青藤碱联合氟尿嘧啶对人胃腺癌SGC7901细胞增殖及凋亡的影响
The effects of sinomenine combined with 5-fluomuracil on the proliferation and apoptosis in human gastric gland carcinoma cell line SGC 7901
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摘要: 目的:探讨青藤碱联合氟尿嘧啶(5-Fu)对人胃癌SGC7901细胞增殖及Bcl-2、Bax和环氧化酶(COX)-2表达的影响。方法:体外培养人胃腺癌SGC7901细胞系,四甲基偶氮唑蓝法测定对照组、5-Fu组(5-Fu 50 mg/L)、青藤碱低浓度组(青藤碱 1.25 mmol/L)、青藤碱中浓度组(青藤碱2.5 mmol/L)、青藤碱高浓度组(青藤碱 5 mmol/L)及联合组(5-Fu 25 mg/L+青藤碱2.5 mmol/L)对SGC7901细胞增殖作用的影响;碘化丙啶染色流式细胞术检测细胞周期变化及凋亡率;免疫细胞化学法测定Bcl-2、Bax和COX-2蛋白表达情况。结果:青藤碱、5-Fu作用SGC7901细胞24、48和72 h后,青藤碱对人胃腺癌SGC7901细胞的增殖具有抑制作用,呈时间、浓度依赖性,与5-Fu联用后协同抑制细胞增殖(P<0.05～P<0.01);5-Fu组、青藤碱各浓度组及联合组的细胞凋亡率均较对照组明显增高(P<0.01),其中联合组均较单药组的凋亡率明显降低(P<0.01)。对照组Bcl-2和COX-2蛋白表达较明显,而青藤碱组、5-Fu组及其联合组染色减弱;与对照组差异均有统计学意义(P<0.05～P<0.01),其中联合组均较单药组阳性率明显降低(P<0.01)。对照组Bax蛋白表达较弱,青藤碱组、5-Fu组及其联合组阳性率相对增加;与对照组比较差异均有统计学意义(P<0.01),且联合组均较单药组阳性率明显升高(P<0.01)。结论:青藤碱联合5-Fu对人胃腺癌SGC7901细胞增殖具有协同抑制作用,能够上调Bax表达,下调Bcl-2、COX-2表达,可能发挥增敏作用。Abstract: Objective:To explore the effects of sinomenine combined with 5-fluomuracil(5-Fu) on the proliferation of human gastric gland carcinoma cell line SGC 7901 and expressions of Bcl-2,Bax and COX-2.Methods:The human gastric gland carcinoma cell line SGC 7901 was cultured in vitro,which was divided into the control group,5-Fu group(50 mg/L),low concentration sinomenine group(1.25 mmol/L),medium concentration sinomenine group(2.5 mmol/L),high concentration sinomenine group(5 mmol/L) and combination group(25 mg/L of 5-Fu and 2.5 mmol/L of sinomenine).The proliferation of SGC7901 cells was detected by MTT,the cell cycle and apoptosis rate were determined by flow cytometry,and the expressions of Bcl-2,Bax and COX-2 were detected by immunohistochemistry.Results:Sinomenine could inhibit the proliferation of SGC7901 cells after 24 h,48 h and 72 h of treatment,which was dose- and time-dependent,and the treatment with sinomenine combined with 5-Fu could strengthen the inhibitory effects(P<0.05 to P<0.01).Compared with the control group,the apoptosis rates in 5-Fu group,sinomenine groups and combination group increased obviously(P<0.01),and the apoptosis rate in combination group was significantly lower than that in single drug group(P<0.05 to P<0.01).The positive expressions of Bcl-2 and COX-2 in the control group,and the sinomenine group,5-Fu group and combination group were strong and weak,respectively,the differences of whose were statistical significance(P<0.05 to P<0.01).The positive rates of Bcl-2 and COX-2 expressions in combination group were lower than those in single drug group(P<0.01 ).The positive expression of Bax in the control group,and the sinomenine group,5-Fu group and combination group were weak and strong,respectively,the differences of whose were statistically significant(P<0.01).The positive rate of Bax expression in combination group was higher than that in single drug group(P<0.01).Conclusions:Sinomenine combined with 5-Fu can enhance the inhibitory effect on SGC7901 cells proliferation,which can further upregulate the expression of Bax,and downregulate the expressions of Bcl-2 and COX-2 protein.
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Key words:
- stomach neoplasms /
- sinomenine /
- 5-fluomuracil /
- cell proliferation /
- apoptosis
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