Effect of broccoli bioactives on prostate cancer cell invasion in vitro

Huang, Ching-Yu

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https://hdl.handle.net/2142/50403 Copy

Description

Title

Effect of broccoli bioactives on prostate cancer cell invasion in vitro

Author(s)

Huang, Ching-Yu

Issue Date

2014-09-16

Director of Research (if dissertation) or Advisor (if thesis)

Erdman, John W.

Department of Study

Food Science & Human Nutrition

Discipline

Food Science & Human Nutrition

Degree Granting Institution

University of Illinois at Urbana-Champaign

Degree Name

M.S.

Degree Level

Thesis

Keyword(s)

• Human prostate cancer cell
• DU145
• invasion
• epithelial-mesenchymal transition
• broccoli bioactives
• sulforaphane
• indole-3-carbinol
• neoglucobrassicin
• quercetin
• 3,3'-indolylmethane

Abstract

Previous studies have shown that broccoli bioactives may have anti-cancer activity. In this work, we hypothesized that broccoli bioactive components inhibit invasion of human prostate cancer cell line, DU145, in an in vitro model system. DU145 cells were plated into Matrigel invasion chambers placed in 24-well plates with complete growth medium as a chemo-attractant. In the chambers, cells were treated with sulforaphane (SF), indole-3-carbinol (I3C), hydrolyzed neoglucobrassicin (hNGB), and quercetin (Q) individually or in selected combinations. After a total of 48 hour treatment exposure, with the treatments being renewed at the 24th hour, the invading cells were fixed, stained, and counted. The results of Matrigel invasion assay indicate that the invasive ability of DU145 cells was significantly inhibited by the physiologically relevant levels of SF (3-5 µM) and I3C (120-200 nM), but not NGB (1-50 µM) or Q (1-5 µM). We further performed the MTT assay to examine whether the reduction of invading cells were attributed to the decreased invasive ability or cell death in the condition of the Matrigel invasion assay. DU145 cells were plated in 96-well plates and followed the treatment and incubation procedure similar to the Matrigel invasion assay, with invasion-suppressive concentrations of SF (3-5 µM) or I3C (160 and 200 nM). At the end of 48 hours, MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) was added to each well, followed by incubation for 4-6 hours. The 96-well plates were analyzed at the absorbance of 540nm. The result of MTT assay shows that invasion-suppressive concentrations of SF and I3C did not decrease cell viability, meaning that SF and I3C inhibited DU145 cells not by causing cell death but by other regulatory pathways that may suppress invasion. The results of this study provide the support for broccoli consumption by prostate cancer patients. Further work will be needed to evaluate whether broccoli bioactives reduce metastasis of prostate cancer to extra-prostatic sites.

Graduation Semester

2014-08

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http://hdl.handle.net/2142/50403

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Copyright 2014 Ching-Yu Huang

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