Effect of epidermal growth factor receptor tyrosine kinase inhibition on epithelial proliferation in normal and premalignant breast.

Abstract

The factors controlling epithelial proliferation in ductal carcinoma in situ (DCIS) are unclear. Antiestrogens are effective in the prevention of the majority of estrogen receptor-positive, but not estrogen receptor (ER)-negative breast cancers, which suggests that other factor(s) are promoting proliferation in ER-negative DCIS. Mutated or overexpressed tyrosine kinases are frequently associated with tumor development. Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase that is involved with mitogenesis and is expressed in ER-negative DCIS. We hypothesized that EGFR is central in driving proliferation in ER-negative/EGFR-positive DCIS. The purpose of this study was to establish whether the EGFR tyrosine kinase inhibitor (EGFR-TKI), ZD1839 (Iressa), can reduce epithelial proliferation and increase apoptosis in EGFR-positive DCIS. Breast tissue from 16 women undergoing surgery for DCIS were implanted into 16-32 immunosuppressed mice/experiment (8 xenografts/mouse). Treatment commenced 2 weeks after implantation and consisted of once daily oral gavage with ZD1839 at doses ranging from 10 to 200 mg/kg for 14-28 days; appropriate controls were present. Xenografts were removed on days 14, 21, 28, and 42 after implantation and then assessed for proliferation (LI) by Ki67 immunostaining and apoptosis index (AI) by morphology. All Ps reported are two-sided. Overall, a 56% reduction in epithelial proliferation was seen with Iressa in EGFR-positive DCIS. EGFR-TK inhibition compared with vehicle controls resulted in a fall in Geometric Mean Labeling Index (LI) after 14 days (day 28) of treatment both in ER-negative/EGFR-positive DCIS [6.5% interquartile range (IQR, 3.8-11.1) versus 13.9% (IQR, 12.0-16.3%); F(1,3) = 103; P = 0.002] and ER-positive/EGFR-positive DCIS [4.6% (IQR, 3.9-5.2%) versus 11.7% (IQR, 9.2-15.5); F(1,2) = 32.3; P = 0.03]. EGFR-TK inhibition had similar effects on the "at risk" normal breast epithelium adjacent to DCIS in the treated epithelium LI day 28 [ZD1839 2.2% (IQR, 1.7-3.3%) compared with control 3.8% (IQR, 2.4-5.4%); F(1,14) = 29.2; P = 0.00009] and in addition increased epithelial apoptotic index at day 21 [ZD1839 0.38 (0.23-0.83) compared with control 0.19 (0.1-0.25); F(1,6) = 12.2; P = 0.013]. The effect on epithelial proliferation was still significant after 28 days of treatment [for both DCIS (F1,29) = 24; P = 0.039 and normal breast F(1,6) = 47.3; P = 0.0005]. EGFR-TK inhibition with ZD1839 offers a novel approach to the treatment of EGFR-positive DCIS, regardless of ER status, and provides a potential new chemopreventative approach in patients at high risk of breast cancer.