Apoptotic Effects of psiRNA-STAT3 on 4T1 Breast Cancer Cells in Vitro

  • Zhou, Yue (School of Pharmacy, The Second Clinical Hospital) ;
  • Tian, Lin (School of Pharmacy, The Second Clinical Hospital) ;
  • Zhang, Ying-Chao (Department of Breast Surgery, The Second Clinical Hospital) ;
  • Guo, Bao-Feng (Department of Plastic Surgery, the China-Japan Union Hospital) ;
  • Zhou, Qing-Wei (Department of Biology and Medical Engineering, Institute of Regenerative Medicine, Jilin University)
  • Published : 2014.08.30


Background: The aim of this study was to investigate the effect of a Lipofectamine2000 (Life2000) Transfection Reagent transfected psiRNA-STAT3 plasmid on 4T1 breast cancer cells. Materials and Methods: MTT was used to detect the cell proliferation of breast cancer 4T1 cells at different periods (0h, 6h, 8h, 10h); the cell cycle was assessed by flow cytometry; variation of apoptosis and mitochondrial membrane potential was observed under a fluorescence microscope; immunohistochemical staining was used to determine the expression of caspase-3 and cyclin-D1 protein. Results: An obvious effect of inhibition to 4T1 cancer cells could be observed at 8h after the psiRNA-STAT3 was transfected. Typical alterations of apoptotic morphological features were visible in the psiRNA-STAT3 treatment group. Mitochondrial membrane potential decreased significantly, the number of cells was increased in G0/G1 phase, and the number of cells was decreased in S phase, and the data were statistically significant (p<0.05), compared with the Scramble and Mock groups. Expression of caspase-3 protein was increased significantly, while that of cyclin D1 was significantly decreased. Conclusions: Life2000 transfected psiRNA-STAT3 plasmid can inhibit 4T1 tumor cell proliferation and promote apoptosis of 4T1 tumor cells, which process depends on the regulation of expression of cyclin D1 and caspase-3 protein.


psiRNA-STAT3 plasmid;breast cancer 4T1 cells;apoptosis;protein expression


  1. Aggarwal B, Sethi.GA.K, wang.S, et al (2006). Targeting signaltransducer- and-activator-of-transcription-3 for prevention and therapy of cancer: modern target but ancient solution. Ann N Y Acad Sci, 1091, 151-69.
  2. Canzian F, Cox DG, Setiawan VW (2010). Comprehensive analysis of common genetic variation in 61 genes related to steroid hormone and insulin-like growth factor-I metabolism and breast cancer risk in the NCI breast and prostate cancer cohort consortium. Hum Mol Genet, 19, 3873-84.
  3. Chatter J, Kishore M, van den Akker F, Stark GR (2000). Association of STATs with relatives and friends. Trend Cell Biology, 10, 106-11.
  4. Chatter J, Kishore M, van den Akker F, Stark GR (2000). Association of STATs with relatives and friends. Trend Cell Biology, 10, 106-11.
  5. Chen B, Pan H, Zhu L, et al (2005). Progesterone Inhibits the estrogen-induced phosphoinositide 3-kinasegAKTgGSK- $3{\beta}gCyclin$ D1gpRB pathway to block uterine epithelial cell proliferation. Molec Endocrinol, 19, 1978-90.
  6. Chiba T, Yamada M, Aiso S (2009). Targeting the JAK2/STAT3 axisia Alzheimer's disease. Expert Opin Ther Targets, 13, 1155-67.
  7. DaoTong Liu, Peng Zhao, JingYan Han, et al (2014). Clinical and prognostic significance of SOX11 in breast cancer. Asian Pac J Cancer Prev, 15, 5483-6.
  8. Desagher S, Martinou JC (2000). Mitochondria as the central control point of apoptosis. Trends Cell Biol, 10, 369-77.
  9. Hsieh FC, Cheng G, Lin J (2005). Evaluation of potential Stat3- regulated genes in human breast cancer. Biochem Biophys Res Com, 335, 292-99.
  10. Grebenova D, Kuzelova K, Smetana K, et al (2003). Mitochondrial and en-doplasmic reticulum stress-induced apoptotic pathways are activat-ed by 5-aminolevulinic acid-based photodynamic therapy in HL60 leukemia cells. J Photochem Photobiol B, 69, 71-85.
  11. HishikawaK, Nakaki T, Fujii T (2000). Connective tissue growth factor Induces apoptosis via caspase-3 incultured human aortic smooth musclecens. Eur J Pharmacol, 392, 19-22.
  12. Kaufmann SH, Lee SH, Meng XW, et al (2008). Apoptosis is associated Caspase activation assays. Methods, 44, 262-72.
  13. Kollmar O, Rupertus K, Scheuer C, et al (2010). CXCR4 and CXCR7 regulate angiogenesis and CT26 WT tumor growth independent from SDF-1. Int J Cancer, 126, 1302-15.
  14. Ling Zhang, Lifang Gao, Lijuan Zhao, et al (2007). Intratumoral delivery and suppression of prostate tumor growth by attenuated Salmonella enterica serovar typhimurium carrying plasmid-based small interfering RNAs. Cancer Res, 67, 5859-64.
  15. Madoux F, Koenig M, Nelson E, et al (2010). Modulators of STAT Transcription factors for the targeted therapyof cancer ( STAT3 Activators) . Bethesda ( MD) : National Center for Biotechnology Information, 16, 2.
  16. Renl PJ, Sierra A, Juan A, et al (2002). Resistance to chemotherapy via STAT3 dependent over expression of Bcl- 2 in motastatic breast cancer cell. Oncogene, 21, 7611-18
  17. Reynoso-Noveron N, Mohar-Betancour TA (2013). Inflammatory Breast Cancer. London: Springer-Verlag, London, 11-3
  18. Shi ZB, Zhao D, Huang YY, et al (2012). Discovery, synthesis, andevaluationof small-moleculesignal transducer andactivator of transcription3 inhibitors. Chem Pharm Bull, 60, 1574-80.
  19. Tkach M, Rosemblit C, Rivas MA, et al (2013). p42/p44MAPK mediated Stat3Ser727 phosphorylation is required for progestin-induced full activation of Stat3 and breast cancer growth. Endocr Relat Cancer, 20, 197-201.
  20. Wang Q, Li J, Zheng S, et al (2012). Breast cancer stage at diagnosis and area-based socioeconomic status: a multicenter 10-year retrospective clinical epidemiological study in China. BMC Cancer, 12, 122-5.
  21. Wen Z, Yajie W (2007). Research progress of STAT3 and breast cancer. Mod Oncol, 15, 439-41.
  22. Xue Zhao; Xian Sun; Xiao-Li Li (2012). Expression and clinical significance of STAT3, P-STAT3, and VEGF-C in small cell lung cancer. Asian Pac J Cancer Prev, 13, 2873-7.
  23. Zhou Y, Guo BF, Zhang L, et al (2012). Gene therapy in a mouse tumor xenograft model of breast cancer by siRNA-mediated down-regulation of STAT3. Gene Ther Mol Biol, 14, 9-15.

Cited by

  1. Bevacizumab Regulates Cancer Cell Migration by Activation of STAT3 vol.16, pp.15, 2015,
  2. Inhibition of prostate cancer RM1 cell growth in vitro by hydroxyapatite nanoparticle-delivered short hairpin RNAs against Stat3 vol.16, pp.1, 2017,
  3. vol.33, pp.7, 2015,