桑色素对SO-Rb50细胞增殖和转移能力的抑制作用研究

Inhibitory effect of morin on proliferation and metastasis of SO-Rb50 cells

目的研究桑色素对视网膜母细胞瘤(Rb)细胞系SO-Rb50细胞增殖和转移能力的抑制作用及其潜在机制。方法将人Rb细胞系SO-Rb50细胞分为对照组(CG),给予等体积培养液、阿霉素组(AG),给予2.5μg/mL阿霉素、桑色素低剂量组(LCG)、桑色素中剂量组(MCG)和桑色素高剂量组(HCG),每组设3个复孔,分别加入200μM、400μM、600μM浓度的桑色素。采用EDU试剂盒检测细胞增殖;流式细胞仪检测细胞凋亡;Transwell检测细胞侵袭;蛋白免疫印迹试剂盒检测Ki67、Caspase-3、血管内皮生长因子(VEGF)的表达。结果(1)细胞增殖:5组间GEDU阳性细胞数比较,差异有统计学意义(F=57.123,P=0.000)。与CG比较,AG、LCG、MCG、HCG均降低,差异均有统计学意义(P<0.05)。与AG比较,LCG、MCG、HCG均升高,差异均有统计学意义(P<0.05)。与LCG比较,HCG降低,差异有统计学意义(P<0.05)。与MCG比较,HCG降低,差异有统计学意义(P<0.05)。(2)细胞凋亡:5组间细胞凋亡率比较,差异有统计学意义(F=111.168,P=0.000)。与CG比较,AG、LCG、MCG、HCG均升高,差异均有统计学意义(P<0.05)。与AG比较,LCG、MCG、HCG均降低,差异均有统计学意义(P<0.05)。与LCG比较,MCG、HCG均升高,差异均有统计学意义(P<0.05)。与MCG比较,HCG升高,差异有统计学意义(P<0.05)。(3)细胞侵袭:5组间侵袭细胞数比较,差异有统计学意义(F=81.325,P=0.000)。与CG比较,AG、MCG、HCG均升高,差异均有统计学意义(P<0.05)。与AG比较,LCG、MCG、HCG均降低,差异均有统计学意义(P<0.05)。与LCG比较,MCG、HCG均升高,差异均有统计学意义(P<0.05)。与MCG比较,HCG升高,差异有统计学意义。(4)Caspase-3蛋白表达:5组间比较,差异有统计学意义(F=117.778,P=0.000)。与CG比较,AG、LCG、MCG、HCG均升高,差异均有统计学意义(P<0.05)。与AG比较,LCG、MCG均降低,差异均有统计学意义(P<0.05)。与LCG比较,MCG、HCG均升高,差异均有统计学意义(P<0.05)。与MCG比较,HCG升高,差异有统计学意义(P<0.05)。(5)Ki67蛋白表达:5组间比较,差异有统计学意义(F=124.000,P=0.000)。与CG比较,AG、LCG、MCG、HCG蛋白表达均降低,差异均有统计学意义(P<0.05)。与AG比较,LCG、MCG、HCG均升高,差异均有统计学意义(P<0.05)。与LCG比较,HCG降低,差异有统计学意义(P<0.05)。与MCG比较,HCG降低,差异有统计学意义(P<0.05)。(6)VEGF蛋白表达:5组间比较,差异有统计学意义(F=114.747,P=0.000)。与CG比较,AG、LCG、MCG、HCG均降低,差异均有统计学意义(P<0.05)。与AG比较,LCG、MCG、HCG均升高,差异均有统计学意义(P<0.05)。与LCG比较,MCG、HCG均降低,差异均有统计学意义(P<0.05)。与MCG比较,HCG降低,差异有统计学意义(P<0.05)。结论桑色素对Rb细胞系SO-Rb50细胞增殖和转移能力具有抑制作用,并且以浓度依赖的方式促进Caspase-3蛋白表达,抑制Ki6z7、VEGF蛋白表达。

