Oncogenic functions of hMDMX in in vitro transformation of primary human fibroblasts and embryonic retinoblasts

Lenos, Kristiaan; de Lange, Job; Teunisse, Amina FAS; Lodder, Kirsten; Verlaan-de Vries, Matty; Wiercinska, Eliza; van der Burg, Marja JM; Szuhai, Karoly; Jochemsen, Aart G

doi:10.1186/1476-4598-10-111

Molecular Cancer

Table 3 Fold induction mRNA per cell line after Nutlin-3 treatment

From: Oncogenic functions of hMDMX in in vitro transformation of primary human fibroblasts and embryonic retinoblasts

Cell line	Nutlin-3 treatm (h)	*p53*			*p21*			*hMDM2*			*hMDM2-p2*			*hMDMX*			*PUMA*			*GADD45a*			*SURVIVIN*
VH10 wt	0	1.00	±	0.19	1.00	±	0.16	1.00	±	0.16	1.00	±	0.17	1.00	±	0.16	1.00	±	0.23	1.00	±	0.38	1.00	±	0.16
	6	1.10	±	0.18	9.36	±	1.34	28.62	±	5.40	43.93	±	6.93	1.38	±	0.21	4.90	±	1.06	5.17	±	1.45	0.90	±	0.14
	24	0.79	±	0.18	11.98	±	2.09	26.42	±	5.42	66.60	±	11.19	0.95	±	0.16	4.49	±	1.25	4.89	±	1.79	0.02	±	0.00
shp53	0	1.00	±	0.09	1.00	±	0.11	1.00	±	0.25	1.00	±	0.09	1.00	±	0.09	1.00	±	0.23	1.00	±	0.11	1.00	±	0.13
	6	0.85	±	0.17	2.24	±	0.24	1.56	±	0.31	4.26	±	0.43	0.91	±	0.17	1.84	±	0.35	1.46	±	0.28	1.18	±	0.14
	24	0.63	±	0.54	5.36	±	0.45	4.08	±	0.92	9.46	±	0.73	1.00	±	0.12	3.29	±	0.54	1.28	±	0.12	0.85	±	0.11
HRasV12-shp53	0	1.00	±	0.35	1.00	±	0.27	1.00	±	0.32	1.00	±	0.28	1.00	±	0.29	1.00	±	0.41	1.00	±	0.28	1.00	±	0.23
	6	0.81	±	0.24	1.34	±	0.33	1.29	±	0.39	1.97	±	0.56	1.09	±	0.41	1.19	±	0.40	1.14	±	0.30	0.99	±	0.22
	24	0.84	±	0.28	2.11	±	0.50	1.44	±	0.36	2.37	±	0.66	1.18	±	0.30	1.53	±	0.47	1.37	±	0.32	1.06	±	0.22
shRB-HRasV12-shp53	0	1.00	±	0.24	1.00	±	0.18	1.00	±	0.22	1.00	±	0.48	1.00	±	0.19	1.00	±	0.18	1.00	±	0.18	1.00	±	0.18
	6	0.88	±	0.24	1.54	±	0.30	1.16	±	0.30	2.61	±	0.97	1.22	±	0.24	1.22	±	0.54	1.49	±	0.63	1.00	±	0.20
	24	0.98	±	0.27	2.49	±	0.67	1.68	±	0.66	6.32	±	2.59	1.18	±	0.38	1.35	±	0.35	1.76	±	0.48	0.59	±	0.16
HA-hMDMX	0	1.00	±	0.17	1.00	±	0.22	1.00	±	0.03	1.00	±	0.08	1.00	±	0.34	1.00	±	0.14	1.00	±	0.09	1.00	±	0.58
	6	0.74	±	0.23	4.14	±	0.67	3.47	±	1.26	11.83	±	1.00	1.42	±	0.36	3.05	±	0.49	1.25	±	0.58	1.52	±	0.64
	24	1.63	±	0.25	6.86	±	1.71	11.92	±	1.36	18.14	±	2.03	1.80	±	0.58	5.45	±	1.00	2.88	±	0.93	0.48	±	0.20
HRasV12-HA-hMDMX	0	1.00	±	0.14	1.00	±	0.16	1.00	±	0.16	1.00	±	0.16	1.00	±	0.22	1.00	±	0.29	1.00	±	0.14	1.00	±	0.16
	6	0.90	±	0.16	6.78	±	1.16	9.01	±	1.83	17.33	±	2.97	1.09	±	0.22	3.09	±	0.91	1.27	±	0.21	0.79	±	0.22
	24	0.79	±	0.10	10.38	±	2.76	12.08	±	1.58	26.21	±	2.95	0.74	±	0.11	3.50	±	0.85	1.69	±	0.19	0.26	±	0.03
shRb-HRasV12-HA-hMDMX	0	1.00	±	0.10	1.00	±	0.05	1.00	±	0.09	1.00	±	0.07	1.00	±	0.05	1.00	±	0.26	1.00	±	0.05	1.00	±	0.06
	6	1.04	±	0.13	4.57	±	0.46	6.46	±	1.61	13.53	±	2.08	1.01	±	0.13	3.10	±	0.63	1.73	±	0.26	0.81	±	0.13
	24	0.99	±	0.14	5.76	±	0.92	6.55	±	0.94	15.36	±	2.04	0.96	±	0.14	5.33	±	1.27	1.71	±	0.28	0.23	±	0.03

VH10 cells were treated with 10 μM Nutlin-3 for the indicated times and analyzed using qRT-PCR. Expression was normalized for the housekeeping genes CAPNS1 and TBP. The induction of mRNA expression upon Nutlin-3 treatment per cell line is shown for p53, p21, hMDM2, hMDM2-p2, hMDMX, PUMA, GADD45alpha and SURVIVIN. For each cell line the basal mRNA expression is set at 1.0.

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ISSN: 1476-4598

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