Human colon carcinoma cell-line HCT116 transfected by antisense cDNA as a tool to study the Ku86 involvement in cell proliferation.

Abstract:

:In this work, we used colon carcinoma cell-line HCT116 to study the involvement of the 86-kDa subunit (Ku86) of DNA-protein kinase (DNA-PK) in human tumoural cell proliferation. We transfected these cells with a 639-bp cDNA encoding a Ku86 portion inserted into pcDNA3.1 vector in the antisense orientation. After selection by neomycin, we obtained more than 300 resistant colonies. In the Y'A5 colony that we chose as total population, we showed by PCR and RT-PCR that pcDNA3/Ku86 antisense was integrated in genomic DNA and that transcript was present. After cloning, we selected two clones, A20 and A23, which contained significatively reduced level of Ku86 protein. These two clones displayed a reduced DNA-PK activity from 44% to 71% and a slower growth than control cells. These results suggest that the HCT116 cell-line is a useful tool to investigate the role of Ku86 in the regulation of human tumoural cell growth.

journal_name

Cell Signal

journal_title

Cellular signalling

authors

Sadji Z,Le Romancer M,Lewin MJ,Reyl-Desmars F

doi

10.1016/s0898-6568(00)00126-1

subject

Has Abstract

pub_date

2000-12-01 00:00:00

pages

745-50

issue

11-12

eissn

0898-6568

issn

1873-3913

pii

S0898-6568(00)00126-1

journal_volume

12

pub_type

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