Objective To construct dual-regulated oncolytic adenoviruses carrying TAp63 gene and detect their antitumor effect on colon cancer stem cells.Methods Colon cancer stem cells were screened by flow cytometry.New recombinant adenoviruses were constructed according to the Adeasy system.The viral DNA acted as a template for PCR, then agarose gel electrophoresis assay was applied to confirm the viral DNA.CCK8 assay and cell apoptosis assay were used to detect the effects of recombinant adenoviruses on the proliferation of colon cancer stem cells and human normal liver cells, as well as on the apoptosis of colon cancer stem cells.Results The proportion of CD133/CD44 double positive cells was (42.47±0.81)% in HCT116 cells (P<0.05).The recombinant adenoviruses Arsz-TAp63 were successfully constructed without wild-type adenovirus contamination, and were shown to inhibit the proliferation of colon cancer stem cells HCT116 CD133/CD44, but not human normal liver cells L-02, and promote the apoptosis of HCT116 CD133/CD44 cells.Conclusion The recombinant adenoviruses Arsz-TAp63 selectively inhibit the proliferation and induce the apoptosis of HCT116 CD133/CD44 cells.
LUO Qifeng
,
DU Min
,
DAI Zhenling
,
FU Zhengwei
,
ZHANG Zhenyu
,
Shiva BASNET
,
GE Haiyan
. Dual-regulated oncolytic adenoviruses carrying TAp63 geneinhibit the growth of colon cancer stem cells[J]. Surgical Research and New Technique, 2018
, 7(2)
: 77
-82
.
DOI: 10.3969/j.issn.2095-378X.2018.02.001
[1] Chen W, Zheng R, Baade PD, et al.Cancer statistics in China,2015[J].CA Cancer J Clin, 2016, 66(2):115-132.
[2] Duarte S,Momier D, Baque P, et al.Preventive cancer stem cell-based vaccination reduces liver metastasis development in a rat colon carcinoma syngeneic model[J].Stem Cells, 2013, 31(3):423-432.
[3] Gerger A, Zhang W, Yang D, et al.Common cancer stem cell gene variants predict colon cancer recurrence[J].Clin Cancer Res, 2011, 17(21):6934-6943.
[4] Dhawan A, Kohandel M, Hill R, et al.Tumour control probability in cancer stem cells hypothesis[J].PLoS One, 2014, 9(5):e96093.
[5] Liu XY, Gu JF, Shi WF.Targeting gene-virotherapy for cancer[J].Acta Biochim Biophys Sin (Shanghai), 2005, 37(9):581-587.
[6] Zhang KJ, Wang YG, Cao X, et al.Potent antitumor effect of interleukin-24 gene in the survivin promoter and retinoblastoma double-regulated oncolytic adenovirus[J].Hum Gene Ther, 2009, 20(8):818-830.
[7] Luo Q, Liu H, Zhang Z, et al.A dual-regulated oncolytic adenovirus carrying TAp63 gene exerts potent antitumor effect on colorectal cancer cells[J].Am J Transl Res, 2017, 9(6):2966-2974.
[8] Jing F, Kim HJ, Kim CH, et al.Colon cancer stem cell markers CD44 and CD133 in patients with colorectal cancer and synchronous hepatic metastases[J].Int J Oncol, 2015, 46(4):1582-1588.
[9] Bellizzi A, Sebastian S, Ceglia P, et al.Co-expression of CD133(+)/CD44(+) in human colon cancer and liver metastasis[J].J Cell Physiol, 2013, 228(2):408-415.
