Lipophilized Epigallocatechin Gallate Derivative Exerts Anti-Proliferation Efficacy through Induction of Cell Cycle Arrest and Apoptosis on DU145 Human Prostate Cancer Cells

Abstract

Epigallocatechin gallate (EGCG) is the predominant tea polyphenol and it exhibits a hydrophilic character. The lipophilized EGCG derivative (LEGCG) was synthesized by enzymatic esterification of EGCG with lauric acid to enhance its bioactivity. The tetralauroyl EGCG was confirmed by high-performance liquid chromatography-tandem mass spectrometry and further identified as 3 , 5 , 3 , 5 -4-O-lauroyl EGCG by 1 H and 13 C nuclear magnetic resonance. The anti-proliferation effect of LEGCG on DU145 human prostate carcinoma cells was evaluated by MTT assay. In addition, the underlying molecular mechanism by which LEGCG exerts anti-proliferation efficacy was elucidated by flow cytometry and immunoblot analysis. Results suggested that LEGCG exhibited a dose-dependent anti-proliferation effect on DU145 cells by G0/G1 phase arrest and induction of apoptosis. LEGCG induced cell cycle arrest via p53/p21 activation, which down-regulated the cyclin D1 and CDK4 expression. In addition, LEGCG induced apoptosis by increasing the Bax/Bcl-2 ratio, the cytochrome c release, and the caspases cleavage on DU145 cells. The results provide theoretical support to prevent prostate cancer with LEGCG.Nutrients 2020, 12, 92 2 of 10 cancer affects [16]. Therefore, we hypothesized that LEGCG has the potential to prevent prostate cancer. In the present study, enzymatic esterification of EGCG was implemented. The high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) and 1 H and 13 C nuclear magnetic resonance (NMR) were used to determine the chemical structures of LEGCG.The inhibitory effect and the underlying mechanism of LEGCG on DU145 cells were investigated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay, flow cytometry, and immunoblot analysis. Materials and Methods MaterialsEGCG, MTT, and propidium iodide (PI) were purchased from Sigma-Aldrich Co. (St. Louis, MO., USA). Annexin V-FITC/PI kit was from Biolegend (San Diego, CA, USA). Cyclin D1, CDK 4, p21, p27, p53, Bax, Bcl-2, cytochrome c, cleaved caspase-9, cleaved caspase-3, and GAPDH antibodies were from Cell Signaling Technology (Danvers, MA, USA). Preparation of Crude EGCG DerivativeLEGCG was prepared according to the literature [17]. Briefly, EGCG and lauric acid were mixed at a mole ratio of 1:1 in ethanol, and then 5% of Lipozyme TLIM (5% w/w) was added. The mixture was heated at 45 • C for 12 h in a screw-capped glass bottle. The reaction was terminated by the removal of the enzyme through filter paper, and the product was concentrated to get crude LEGCG.

EGCG sensitizes chemotherapeutic-induced cytotoxicity by targeting the ERK pathway in multiple cancer cell lines

“…The apoptotic effect was not restrained to only pancreatic cancer cells, since EGCG also strongly induced apoptosis in colon and lung cancer cells, showing that this effect is observed in multiple cancer types. Our findings are consistent with others, showing that EGCG strongly induces apoptosis in various types of tumors [21][22][23][24].…”

Lipophilic Grape Seed Proanthocyanidin Exerts Anti-Cervical Cancer Effects in HeLa Cells and a HeLa-Derived Xenograft Zebrafish Model

“…After treatment with various concentrations of LGSP (50, 100 and 200 µg/mL) for 48 h, cells were harvested and lysed in a RIPA buffer. The total protein concentration was detected using an BCA protein assay kit [ 21 ]. The cell lysates were denatured in a SDS sample buffer and subjected to SDS-PAGE gel.…”

Pharmacological Effects and Potential Clinical Usefulness of Polyphenols in Benign Prostatic Hyperplasia

“…In addition, this study showed that BPH-1 cell migration was significantly inhibited by 10–100 μM EGCG in a wound healing assay [ 95 ]. The anti-proliferative effects of green tea polyphenol were expected according to previous reports on malignancies, including prostate cancer [ 96 , 97 , 98 ]. However, EGCG can also modulate BPH-1 cell migration, although BPH is a benign disease.…”