Background: Numerous epidemiologic studies and experimental data support potential anti-tumor effects of green tea and its main component, epigallocatechin-3-gallate (EGCG), in breast cancer. However, there is limited data on the effects of tea catechins on breast cancer in human intervention trials. The purpose of this study is to evaluate tumor proteomic changes after short-term pre-surgical administration of an oral green tea extract, Polyphenon E (Poly E), in women with operable breast cancer using reverse phase protein array (RPPA).
Methods: This is a phase II single-arm open-label trial of oral Poly E 800 mg daily for 2-4 weeks in women with histologically-confirmed breast cancer on core biopsy who were scheduled for surgical resection. Formalin-fixed paraffin-embedded (FFPE) tumor tissue from the diagnostic core biopsy (pre-treatment) and surgical resection (post-treatment) were analyzed for expression of the Ki-67 proliferation index, estrogen receptor (ER), progesterone receptor (PR), and HER2 by immunohistochemistry (IHC). Protein was extracted for RPPA analysis of 161 proteins, including components of the PI3K/AKT and MAPK pathways. Women were matched by age, breast cancer stage, ER/PR/HER2 status, and time interval between breast biopsy and surgery to untreated historical controls. Paired t-test was used to calculate changes in protein markers before and after Poly E treatment and 2-sample t-test to compare biomarker changes in the treatment and no treatment groups. All statistical analyses were 2-sided and performed using SAS version 9.1.
Results: From Feb 2008 to Sept 2009, 25 women were enrolled and 21 were evaluable. Median age: 50 years (range, 33-71); White/Hispanic/Black (%): 44/48/7; Stage 0/I/II/III (%): 11/48/30/11; hormone receptor +/- (%): 85/15. Mean duration on Poly E was 20 days (range, 13-36). We demonstrated significant correlations between RPPA and IHC for Ki-67 (0.46, P<0.0001), ER (0.45, P=0.0017), PR (0.46, P=0.0014), and a trend for HER2 (0.28, P=0.0923). Poly E treatment did not cause a significant decrease in Ki-67 compared to untreated controls (mean absolute change, -0.5% vs. +2.6%, P=0.83). In the Poly E-treated group, the RPPA results showed significant modulation of apoptosis and PI3K/AKT pathway proteins (P<0.05). After Bonferroni correction to adjust for multiple comparisons (P<0.00031), 11 markers remained statistically significant comparing change from baseline with Poly E treatment: up-regulation of MEK1, JNK2, and p38-MAPK; down-regulation of CDK4, HER2-pY1248, MAPK-pT202-Y204, MIG-6, mTOR-pS2448, PRAS40-pT246, Src-pY416, and Scr-pY527. Compared to untreated controls, the Poly E group had significant changes from baseline in 10 proteins: up-regulation of IRS1, p38-MAPK, Notch1, and YAP; down-regulation of ERCC1, MIG-6, p90RSK-pT359-S363, PRAS40-pT246, Smad3, and Src-pY416 (P<0.05).
Conclusions: Short-term administration of Poly E did not significantly decrease proliferation in breast tumor tissue; however, our RPPA data suggests that Poly E may act on alternative signaling pathways. The changes we observed in CDK4, HER2, Src, MAPK, and JNK expression are consistent with preclinical studies of EGCG. This is one of the first human intervention trials to demonstrate the biologic effects of Poly E on growth factor signaling pathways in breast cancer.
Citation Format: Kimberly A. Ho, Davida Kornreich, James A. Cardelli, Jerry McLarty, Dawn L. Hershman, Matthew Maurer, Kevin Kalinsky, Bret Taback, Hanina Hibshoosh, Tao Su, Susan F. Refice, Katherine D. Crew. Evaluating tissue biomarker effects of an oral green tea extract, polyphenon E, using reverse phase protein array in women with operable breast cancer. [abstract]. In: Proceedings of the Eleventh Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2012 Oct 16-19; Anaheim, CA. Philadelphia (PA): AACR; Cancer Prev Res 2012;5(11 Suppl):Abstract nr A73.