Effect of Apollon siRNA combined with tetramethylpyrazine on prolife-ration and apoptosis of leukemia K562 cells

JIA Xiu-Hong; XIAO Fei-Fei; LI Jian-Chang

doi:10.7499/j.issn.1008-8830.2014.02.008

PDF(2452 KB)

Chinese Journal of Contemporary Pediatrics ›› 2014, Vol. 16 ›› Issue (2) : 135-140. DOI: 10.7499/j.issn.1008-8830.2014.02.008

TOPIC OF HEMATOLOGY & ONCOLOGY

Effect of Apollon siRNA combined with tetramethylpyrazine on prolife-ration and apoptosis of leukemia K562 cells

JIA Xiu-Hong, XIAO Fei-Fei, LI Jian-Chang

Author information +

History +

Abstract

Objective To investigate the effect of small interfering RNA (siRNA) silencing Apollon gene combined with tetramethylpyrazine (TMP) on the proliferation and apoptosis of human chronic myeloid leukemia cell line K562. Methods K562 cells were divided into blank control, negative control, and RNA interference (RNAi) group. For the RNAi group, the pGPHI-GFP-Neo-Apollon eukaryotic expression vector based on the best Apollon siRNA fragments screened out in previous experiments was constructed; the blank control group received no treatment, and the negative control group was transfected with negative plasmid vector. The mRNA and protein expression of Apollon was measured by RT-PCR and cell immunofluorescence, respectively. Additionally, TMP (320 μg/mL) was applied to set TMP, TMP+negative control, and TMP+RNAi groups. The cell viability and apoptosis rate were determined by MTT assay and flow cytometry, respectively. Results The constructed vector was stably expressed in K562 cells. The RNAi group had significantly lower mRNA and protein expression of Apollon than the blank control group and negative control (P<0.05). The RNAi group had significantly increased proliferation inhibition rate and apoptosis rate, as compared with the blank contorl group (P<0.05). The TMP+RNAi group had significantly increased proliferation inhibition rate and apoptosis rate, as compared with the RNAi, and TMP groups (P<0.05). Conclusions Apollon siRNA can significantly inhibit the proliferation and promote the apoptosis of K562 cells, and the addition of TMP can further increase the proliferation inhibition rate and apoptosis rate, suggesting that siRNA technology combined with drugs has a significant potential value in the treatment of leukemia.

Key words

Apollon gene / RNA interference / Tetramethylpyrazine / Leukemia K562 cell

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

JIA Xiu-Hong, XIAO Fei-Fei, LI Jian-Chang. Effect of Apollon siRNA combined with tetramethylpyrazine on prolife-ration and apoptosis of leukemia K562 cells[J]. Chinese Journal of Contemporary Pediatrics. 2014, 16(2): 135-140 https://doi.org/10.7499/j.issn.1008-8830.2014.02.008

References

[1] Long J, Parkin B, Ouillette P, et al. Multiple distinct molecular mechanisms influence sensitivity and resistance to MDM2 inhibitors in adult acute myelogenous leukemia[J]. Blood, 2010, 116(1): 71-80.
[2] Sung KW, Choi J, Hwang YK, et al. Overexpression of apollon, an antiapoptotic protein, is associated with poor prognosis in childhood de novo acute myeloid leukemia[J]. Clin Cancer Res, 2007, 13(17): 5109-5114.
[3] 张伟, 戴碧涛, 汪明宇, 等. 川芎嗪对白血病细胞黏附运动和侵袭的影响[J]. 重庆医科大学学报, 2008, 33(4): 403-406.
[4] Srinivasula SM, Ashwell JD. IAPs: what's in a Name?[J]. Mol Cell, 2008, 30(2): 123-135.
[5] Bartke T, Pohl C, Pvrowolakis G, et al. Dual role of BRUCE as an antiapoptotic IAP and a chemeric E2/E3 ubiquitin Ligase[J]. Mol Cell, 2004, 14(6): 801-811.
[6] Smolewski P, Robak T. Inhibitors of apoptosis proteins (IAPs) as potential molecular targets for therapy of hematological malignancies[J]. Curr Mol Med, 2011, 11(8): 633-649.
[7] Ismail EA, Mahmoud HM, Tawfik LM, et al. BIRC6/Apollon gene expression in childhood acute leukemia: impact on therapeutic response and prognosis[J]. Eur J Haematol, 2012, 88(2): 118-127.
[8] 陈建发, 李宇华, 陈引香, 等. Apollon siRNA 提高肝癌细胞化疗敏感性的实验研究[J]. 南方医科大学学报, 2011, 31(10): 1701-1704.
[9] Amundson SA, Myers TG, Scudiero DS, et al. An informatics approach identifying markers of chemosensitivity in human cancer cell lines[J]. Cancer Res, 2000, 60(21): 6101-6110．
[10] 杨岚, 杨平地, 梁蓉, 等. 川芎嗪联合环胞霉素A 逆转白血病多药耐药的研究[J]. 癌症, 2000, 19(4): 304-306.
[11] 李乃农, 陈元仲, 吕联皇, 等. 白血病细胞端粒酶活性的研究[J]. 白血病淋巴瘤, 2002, 11(5): 21-28.
[12] 吴涛, 瞿桂芳. 川芎嗪对白血病K562 细胞增殖和凋亡的影响[J]. 中国冶金工业医学杂志, 2010, 27(1): 5-7.
[13] Pennati M, Millo E, Gandellini P, et al. RNA interferencemediated validation of survivin and Apollon/BRUCE as new therapeutic targets for cancer therapy[J]. Curr Top Med Chem, 2012, 12(2): 69-78.