Simultaneous silencing of β‑catenin and signal transducer and activator of transcription 3 synergistically induces apoptosis and inhibits cell proliferation in HepG2 liver cancer cells

Wang,Xin‑Hong; Liu,Ming‑Na; Huang,Ping; Xu,Jun; Liu,Ai‑Yun; Chen,Jing; Lv,Cheng‑Qian; Xu,Rui‑Ling

doi:10.3892/mmr.2015.3595

Simultaneous silencing of β‑catenin and signal transducer and activator of transcription 3 synergistically induces apoptosis and inhibits cell proliferation in HepG2 liver cancer cells

Authors:
- Xin‑Hong Wang
- Ming‑Na Liu
- Ping Huang
- Jun Xu
- Ai‑Yun Liu
- Jing Chen
- Cheng‑Qian Lv
- Rui‑Ling Xu
View Affiliations

Affiliations: Department of Gastroenterology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China, Department of Gastroenterology, The First Hospital of Harbin, Harbin, Heilongjiang 150000, P.R. China
Published online on: April 3, 2015 https://doi.org/10.3892/mmr.2015.3595
Pages: 2263-2268

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The tumorigenesis and maintenance of a cancer cells is dependent upon the collaboration of multiple signaling pathways. Signal transducer and activator of transcription 3 (STAT3) and β‑catenin are at the center of multiple cancer‑associated signaling pathways; therefore, simultaneously targeting STAT3 and β‑catenin may be a potential cancer treatment, leading to induced lethality of cancer cells. In the present study, HepG2 liver cancer cells were transfected with small interfering RNA (siRNA) against β‑catenin and STAT3 alone or in combination. The cell growth was assessed using an MTT assay and the levels of cell apoptosis were detected using flow cytometry. Protein levels of caspase‑3, cleaved caspase‑3, poly(ADP‑ribose) polymerase (PARP) and cleaved PARP were determined using western blot analysis. Following siRNA transfection, β‑catenin and STAT3 protein levels decreased at 72 h. HepG2 cell growth inhibition and early apoptosis in the β‑catenin and STAT3 siRNA co‑transfection group were significantly greater than those in the groups transfected with β‑catenin or STAT3 siRNA alone. Decreased caspase‑3 and PARP levels, as well as enhanced cleavage of caspase‑3 and PARP were observed in the β‑catenin and STAT3 co‑transfection group. Simultaneous silencing of β‑catenin and STAT3 using siRNAs resulted in an enhanced loss of cell viability and induction of apoptosis in HepG2 liver cancer cells, suggesting that these genes are promising targets for the further preclinical and clinical development of anti‑cancer therapeutic strategies, which target several cancer signaling pathways simultaneously.

