Effect of Skp2 antisense oligodeoxynucleotide on growth and proliferation of colorectal cancer SW 480 cells
10.3969/j.issn.1671-8348.2014.03.019
- VernacularTitle:Skp2反义寡核苷酸对大肠癌细胞SW480生长和增殖的影响
- Author:
Hongwei ZHOU
;
Jijian WANG
- Publication Type:Journal Article
- Keywords:
colorectal cancer;
antisense oligodeoxynucleotide;
Skp2;
P27kip1
- From:
Chongqing Medicine
2014;(3):307-310
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of S-phase kinase-associated protein 2 antisense oligodeoxynucleotide (Skp2 ASODN) on the growth and proliferation of colorectal cancer cells SW480 and its mechanism .Methods The liposome-mediated dif-ferent concentrations of Skp2 ASODN was adopted to transfect SW480 cells ,the final concentrations of 0 .050 ,0 .125 ,0 .250 ,0 .500 , 1 .000 ,2 .000 ,4 .000 μmol/L were respectively set up with Skp2 nonsense oligodeoxynucleotides(NSODN) and blank group as con-trol .Then the inhibited effect of growth and proliferation of colorectal cancer cells were measured by the inverted microscope obser-vationandmethylthiazolyltetrazolium(MTT),thecellcyclewassurveyedbytheflowcytometry(FCM).TheexpressionofmRNA and protein of Skp2 and P27kip1 were inspected by reverse transcription-polymerase chain reaction(RT-PCR) and immunocytochem-istry methods .Results The inverted microscope observed that the SW480 cells had no change in form and grew at a slower speed . When the Skp2 ASODN concentration reached 0 .125 μmol/L ,the inhibition rate was 14 .48% (P<0 .05) .As the Skp2 ASODN concentration increase ,its inhibition action on the growth and proliferation of SW480 cells was enhanced in a dose-dependent man-ner(P<0 .01) .The SW480 cells were increased in G0/G1 phase(P<0 .01) and decreased in S phase(P<0 .05) ,the progression of cell cycle was blocked .RT-PCR and the immunocytochemistry showed that both Skp2 mRNA and protein levels were down-regula-ted(P<0 .05) .Although the P27kip1 mRNA level had no change(P>0 .05) ,but its protein expression was elevates obviously (P<0 .01) .Conclusion Skp2 ASODN can inhibit the growth and proliferation of SW480 cells .Its possible mechanism is that the ubiq-uitin degradation of the Skp2 to P27kip1 is decreased by the gene occlusion effect of Skp2 ASODN ,thus the cell cycle progression is arrested .
