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Figure 5 | Breast Cancer Research

Figure 5

From: Autocrine IGF-I/insulin receptor axis compensates for inhibition of AKT in ER-positive breast cancer cells with resistance to estrogen deprivation

Figure 5

AZD5363-induced upregulation of IGF-IR, IGF-I and IGF-II is dependent on ER and FoxO3. A) LTED cells were treated with 10% DCC-FBS ± 2 µM AZD5363 for 24 hours. T47D cells were serum starved for 24 hours and then treated with 10% DCC-FBS ± 2 µM AZD5363 for 24 hours. RNA was extracted, reverse transcribed to cDNA and analyzed by real-time PCR. Threshold cycle values were normalized for actin. qPCR reactions with no input RNA were used as negative controls. Data are presented as fold versus control; each bar, mean ± SEM (n = 4; *P <0.05 versus Con, t-test). B) Mice bearing MCF-7 or ZR75-1 xenografts ≥150 mm3 were treated with vehicle or AZD5363 (150 mg/kg bid) for one day. Xenografts were harvested four hours after the last dose and homogenized. RNA was extracted, reverse transcribed to cDNA and analyzed by real-time PCR. Threshold cycle values were normalized for actin (MCF-7) or 36B4 (ZR75-1). Data are presented as fold versus control; each bar, mean ± SEM (n = 3; *P <0.05 versus Con, t-test). C, D) MCF-7/LTED cells were transfected with siRNA targeting FoxO3, ER, both (siboth) or a non-silencing control (siCon). Two days later the cells were treated with 10% DCC-FBS ± 2 µM AZD5363 for 24 hours. RNA was extracted and analyzed by real-time PCR as in A. Data are presented as fold versus control; each bar, mean ± SEM (n = 3). C) *P <0.01 versus siCon, t-test. D) *P <0.05 versus siCon; #P <0.05 versus AZD, t-test. DCC-FBS, dextran/charcoal-treated fetal bovine serum; ER, estrogen receptor; FoxO, forkhead box class O; LTED, long-term estrogen deprivation.

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