ABSTRACT
Objective: To investigate the effects of the combination of Celastrus orbiculatus extracts and miR-302 on proliferation, invasion and migration of human esophageal cancer cells and the regulation of PI3K/Akt signaling pathway. Methods Quantitative RT-PCR was used to detect the expression of miR-302 in human normal esophageal epithelial cells Het-1A and different esophageal cancer cell lines. The miR-302 mimic and negative control mimic control were transfected into human esophageal cancer TE-1 cells, and qPCR was used to detect the expression of miR-302 in TE-1 cells after plasmid transfection. TE-1 cells were treated with C. orbiculatus extracts alone and combination treatment. The proliferation of TE-1 cells was detected by CCK-8 assay. The invasion and migration of TE-1 cells were detected by Transwell assay. Western blot analysis of the expression of related protein in the PI3K/Akt signaling pathway was carried out. Results: The expression of miR-302 in esophageal cancer cell lines was significantly lower than that in esophageal epithelial cells (P < 0.05). Transfection of miR-302 mimic could effectively increase the expression of miR-302 in TE-1 cells (P < 0.05). The use of C. orbiculatus extracts alone or overexpression of miR-302 inhibited proliferation, invasion and migration of esophageal cancer TE-1 cells (P < 0.05), down-regulated PI3K and p-Akt protein expression; Combination treatment had more significant effect on inhibiting proliferation, invasion and migration of TE-1 cells and down-regulating protein expression of PI3K and p-Akt (P < 0.05). Conclusion: Celastrus orbiculatus extracts combined with miR-302 can synergistically inhibit the proliferation, invasion, and migration of esophageal cancer TE-1 cells, and its mechanism may be related to the inhibition of PI3K/Akt signaling pathway activation.
ABSTRACT
Objective Solamargine (SM), with its anti-inflammatory and anti-tumor effects, inhibits the proliferation and promotes the apoptosis of various tumor cells. This study was to investigate the effects of SM on the proliferation and apoptosis of human esophageal cancer KYSE150 cells and its action mechanism. Methods We treated KYSE150 cells with SM at the concentrations of 0 (the blank control group), 2, 4, 6 and 8 μmol/L for 24 hours. Then, we observed the morphological changes of the cells under the inverted microscope, detected their proliferation and apoptosis by MTT assay and flow cytometry respectively, and determined the expressions of the classical NF-κB signaling pathway-related proteins NF-κB, p-NF-κB, IKKα, IKKβ, IkBα and p-IkBα) and apoptosis-related proteins Bax, caspase-3, cleaved caspase-3 and Bcl-2 in different groups of the cells by Western blot. Results Compared with the blank control, the inhibition rate of the proliferation of the KYSE150 cells in the 2, 4, 6 and 8 μmol/L SM groups was increased significantly in a concentration-dependent manner (0 vs [15.03 ± 0.15]%, [47.94 ± 1.74]%, [68.72 ± 0.47]% and [77.51 ± 1.70]%, P<0.05), and so was the apoptosis rate ([8.17 ± 0.51]% vs [14.50 ± 0.73]%, [18.57 ± 2.08]%, [65.10 ± 10.88]% and [81.55 ± 5.48]%, P<0.05). The expression of the apoptosis-related protein Bax in the SM treated cells was up-regulated, those of Bcl-2, IKKα, IKKβ and p-IkBα down-regulated, and the activity of caspase-3 and cleaved caspase-3 promoted, all in a concentration-dependent manner, with statistically significant differences between the blank control and the 4, 6 and 8 μmol/L SM groups (P<0.05). Statistically significant differences were also found in the expressions of NF-κB, p-NF-κB and IkBα between the blank control and the 6 and 8 μmol/L SM groups (P<0.05). Conclusion Solamargine can significantly inhibit the proliferation and promote the apoptosis of KYSE150 cells, probably by suppressing the classical NF-κB signaling pathway.
ABSTRACT
Objective To observe the effect of Bufalin on controlling ECA109 cells , and to explore its potential anti-tumor mechanism. Method The effect of Bufalin on the proliferation of human esophageal cancer ECA109 cells was evaluated by CCK-8 assay and it effects on apoptosis of esophageal cancer ECA109 cells were determined by flow cytometry (FCM). Protein expressions of NF-κBp65, ERK, PERK1/2 and Caspase-3, Cleaved Caspase-3, PARP, Cleaved PARP in esophageal cancer ECA109 cell were observed through Western blot. Results The proliferation of human esophageal cancer ECA109 cells was significantly inhibited in bufalin group in a time- and concentration-dependent manner. Bufalin can induce the apoptosis in human esophageal ECA109 cells. Results of Western blot showed the protein expressions of apoptosis-related protein Cleaved Caspas-3 and Cleaved PARP in esophageal cancer ECA109 cells could be markedly up-regulated by Bufalin , but p-ERK1/2, NF-κBp65 in esophageal cancer ECA109 cells could be markedly down-regulated by Bufalin. Conclusion Bufalin can potently inhibit the growth of esophageal cancer ECA109 cells and the potential anti-tumor mechanism might be involved in inhibiting ERK/ NF-κB signal pathway.
ABSTRACT
Objective To investigate the effect of SKP2 expression on radiation induced bystander effect (RIBE) of esophageal cancer cells.Methods The esophageal cancer cell lines with different SKP2 levels were applied for the study and the SKP2 expression was identified by Western blot.Micronuclei (MN) assay and DNA foci assay were used to evaluate the effect of SKP2 on RIBE.The cells were transfected with SKP2 gene or SKP2 siRNA to further verify the effect of SKP2 on RIBE.Results MN assay showed that the bystander effect induced by the cells with a high level of SKP2 was lower than that induced by the cells with a lower level of SKP2 (t =8.06,P ＜ 0.01).These results were further confirmed by the gene transfection experiments.When the expression of SKP2 was increased,RIBE was decreased (t=11.12,10.16,P ＜ 0.01).Contrarily,when the expression of SKP2 was reduced,RIBE was increased (t =8.39,8.83,P ＜ 0.01).γ-H2AX foci formation assay disclosed that when SKP2 expression in the irradiated cells increased,the repair ability of DNA damage in the bystander cells was higher than the control (t =6.85,7.10,P ＜ 0.01).With the expression of SKP2 decreased,the repair ability of DNA damage was lower than the control (t =7.66,8.47,P ＜ 0.01).Conclusions Over-expression of SKP2 inhibits RIBE of esophageal cancer cells,at least partly through regulating DNA damage repair ability.