RESUMEN
Objective:To study the effect of extract of livistona chinensis on the proliferation and apoptosis of bladder cancer cells and the related mechanism.Methods:T24 cells were cultured in medium with the final concentration of 0, 25, 50 and 100 mg/L livistona chinensis extract, respectively. And then they were divided into control group and low, medium and high dose groups. The cell survival rate was detected by cell counting kit 8 (CCK-8). Colony formation assay was used to detect the number of cell clones. Apoptosis was detected by flow cytometry. Western blot was used to detect the expression of related proteins. The Syk overexpression vector plasmid and its negative control were transfected into T24 cells. After transfection, the cells were treated with 100 mg/L livistona chinensis. The cell survival rate, colony formation number and apoptosis rate were detected by the above method. The bladder cancer model nude mice were treated with different concentrations of livistona chinensis extract. Under the microscope, the expression of protein was detected by immunohistochemical staining of bladder tissue.Results:Compared with the control group, the survival rate of T24 cells in the low, medium and high dose livistona chinensis extract groups were significantly decreased [(88.50±3.65)%, (70.58±2.47)%, (48.90±2.37)% vs. (98. 25±4.26)%], and the number of clone formation decreased significantly [(101. 33±3.40), (84.00±2.94), (60.00±2.16) vs. (121.33±4.64) ], and the apoptosis rate was significantly increased [(11.45± 0.59)%, (17.71±0.64)%, (21.33±0.83)% vs. (7. 86±0.43)%]. The expression level of Ki-67 protein was significantly decreased, while the expression levels of Caspase3 and Syk protein were significantly increased in a concentration dependent manner ( P < 0.05). The cell survival rate of pcDNA3.1-Syk group was significantly lower than that of pcDNA3.1 group [(63.87±2.53)% vs. (98. 45±3.54)%], the number of clone formation decreased significantly [(74. 33±2.87) vs. (121.33±3.68)], and the apoptosis rate was significantly increased [(18.39±0.63)% vs. (7.89± 0.45)%] (all P<0.05). The cell survival rate in the high-dose group of livistona chinensis+ pcDNA3.1-Syk was significantly lower than that in the high-dose group of livistona chinenisi+ pcDNA3.1 group [ (29.80±1.63)% vs.(49.33±2.76)% ], the number of clone formation decreased significantly [(33.00±2.94) vs. (59.67±3.30) ], and the apoptosis rate was significantly increased [(26.93±0.68)% vs. (21.25±0.78)% ]( P<0.05). The experimental results of nude mice of bladder cancer model showed that the tumor volume of transplanted bladder cancer nude mice in the control group and the low, medium, and high dose livistona chinensis extract groups were (1 209.75±64.37), (1 006.31±40.49), (530.58±42.87), (267.58±16.73)mm 3, respectively, the weight of the transplanted tumor were (0.36±0.08), (0.30±0.04), (0.26±0.03), (0.18±0.06)g, and the differences between the two groups were statistically significant ( P <0.05). Immunohistochemical staining results showed that the expression of Sky and Caspase3 was increased and the expression of Ki-67 was decreased in the middle and high dose groups compared with that in the control group. Conclusion:Extract of livistona chinensis can inhibit the proliferation and promote apoptosis of bladder cancer cells by up regulating Syk expression.
RESUMEN
The present study aimed to detect the impact of the ethanol extract of the Livistona chinensis seed (EELC) on angiogenesis in human umbilical vein endothelial cells (HUVECs). A chorioallantoic membrane (CAM) assay was used to detect the anti-angiogenic activity of EELC in vivo. In vitro, the effect of EELC on the proliferation, migration and angiogenesis of HUVECs was determined by an MTT assay, a wound healing assay and a tube formation assay, respectively. The vascular endothelial growth factor (VEGF)-A and VEGF receptor (VEGFR)-2 protein and mRNA level were measured with ELISA and reverse transcription-semi-quantitative polymerase chain reaction. It was observed that EELC significantly decreased the formation of new vessels in the CAM assay. EELC inhibited the proliferation and migration of HUVECs. The extent of tube formation by HUVECs was also reduced by EELC. In addition, EELC treatment reduced the level of VEGF-A and VEGFR-2 mRNA and protein. The results suggest that EELC inhibits tumor angiogenesis through inhibiting the proliferation and migration of HUVECs, and by downregulating VEGF and VEGFR.
