Antiproliferative and proapoptotic effects of epigallocatechin gallate on human leiomyoma cells

Abstract

Objective-To investigate the effects of epigallocatechin gallate (EGCG), an extract of green tea on cultured human leiomyoma cells (HuLM). Design-Laboratory study.Setting-University hospitals. Patients(s)-Not applicable. Interventions(s)-Not applicable.Main Outcome Measure(s)-HuLM cells were treated with various EGCG concentrations. Cell proliferation was assayed using Hoechst 33258 dye and apoptosis by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Total RNA was isolated and gene expression profiling was performed on 84 key genes related to 18 different signal transduction pathways. The protein levels of PCNA, CDK4, BCL-2 and BAX were examined by Western blot analysis.Result(s)-HuLM cells treated with EGCG showed a dose-and time-dependent inhibition of cell proliferation. TUNEL staining indicated a significant increase in apoptosis in HuLM cells treated with 100 µM of EGCG compared to untreated control (p<0.05). Gene expression profiling indicated that EGCG treatment upregulated representative genes from the TGF-β and stress pathways, while inhibiting the survival pathway and NFκB-dependent inflammatory pathway. Western blotting analysis confirmed that EGCG at ≥50µM significantly decreased the expression of PCNA, CDK4 and BCL-2 as well as increased the expression of the proapoptotic BAX in a dose-dependent manner (p<0.05). Conclusion(s)-EGCG inhibits the proliferation of HuLM cells and induces apoptosis.These results suggest that EGCG may be a potential anti-uterine fibroid agent acting through multiple signal transduction pathways.

Combining the chemotherapeutic effects of epigallocatechin 3-gallate with siRNA-mediated p53 knock-down results in synergic pro-apoptotic effects

“…2 Although earlier studies have presented a correlative role of p53siRNA+EGCG in the reduction of cell proliferation, 4,8,[13][14][15] the current study has unequivocally established the higher efficiency of p53siRNA in the presence of EGCG in cell proliferation reduction. This effect is demonstrated by the MTT data and is confirmed at the molecular level using a PCR-based expression array.…”

“…17 The analysis of mRNA gene expression provides evidence of a distinct expression profile of genes involved in apoptosis. 13 EGCG in combination with p53siRNA has synergic pro-apoptotic effects, which are greater than each agent taken individually and induce an immediate expression of anti-apoptotic genes (mRNA), as shown in Figure S1. In contrast, the expressions of BAD, BAX, BCL2L1, BCL2L2, MCL1, and NOL3 were reduced.…”

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“…По-лученные данные согласуются с моделью, в которой эндотелиальный ингибитор активатора плазминогена функционирует как реостат для регулирования взаи-мосвязанных событий реструктуризации матрицы и клеточной миграции [47]. Таким образом, урокиназа, представляет собой многофункциональный белок, который, помимо регуляции фибринолиза, осущест-вляет активацию факторов роста, модуляцию про-дукции цитокинов, фенотипическую трансформацию клеток, экспрессию белков и активацию протеолити-ческих каскадов [48].…”

“…Доксорубицин является основой большинства терапевтических протоколов при лече-нии мелкоклеточного рака легкого, рака молочной железы [45], гемобластозов, мягкотканых опухолей, [46]. Даунорубицин применяется в основном в тера-пии лейкозов у детей и взрослых [46,47,48].…”

“…Третий эксперимент был проведен нами с примене-нием оптической когерентной томографии, где было также показано, что введение золотых наностержней (70 нм) в подкорковую область приводит к быстрому (через 20 мин) их накоплению в глубоком шейном лимфоузле со стороны области введения [48,49].…”