Figure 4. p21 knockdown impairs the oncolytic effect of the vaccinia virus. (A) Tumor-killing effect of TK-deleted vaccinia virus on MDA-MB-231 shp21 cells was reduced compared to that on MDA-MB-231 shCtrl cells. (B) Relative mRNA level of TK in MDA-MB-231 shp21 cells was lower than that in MDA-MB-231 shCtrl cells. (C) Tumor-killing effect of wild-type vaccinia virus on MDA-MB-231 shp21 was greater than that on MDA-MB-231 shCtrl cells. (D) Replication ability of TK-deleted vaccinia virus in MDA-MB-231 shCtrl cells and MDA-MB-231 shp21 cells. (E) Photographs showing that TK-deleted VV-EGFP successfully infected MDA-MB-23 shCtrl and MDA-MB-231 shp21cells. (F) p21 KD in MDA-MB-231 cells inhibited the oncolytic effect of vaccinia virus in mouse tumor burden experiments. MDA-MB-231 shp21-derived cells (5 × 10⁶ per mouse) and MDA-MB-23 shCtrl-derived cells (8 × 10⁶ per mouse) were injected into the mice subcutaneously. Once the tumor reached approximately 500 mm³, 1 × 10⁶ VV-EGFP virus/100 μL in PBS or 100 μL PBS was injected into the tumor of the mice according to the group. Student’s t-test was used to compare two groups, and two-way analysis of variance was used to compare multiple groups. Data are shown as the mean ± standard deviation. shCtrl = short hairpin RNA control; shp21 = short hairpin RNA targeting p21; TK = thymidine kinase; VV-WT = vaccinia virus wild-type; NS = not significant; EGFP = enhanced green fluorescent protein; VV-EGFP= vaccinia virus carrying enhanced green fluorescent protein; PBS = phosphate-buffered saline.

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Breast Cancer Cells and Impairs the Tumor-Killing Effect of the Vaccinia Virus

Mariam Nafea, Aamna Basheer, Ohud Alkhaibry, Alanoud Alsaedan

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