View Figures

View References

1	Yamakado K and Kudo M: Treatment strategies of intermediate-stage hepatocellular carcinomas in Japan (Barcelona Clinic Liver Cancer stage B). Oncology. 87(Suppl 1): 78–81. 2014. View Article : Google Scholar : PubMed/NCBI
2	Chan LH, Luk ST and Ma S: Turning hepatic cancer stem cells inside out - a deeper understanding through multiple perspectives. Mol Cells. Feb 4–2015.Epub ahead of print. View Article : Google Scholar
3	Wang XH, Sun X, Meng XW, et al: β-catenin siRNA regulation of apoptosis- and angiogenesis-related gene expression in hepatocellular carcinoma cells: potential uses for gene therapy. Oncol Rep. 24:1093–1099. 2010.PubMed/NCBI
4	Zulehner G, Mikula M, Schneller D, et al: Nuclear beta-catenin induces an early liver progenitor phenotype in hepatocellular carcinoma and promotes tumor recurrence. Am J Pathol. 176:472–481. 2010. View Article : Google Scholar :
5	Li W, Tong H, Huang X, et al: High levels of β-catenin promote IFNγ-induced apoptosis in hepatocellular carcinoma cells. Oncol Lett. 4:1092–1096. 2012.PubMed/NCBI
6	Guturi KK, Mandal T, Chatterjee A, et al: Mechanism of β-catenin-mediated transcriptional regulation of epidermal growth factor receptor expression in glycogen synthase kinase 3 β-inactivated prostate cancer cells. J Biol Chem. 287:18287–18296. 2012. View Article : Google Scholar : PubMed/NCBI
7	Wang XH, Meng XW, Sun X, et al: Wnt/β-catenin signaling regulates MAPK and Akt1 expression and growth of hepatocellular carcinoma cells. Neoplasma. 58:239–244. 2011. View Article : Google Scholar
8	Tian XH, Hou WJ, Fang Y, et al: XAV939, a tankyrase 1 inhibitior, promotes cell apoptosis in neuroblastoma cell lines by inhibiting Wnt/β-catenin signaling pathway. J Exp Clin Cancer Res. 32:1002013. View Article : Google Scholar
9	Liu YZ, Wu K, Huang J, et al: The PTEN/PI3K/Akt and Wnt/β-catenin signaling pathways are involved in the inhibitory effect of resveratrol on human colon cancer cell proliferation. Int J Oncol. 45:104–112. 2014.PubMed/NCBI
10	Fletcher S, Turkson J and Gunning PT: Molecular approaches towards the inhibition of the signal transducer and activator of transcription 3 (Stat3) protein. ChemMedChem. 3:1159–1168. 2008. View Article : Google Scholar : PubMed/NCBI
11	Morikawa T, Baba Y, Yamauchi M, et al: STAT3 expression, molecular features, inflammation patterns and prognosis in a database of 724 colorectal cancers. Clin Cancer Res. 17:1452–1462. 2011. View Article : Google Scholar : PubMed/NCBI
12	Park KW, Kundu J, Chae IG, Kim DH, Yu MH, Kundu JK and Chun KS: Carnosol induces apoptosis through generation of ROS and inactivation of STAT3 signaling in human colon cancer HCT116 cells. Int J Oncol. 44:1309–1315. 2014.PubMed/NCBI
13	Song X, Wang J, Zheng T, et al: LBH589 Inhibits proliferation and metastasis of hepatocellular carcinoma via inhibition of gankyrin/STAT3/Akt pathway. Mol Cancer. 12:1142013. View Article : Google Scholar : PubMed/NCBI
14	Tsang CM, Cheung YC, Lui VW, et al: Berberine suppresses tumorigenicity and growth of nasopharyngeal carcinoma cells by inhibiting STAT3 activation induced by tumor associated fibroblasts. BMC Cancer. 13:6192013. View Article : Google Scholar
15	Li W, Huang X, Tong H, et al: Comparison of the regulation of β-catenin signaling by type I, type II and type III interferons in hepatocellular carcinoma cells. PLoS One. 7:e470402012. View Article : Google Scholar
16	Zhang Y, Du XL, Wang CJ, et al: Reciprocal activation between PLK1 and Stat3 contributes to survival and proliferation of esophageal cancer cells. Gastroenterology. 142:521–530.e3. 2012. View Article : Google Scholar
17	Yang F, Jove V, Buettner R, et al: Sorafenib inhibits endogenous and IL-6/S1P induced JAK2-STAT3 signaling in human neuroblastoma, associated with growth suppression and apoptosis. Cancer Biol Ther. 13:534–541. 2012. View Article : Google Scholar : PubMed/NCBI
18	Wang XH, Liu BR, Qu B, et al: Silencing STAT3 may inhibit cell growth through regulating signaling pathway, telomerase, cell cycle, apoptosis and angiogenesis in hepatocellular carcinoma: potential uses for gene therapy. Neoplasma. 58:158–171. 2011. View Article : Google Scholar : PubMed/NCBI
19	Ogiwara H, Ui A, Shiotani B, et al: Curcumin suppresses multiple DNA damage response pathways and has potency as a sensitizer to PARP inhibitor. Carcinogenesis. 34:2486–2497. 2013. View Article : Google Scholar : PubMed/NCBI
20	Yuan K, Sun Y, Zhou T, McDonald J and Chen Y: PARP-1 regulates resistance of pancreatic cancer to TRAIL therapy. Clin Cancer Res. 19:4750–4759. 2013. View Article : Google Scholar : PubMed/NCBI
21	Booth L, Cruickshanks N, Ridder T, et al: PARP and CHK inhibitors interact to cause DNA damage and cell death in mammary carcinoma cells. Cancer Biol Ther. 14:458–465. 2013. View Article : Google Scholar : PubMed/NCBI
22	Dabir S, Kluge A, McColl K, et al: PIAS3 activates the intrinsic apoptotic pathway in non-small cell lung cancer cells independent of p53 status. Int J Cancer. 134:1045–1054. 2014. View Article : Google Scholar :
23	Swiatek-Machado K, Mieczkowski J, Ellert-Miklaszewska A, et al: Novel small molecular inhibitors disrupt the JAK/STAT3 and FAK signaling pathways and exhibit a potent antitumor activity in glioma cells. Cancer Biol Ther. 13:657–670. 2012. View Article : Google Scholar : PubMed/NCBI
24	Tierney BJ, McCann GA, Cohn DE, et al: HO-3867, a STAT3 inhibitor induces apoptosis by inactivation of STAT3 activity in BRCA1-mutated ovarian cancer cells. Cancer Biol Ther. 13:766–775. 2012. View Article : Google Scholar : PubMed/NCBI
25	Lee NJ, Choi DY, Song JK, et al: Deficiency of C-C chemokine receptor 5 suppresses tumor development via inactivation of NF-κB and inhibition of monocyte chemoattractant protein-1 in urethane-induced lung tumor model. Carcinogenesis. 33:2520–2528. 2012. View Article : Google Scholar : PubMed/NCBI
26	Bill MA, Nicholas C, Mace TA, et al: Structurally modified curcumin analogs inhibit STAT3 phosphorylation and promote apoptosis of human renal cell carcinoma and melanoma cell lines. PLoS One. 7:e407242012. View Article : Google Scholar : PubMed/NCBI
27	Xue M, Ge Y, Zhang J, et al: Fucoidan inhibited 4T1 mouse breast cancer cell growth in vivo and in vitro via downregulation of Wnt/β-catenin signaling. Nutr Cancer. 65:460–468. 2013. View Article : Google Scholar
28	Kundu J, Wahab SM, Kundu JK, et al: Tob1 induces apoptosis and inhibits proliferation, migration and invasion of gastric cancer cells by activating Smad4 and inhibiting β-catenin signaling. Int J Oncol. 41:839–848. 2012.PubMed/NCBI
29	Jaeger A, Baake J, Weiss DG and Kriehuber R: Glycogen synthase kinase-3beta regulates differentiation-induced apoptosis of human neural progenitor cells. Int J Dev Neurosci. 31:61–68. 2013. View Article : Google Scholar
30	Wang Y, Tao ZZ, Chen SM, et al: Application of combination of short hairpin RNA segments for silencing VEGF, TERT and Bcl-xl expression in laryngeal squamous carcinoma. Cancer Biol Ther. 7:896–901. 2008. View Article : Google Scholar : PubMed/NCBI