de la Cruz, Joseph Flores
(Department of Basic Veterinary Sciences, College of Veterinary Medicine, University of the Philippines Los Banos)
,
Vergara, Emil Joseph Sanvictores
(Department of Animal Life and Environmental Science, Hankyong National University)
,
Cho, Yura
(Department of Animal Life and Environmental Science, Hankyong National University)
,
Hong, Hee Ok
(Department of Medical Science, School of Medicine, Konkuk University)
,
Oyungerel, Baatartsogt
(Department of Biotechnology and Breeding, School of Animal Science and Biotechnology, Mongolian University of Life Sciences)
,
Hwang, Seong Gu
(Department of Animal Life and Environmental Science, Hankyong National University)
Glehnia littoralis (GL) is widely used as an oriental medicine for cough, fever, stroke and other disease conditions. However, the anti-cancer properties of GL on MCF-7 human breast cancer cells have not been investigated. In order to elucidate anti-cancer properties and underlying cell death mechan...
Glehnia littoralis (GL) is widely used as an oriental medicine for cough, fever, stroke and other disease conditions. However, the anti-cancer properties of GL on MCF-7 human breast cancer cells have not been investigated. In order to elucidate anti-cancer properties and underlying cell death mechanisms, MCF-7cells ($5{\times}10^4/well$) were treated with Glehnia littoralis root extract at 0-400 ug/ml. A hot water extract of GL root inhibited the proliferation of MCF-7 cells in a dose-dependent manner. Analysis of the cell cycle after treatment of MCF-7 cells with increasing concentrations of GL root extract for 24 hours showed significant cell cycle arrest in the G1 phase. RT-PCR and Western blot analysis both revealed that GL root extract significantly increased the expression of p21 and p27 with an accompanying decrease in both CDK4 and cyclin D1. Our reuslts indicated that GL root extract arrested the proliferation of MCF-7 cells in G1 phase through inhibition of CDK4 and cyclin D1 via increased induction of p21 and p27. In summary, the current study showed that GL could serve as a potential source of chemotherapeutic or chemopreventative agents against human breast cancer.
Glehnia littoralis (GL) is widely used as an oriental medicine for cough, fever, stroke and other disease conditions. However, the anti-cancer properties of GL on MCF-7 human breast cancer cells have not been investigated. In order to elucidate anti-cancer properties and underlying cell death mechanisms, MCF-7cells ($5{\times}10^4/well$) were treated with Glehnia littoralis root extract at 0-400 ug/ml. A hot water extract of GL root inhibited the proliferation of MCF-7 cells in a dose-dependent manner. Analysis of the cell cycle after treatment of MCF-7 cells with increasing concentrations of GL root extract for 24 hours showed significant cell cycle arrest in the G1 phase. RT-PCR and Western blot analysis both revealed that GL root extract significantly increased the expression of p21 and p27 with an accompanying decrease in both CDK4 and cyclin D1. Our reuslts indicated that GL root extract arrested the proliferation of MCF-7 cells in G1 phase through inhibition of CDK4 and cyclin D1 via increased induction of p21 and p27. In summary, the current study showed that GL could serve as a potential source of chemotherapeutic or chemopreventative agents against human breast cancer.
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문제 정의
This study aims to determine the cytotoxic effect of the hot water extract of Glehnia littoralis roots on MCF-7 human breast cancer cell line. In addition, the mechanism of action of the extract related to its cytotoxicity was elucidated through the detection of cell cycle-related genes and proteins.
제안 방법
Afterwards, the wells were treated with different concentrations of GL extract (0, 50 100, 200 and 400 µg/mL) for 24 hours.
대상 데이터
The cells were then treated with RNAse A (50 µg/mL) and propidium iodide (50 µg/mL) prior to analysis using FACSCalibur Flow Cytometer (Becton Disckson, USA). 10000 events were evaluated for all samples. The percentage of cells in G0/G1, S and G2/M phases were calculated using BD Cellquest Pro software.
1 µg of RNA was used to obtain the complementary DNA (cDNA) using the protocol provided by M-MuLV reverse transcriptase (Fermentas, Lithuania). Specific primers (Table 1) were used to amplify the genes p21, p27, CDK4 and Cyclin D1. PCR products were then separated by electrophoresis using 2% agarose stained with ethidium bromide and UV transillumination was done afterwards.
데이터처리
All experiments were done in triplicates and data were expressed as means ±SD. The difference between control and GL treated cells were evaluated using one-way ANOVA followed by Duncan Multiple Range Test. P values less than 0.
이론/모형
The viability of MCF-7 breast cancer cells treated with various concentrations of GL extract was measured using CCK-8 assay. As shown in Figure 1, GL extract was able to cause cytotoxicity towards the cells in a dose-dependent manner but not in a time dependent manner (data not shown).
성능/효과
Results obtained from this study demonstrate the cytostatic effect of GL aqueous extract on MCF-7 breast cancer cells through the decreased expression of cell cycle-related genes and proteins. These findings show that Glehnia littoralis roots can be a potential source of a chemotherapeutic or chemopreventative agent against breast cancer.
5% for the control and 6% for the 400µg/mL of treatment. The FACS data shown in this study demonstrated that GL extract inhibited cell proliferation of MCF-7 cells during the G0/G1 phase of the cell cycle.
The results of the cell viability assay have shown a dose dependent decrease in the cell line used in the study with as much as 68.53%, 55.15%, 47.38% and 39.57% reduction in the cells at 50, 100, 200 and 400 μg/mL concentration of the GL extract, respectively, after 24 hours of treatment.
참고문헌 (16)
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