Phosphoinositide-3-kinase, catalytic, alpha polypeptide RNA interference inhibits growth of colon cancer cell SW948

doi:10.3748/wjg.v18.i26.3458

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World Journal of Gastroenterology

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1007-9327

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Baishideng Publishing Group Inc, 7041 Koll Center Parkway, Suite 160, Pleasanton, CA 94566, USA

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World J Gastroenterol. Jul 14, 2012; 18(26): 3458-3464
Published online Jul 14, 2012. doi: 10.3748/wjg.v18.i26.3458

Table 1 Oligonucleotides sequences of primer pairs

Goal gene	Upstream primer	Downstream primer	PCR frag (bp)
β-actin	5’-TCCTGTGGATCCACGAAACT-3’	5’-GAAGCATTTGCGGTGGACGAT-3’	330-bp
PIK3CA	5’-CCCTGCTCATCAACTAGGAAACC-3'	5’-TTGCCGTAAATCATCCCCATT-3'	160-bp
Akt	5’-GGACAACGCCATCCAGACT-3’	5’-GCCAGGGACACCTCCATCTC-3’	121-bp
c-myc	5’-TACCCTCTCAACGACAGCAG-3’	5’-TCTTGACA TTCTCCTCGGTG-3’	477-bp
cyclinD1	5’-GCCAACCTCCTCAACGACCGG-3'	5’-GTCCATGTTCTGCTGGGCCTG-3’	744-bp

PCR: Polymerase chain reaction; PIK3CA: Phosphoinositide-3-kinase, catalytic, alpha polypeptide.

Table 2 Levels of mRNA expression and protein expression of target genes in different groups normalized by β-actin

Goal gene	CA1	CA2	Negative	Blank
mRNA expression
PIK3CA	0.51 ± 0.04^a	0.49 ± 0.03^a	0.92 ± 0.01	0.93 ± 0.03
Akt	0.50 ± 0.03^b	0.48 ± 0.01^b	0.93 ± 0.04	0.92 ± 0.02
c-myc	0.49 ± 0.01^a	0.50 ± 0.04^a	0.90 ± 0.02	0.91 ± 0.03
cyclinD1	0.45 ± 0.02^a	0.51 ± 0.01^a	0.96 ± 0.03	0.98 ± 0.01
Protein expression
PIK3CA	0.53 ± 0.01^a	0.54 ± 0.02^a	0.92 ± 0.03	0.91 ± 0.02
Akt	0.49 ± 0.02^b	0.55 ± 0.03^b	0.94 ± 0.03	0.95 ± 0.04
c-myc	0.51 ± 0.03^b	0.52 ± 0.04^b	0.92 ± 0.02	0.95 ± 0.01
cyclinD1	0.54 ± 0.04^b	0.56 ± 0.03^b	0.93 ± 0.01	0.93 ± 0.03

PIK3CA: Phosphoinositide-3-kinase, catalytic, alpha polypeptide.

^aP = 0.001,

^bP = 0.000 vs negative or blank group.

Table 3 Phosphoinositide-3-kinase, catalytic, alpha polypeptide short hairpin RNA suppressed SW948 cell proliferation (%)

	Blank	Negative	CA1	CA2
36 h	13	14	17	15
48 h	14	17	29^a	25^a
60 h	16	19	38^a	34^a
72 h	17	20	53^b	48^b

^aP = 0.001,

^bP = 0.000 vs negative or blank group.

Table 4 Effect of short hairpin RNA interference on cell cycle distribution and apoptosis (%)

Group	G₀/G₁	S	G₂/M	Apoptosis
Blank	35.41 ± 2.63	47.54 ± 3.87	16.74 ± 1.59	0.86 ± 0.13
Negative	40.12 ± 2.54	41.71 ± 2.16	17.57 ± 1.26	0.95 ± 0.11
CA1	79.72 ± 4.63^a	15.24 ± 1.28^a	4.37 ± 0.43^b	5.11 ± 0.32^a
CA2	77.93 ± 4.31^a	16.71 ± 0.89^a	5.17 ± 0.29^a	4.73 ± 0.32^b

^aP = 0.001,

^bP = 0.000 vs negative or blank group.

Citation: Huang WS, Wang TB, He Y, Chen YJ, Zhong SL, Tan M. Phosphoinositide-3-kinase, catalytic, alpha polypeptide RNA interference inhibits growth of colon cancer cell SW948. World J Gastroenterol 2012; 18(26): 3458-3464
URL: https://www.wjgnet.com/1007-9327/full/v18/i26/3458.htm
DOI: https://dx.doi.org/10.3748/wjg.v18.i26.3458