We investigated the inhibition of apoptosis and proliferation of poorly differentiated

We investigated the inhibition of apoptosis and proliferation of poorly differentiated AGS gastric cancer cells by epigallocatechin-3-gallate (EGCG), to establish target genes for rules by EGCG. of poorly differentiated AGS gastric cancer cells, and Id1 may be one of the target genes regulated by EGCG in cancer inhibition. … Physique 3. Western blot analysis showed that Id1 protein manifestation was significantly reduced in AGS cells treated with EGCG in a concentration-dependent manner. Id1 protein manifestation was significantly reduced after treatment with 80 g/ml EGCG. -actin … buy 31282-04-9 Detection of Id1 with CCK-8 to observe influence of RNAi on AGS cell proliferation The CCK-8 experiment and cell morphological observation buy 31282-04-9 showed that siRNA-Id1 inhibited proliferation of AGS cells in a concentration-dependent manner ( Fig. 4 , P<0.01). Proliferation of AGS cells was significantly inhibited by 80 nM siRNA-Id1. Physique 4. The CCK-8 experiment showed that Id1 RNAi affected AGS cell proliferation. The inhibition rates for cells treated with siRNA-Id1 at different concentrations and 100 nM siRNA controls were significantly different (P<0.01). siRNA-Id1 inhibited ... Id1 manifestation in AGS cells treated with Id1 RNAi Real-time RT-PCR showed that mRNA manifestation of Id1 was significantly downregulated in AGS cells treated with siRNA-Id1 in a concentration-dependent manner ( Fig. 5 , P<0.01). Western blot analysis further showed that Id1 protein manifestation in AGS cells was consistent with mRNA manifestation ( Fig. 6 ). Physique 5. Real-time RT-PCR showed that Id1 mRNA manifestation in AGS cells treated with siRNA-Id1 was significantly reduced in a concentration-dependent manner. Id1 mRNA manifestation in AGS cells was significantly reduced after treatment with 80 g/ml EGCG. … Physique 6. Western blot analysis showed that Id1 protein manifestation was significantly buy 31282-04-9 reduced in AGS cells treated with siRNA-Id1 CDKN1A in a concentration-dependent manner. -actin was used as a loading control. Apoptosis and cell cycle of AGS gastric cancer cells treated with EGCG and Id1 RNAi Flow cytometry showed that EGCG and siRNA-Id1 induced apoptosis of AGS cells in a concentration-dependent manner ( Fig. 7 ). After treatment with Id1-RNAi, changes in AGS cell proliferation and apoptosis were comparable to those in cells treated with EGCG. AGS cells were arrested at S phase after treatment with EGCG and siRNA-ID1 ( Fig. 8 , P<0.05). Physique 7. Effect of EGCG and Id1 RNAi on AGS gastric cancer cell apoptosis. Flow cytometry showed that EGCG and siRNA-Id1 concentration-dependently induced apoptosis of AGS cells. Apoptosis and necrosis gradually increased in AGS cells treated with different concentrations ... Physique 8. EGCG and Id1 RNAi affected the AGS cell cycle. AGS cells treated with EGCG and siRNA-Id1 were both arrested at S phase (P<0.05). Proliferation and apoptosis of AGS cells treated with Id1-RNAi were comparable to the changes in cells treated with ... Discussion An increasing number of studies have shown that EGCG regulates transduction of signaling molecules related to tumor cell metastasis and migration, and inhibits tumor cell proliferation and induces apoptosis and cell cycle arrest ( 13 , 14 ) . In the present study, CCK-8 experiments and morphological observation showed that EGCG inhibited proliferation of AGS gastric cancer cells in a concentration-dependent manner. Flow cytometry showed that EGCG concentration-dependently induced apoptosis of AGS cells and cell cycle arrest at S phase. We further used gene manifestation microarray analysis for initial screening of differentially expressed genes. There were 54 differentially expressed genes when comparing buy 31282-04-9 EGCG-treated and normal control cells, with 37 upregulated and 17 down-regulated genes. Han et al have exhibited that Id1 manifestation is usually often high in gastric cancer tissues and cell lines and its manifestation level is usually related to the degree of malignancy ( 15 ) . Therefore, high Id1 manifestation is usually directly related to the malignant potential of gastric cancer cells. Tsuchiya et al have also found that gastric cancer cells with high Id1 manifestation have strong metastatic ability ( 16 ) . As far as we know, the role of Id1 in buy 31282-04-9 EGCG-induced.