ShRNA-mediated gene silencing of β-catenin inhibits growth of human colon cancer cells

Table 1 Oligonucleotides sequences of β-catenin specific and negative control siRNA

Name	Sequence of siRNA	Target sites
Pgenesil-CAT1	AACAGTCTTACCTGGACTCTG	0290-0310
Pgenesil-CAT2	AAAGGCAATCCTGAGGAAGAG	0355-0375
Pgenesil-Neg	GACTTCATAAGGCGCATGC	No homology

Table 2 Oligonucleotides sequences of primer pairs

Goal gene	Upstream primer	Downstream primer	PCR frag(bp)
β-actin	5'-TCCTGTGGATCCACGAAACT-3'	5'-GAAGCATTTGCGGTGGACGAT-3'	330
β-catenin	5'-AGATGCAGCAACTAAACAGGA-3'	5'-GTACTACATTTTAAGCCATCT-3'	290
C-myc	5'-TACCCTCTCAACGACAGCAG-3'	5'-TCTTGACA TTCTCCTCGGTG-3'	477
CyclinD1	5'-GCCAACCTCCTCAACGACCGG-3'	5'-GTCCATGTTCTGCTGGGCCTG-3'	744

Table 3 The mRNA expression intensities of genes in different groups normalized by β-actin

Goal gene	Blank Control	Negative	CAT1	CAT2
β-catenin	0.98 ± 0.02	0.96 ± 0.03	0.51 ± 0.03	0.55 ± 0.01
C-myc	0.97 ± 0.01	0.96 ± 0.02	0.48 ± 0.04	0.52 ± 0.01
CyclinD1	0.98 ± 0.02	0.97 ± 0.03	0.47 ± 0.02	0.50 ± 0.02
P		aP > 0.05	aP > 0.05	aP > 0.05

^aP: Observation group vs blank control group.

Table 4 The western blot analysis for genes in different groups normalized by β-actin

Goal gene	Blank Control	Negative	CAT1	CAT2
β-catenin	0.95 ± 0.02	0.89 ± 0.04	0.52 ± 0.02	0.56 ± 0.03
C-myc	0.97 ± 0.01	0.92 ± 0.02	0.54 ± 0.01	0.54 ± 0.04
CyclinD1	0.94 ± 0.03	0.95 ± 0.01	0.53 ± 0.02	0.51 ± 0.01
P		aP > 0.05	aP < 0.05	aP < 0.05

^aP: Observation group vs blank control group.

Table 5 The Inhibition rate of cell proliferation in different group

	Blank Control (%)	Negative (%)	CAT1 (%)	CAT2 (%)
36 h	10	14	21	18
48 h	13	17	28	24
60 h	15	19	39	33
72 h	18	21c	54a	47a

^aP < 0.05 (vs Blank),

^cP > 0.05 (vs Blank).

Table 6 Effect of shRNA interference on cell cycle distribution and apoptosis

Group	G0/G1 (%)	S (%)	G2/M (%)	Apoptosis (%)
Blank	36.51 ± 2.52	47.32 ± 3.31	16.45 ± 1.52	0.78 ± 0.11
Negative	40.32 ± 2.74c	43.21 ± 2.12	16.57 ± 1.83	1.05 ± 0.15c
CAT1	81.71 ± 5.73a	14.34 ± 1.34	4.55 ± 0.35	5.21 ± 0.47a
CAT2	78.33 ± 5.32a	17.51 ± 1.81	5.31 ± 0.57	4.89 ± 0.38a

^aP < 0.05 (vs Blank);

^cP > 0.05 (vs Blank).