Effects of Wei Chang An regulated PTBP3 on expression of multiple genes in human gastric cancer cells

Abstract

Abstract: AIM: To study the effect of PTBP3 on the expression of Bcl-2,Stat3 in human gastric cancer cells, researching the related mechanisms between the anti-tumor effect of Wei Chang An(WCA) with regulated the expression of PTBP3 and other genes in gastric cancer cells.METHODS: Balb/c mice were planted MKN-45 cancer cells which were stably transfected with the Plasmids contained the siRNA of PTBP3 to establish experimental model, after that were randomizedly divided into WCA high dose group, WCA low dose group, 5-FU group and control group. The effects of PTBP3 and WCA on the tumor growth were observed.The mRNA and protein expression of PTBP3, Bcl-2 and Stat3 were detected with Utilized Real-time Quantitative PCR and Western Blot.RESULTS: In the silencing PTBP3 model and no-silencing PTBP3 model, the tumor weights of WCA high and low dose group and 5-FU group were lower than those in the control group in the same model, the differences were statistically significant (P values all are 0.000); the tumor weight of silencing PTBP3 model was lower than that in the no-silencing PTBP3 model, the difference was statistically significant (P=0.000). Real-time Quantitative PCR and Western Blot showed: compared with the no-silencing PTBP3 model, in the silencing PTBP3 model, the mRNA expression of PTBP3, Bcl-2 and Stat3 were down regulated, the difference was statistically significant, the protein expression of PTBP3, Bcl-2 and Stat3 were down regulated too.In the no-silencing PTBP3 model, compared with the control group, the mRNA expression of PTBP3, Bcl-2 and Stat3 in WCA groups and 5-FU group were down regulated, the difference was statistically significant, the protein expression of PTBP3, Bcl-2 and Stat3 were down regulated too. In the silencing PTBP3 model, there was no significant difference between WCA groups, 5-FU group with control group on the mRNA and protein expression of PTBP3; the mRNA expression of Bcl-2 and Stat3 in WCA groups were lower than those in the control group, the difference was statistically significant, the protein expression of Bcl-2 and Stat3 were down regulated too.CONCLUSION: PTBP3 can up-regulated the expression of Bcl-2 and Stat3 in gastric cells, the anti-tumor mechanism of WCA related with WCA down-regulated the expression of PTBP3 and Bcl-2,Stat3 genes in gastric cancer cells.

Key words: PTBP3, Stat3, Bcl-2, invigorating spleen, Wei Chang An

CLC Number:

R965.2

CHEN Bin, MU Xiao-yan, ZHAO Ai-guang, TAO Li, XU Yan. Effects of Wei Chang An regulated PTBP3 on expression of multiple genes in human gastric cancer cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2014, 19(5): 481-487.

References

[1] Zhao AG, Li T, You SF, et al. Effects of Wei Chang An on expression of multiple genes in human gastric cancer grafted onto nude mice[J]. World J Gastroenterol, 2008,14(5):693-700.
[2] Hanako Y, Kappei T, Yoshihide K, et al. Isolation of a mammalian homologue of a fission yeast differentiation regulator[J]. Mol Cell Bio, 1999,19(5): 3829-3841.
[3] Brazão TF, Demmers J, van IJcken W, et al. A new function of ROD1 in nonsense-mediated mRNA decay[J]. FEBS Lett,2012,586(8):1101-1010.
[4] Schmittgen TD, Zakrajsek BA, Mills AG, et al. Quantitative reverse transcription-polymerase chain reaction to study mRNA decay: comparison of endpoint and real-time methods[J]. Anal Biochem,2000,285(2):194-204.
[5] Paoletti X, Oba K, Burzykowski T, et al. Benefit of adjuvant chemotherapy for resectable gastric cancer: a meta-analysis[J]. JAMA, 2010,303(17):1729-1737.
[6] Jemal A, Siegel R, Xu J, et al. Cancer statistics, 2010[J]. CA Cancer J Clin, 2010,60:277-300.
[7] Ferlay J, Shin HR, Bray F, et al. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008[J].Int J Cancer, 2010,127(12):2893-2917.
[8] Jemal A, Bray F, Center MM, et al. Global cancer statistics[J].CA Cancer J Clin, 2011,61(2):69-90.
[9] Spellman R, Llorian M, Smith CW. Crossregulation and Functional Redundancy between the Splicing Regulator PTB and Its Paralogs nPTB and ROD1[J]. Mol Cell,2007,27(3):420-434.
[10] Maniatis T, Tasic B. Alternative pre-mRNA splicing and proteome expansion in metazoans[J]. Nature, 2002 ,418(6894):236-243.
[11] Hagen RM, Ladomery MR. Role of splice variants in the metastatic progression of prostate cancer[J].Biochemical Society Transactions,2012,40(4):870-874.
[12] Pal S, Gupta R, Davuluri RV. Alternative transcription and alternative splicing in cancer[J]. Pharmacol Ther, 2012,136(3): 283-294.
[13] Kim YJ, Kim HS. Alternative Splicing and Its Impact as a Cancer Diagnostic Marker[J]. Genomics Inf, 2012,10(2): 74-80.
[14] Gao G, Dou QP. G(1) phase-dependent expression of bcl-2 mRNA and protein correlates with chemoresistance of human cancer cells[J].Mol Pharmacol, 2000,58(5): 1001-1010.
[15] Capuzzi D, Santoro E, Hauck WW, et al. Fhit expression in gastric adenocarcinoma: correlation with disease stage and survival[J].Cancer, 2000,88(1): 24-34.
[16] Levy DE, Darnell JE Jr. Stats: transcriptional control and biological impact[J].Nat Rev Mol Cell Biol, 2002,3(9): 651-662.
[17] Gong W, Wang L, Yao JC, et al.Expression of activated signal transducer and activator of transcription 3 predicts expression of vascular endothelial growth factor in and angiogenic phenotype of human gastric cancer[J].Clin Cancer Res, 2005, 11(4): 1386-1393.