OBJECTIVE To investigate the inhibitory effect of morin on proliferation and metastasis of retinoblastoma(Rb)cell line SO-Rb50 and its potential mechanism.METHODS Human Rb cell line SO-RB50 cells were divided into control group(CG),which were given equal volume culture medium,adriamycin group(AG),which were given 2.5μg/mL adriamycin,morin low-dose group(LCG),morin medium-dose group(MCG)and morin high-dose group(HCG).Each group was administered with 3 multiple holes,and 200μM,400μM and 600μM morin were added respectively.Cell proliferation was detected by EDU kit;Cell apoptosis was detected by flow cytometry;Cell invasion was detected by Transwell;And the expressions of Ki67,Caspase-3 and vascular endothelial growth factor(VEGF)were detected by Western blotting kit.RESULTS(1)Cell proliferation:There was a statistically significant difference in the number of GEDU positive cells among 5 groups(F=57.123,P=0.000).Compared with that in CG,the number of GEDU positive cells in AG,LCG,MCG and HCG decreased,and the differences were all statistically significant(P<0.05).Compared with that in AG,the number of GEDU positive cells in LCG,MCG and HCG increased,and the differences were all statistically significant(P<0.05).Compared with that in LCG,the number of GEDU positive cells in HCG decreased,and the difference was statistically significant(P<0.05).Compared with that in MCG,the number of GEDU positive cells in HCG decreased,and the difference was statistically significant(P<0.05).(2)Cell apoptosis:There was significant difference in the apoptosis rate among the 5 groups(F=111.168,P=0.000).Compared with that in CG,the apoptosis rate in AG,LCG,MCG and HCG increased,and the differences were all statistically significant(P<0.05).Compared with that in AG,the apoptosis rate in LCG,MCG and HCG decreased,and the differences were all statistically significant(P<0.05).Compared with that in the apoptosis rate in LCG,MCG and HCG increased,and the differences were all statistically significant(P<0.05).Compared with that in MCG,the apoptosis rate in HCG increased,and the difference was statistically significant(P<0.05).(3)Cell invasion:There was significant difference in the number of invasive cells among the five groups(F=81.325,P=0.000).Compared with that in CG,the number of invasive cells in AG,MCG and HCG increased,and the differences were all statistically significant(P<0.05).Compared with that in AG,the number of invasive cells in LCG,MCG and HCG decreased,and the differences were all statistically significant(P<0.05).Compared with that in LCG,the number of invasive cells in MCG and HCG increased,and the differences were all statistically significant(P<0.05).Compared with that in MCG,the number of invasive cells in HCG increased,and the difference was statistically significant(P<0.05).(4)Caspase-3 protein expression:The difference between the 5 groups was statistically significant(F=117.778,P=0.000).Compared with that in CG,the Caspase-3 protein expression in AG,LCG,MCG and HCG increased,and the differences were all statistically significant(P<0.05).Compared with that in AG,the Caspase-3 protein expression in LCG,MCG decreased,and the differences were both statistically significant(P<0.05).Compared with that in LCG,the Caspase-3 protein expression in MCG and HCG increased,and the differences were both statistically significant(P<0.05).Compared with that in MCG,the Caspase-3 protein expression in HCG increased,and the difference was statistically significant(P<0.05).(5)Ki67 protein expression:The difference between the 5 groups was statistically significant(F=124.000,P=0.000).Compared with that in CG,the Ki67 protein expression in AG,LCG,MCG and HCG decreased,and the differences were all statistically significant(P<0.05).Compared with that in AG,the Ki67 protein expression in LCG,MCG and HCG increased,and the differences were all statistically significant(P<0.05).Compared with that in LCG,the Ki67 protein expression in HCG decreased,and the difference was statistically significant(P<0.05).Compared with that in MCG,the Ki67 protein expression in HCG decreased,and the difference was statistically significant(P<0.05).(6)VEGF protein expression:The difference between the 5 groups was statistically significant(F=114.747,P=0.000).Compared with that in CG,the VEGF protein expression in AG,LCG,MCG and HCG all decreased with statistical significance(P<0.05).Compared with that in AG,the VEGF protein expression in LCG,MCG and HCG increased,and the differences were statistically significant(P<0.05).Compared with that in LCG,the VEGF protein expression in MCG and HCG both decreased with statistical significance(P<0.05).Compared with that in MCG,the VEGF protein expression in HCG decreased,the difference was statistically significant(P<0.05).CONCLUSIONS Morin inhibited the proliferation and metastasis of Rb cell line SO-RB50,promoted the expression of Caspase-3 protein and inhibited the expression of Ki67 and VEGF protein in a concentration-dependent manner.