YANG CX,et al.DPEP1 promotes the proliferation of colon cancer cells via the DPEP1/MYC feedback loop regulation［J］.Biochemical and Biophysical Research Communications,2020,532(4):520-527.

Memo

Memo:
National Natural Science Foundation of China(No.81872275）；国家自然科学基金面上项目（编号：81872275）；江苏省卫生健康委科研基金面上项目（编号：M2020002）
Last Update: 2022-01-27