Role of enolase-1 in response to hypoxia in breast cancer: Exploring the mechanisms of action

Gao,Jie; Zhao,Rongrong; Xue,Yan; Niu,Zuoxing; Cui,Kai; Yu,Fachang; Zhang,Bo; Li,Sheng

doi:10.3892/or.2013.2269

Role of enolase-1 in response to hypoxia in breast cancer: Exploring the mechanisms of action

Authors:
- Jie Gao
- Rongrong Zhao
- Yan Xue
- Zuoxing Niu
- Kai Cui
- Fachang Yu
- Bo Zhang
- Sheng Li
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Affiliations: Department of Hepatobiliary Surgery, Shandong Cancer Hospital, Jinan 276000, P.R. China, Jinan Child Hospital, Jinan 250011, P.R. China, Key Laboratory for Rare and Uncommon Diseases of Shandong, Jinan 250062, P.R. China, Shandong Academy of Medical Sciences, Jinan 250014, P.R. China
Published online on: February 1, 2013 https://doi.org/10.3892/or.2013.2269
Pages: 1322-1332

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Abstract

In the present study, we investigated the effect of reduced enolase-1 expression in human umbilical vein endothelial cells (HUVECs)/MDA-MB-231 cells on the response to hypoxia and the possible mechanisms involved. Breast cancer cells transfected with enolase-1 siRNA were injected into mice to establish a tumor-bearing mouse model, and the correlation between enolase-1 expression and breast cancer angiogenesis, as well as its effect on the efficacy of radiation therapy were assessed. HUVECs were cultured in vitro, and transfected with enolase-1 siRNA. Following stable passage, 1.0% O2 was used to induce hypoxia. The growth, proliferation, division and angiogenesis of HUVECs were observed using MTT assay, flow cytometry (FCM) and time-lapse video microscopy. The key regulatory molecules were detected using western blot analysis, two-dimensional (2-D) electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The breast cancer cell line, MDA-MB-231, was cultured in vitro, and transfected with enolase-1 siRNA. The cells were injected into nude mice, and radiation therapy was administered. Tumor growth, angiogenesis in tumor tissues and apoptosis were observed, and the expression of the endogenous hypoxia marker, hypoxia inducible factor-1α (HIF-1α), was detected using immunohistochemistry after the mice were sacrificed. A significant reduction in the hypoxia-induced apoptosis of HUVECs was observed in the control group compared with the endothelial cells transfected with enolase-1 siRNA. After the enolase-1 transfected breast cancer cells were injected into nude mice, tumor growth significantly declined, and the tumor volume and weight were reduced. Following treatment with radiation therapy, tumor size significantly decreased in both groups, and the highest reduction was observed in the transfected group. The reduction in enolase-1 expression significantly decreases the response to hypoxia and enhances the sensitivity of the cells to radiation therapy; therefore, enolase-1 may be a drug target for the treatment of breast cancer.

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