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1. 潍坊医学院研究生部,山东,潍坊,261000
2. 潍坊医学院附属医院消化内科,山东,潍坊,261000
网络出版:2015-05-08,
纸质出版:2015-05-08
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郭爱,季万胜,姜琪琪等. Δ133p53异构体在5-FU抑制胃癌MKN45细胞系生长实验中的作用[J]. 中国癌症杂志, 2015, 25(1): 25-30.
郭爱, 季万胜. The role of Δ133p53 during 5-FU inhibition experiments on the growth of gastric cancer cell line MKN45[J]. China Oncology, 2015, 25(1): 25-30.
郭爱,季万胜,姜琪琪等. Δ133p53异构体在5-FU抑制胃癌MKN45细胞系生长实验中的作用[J]. 中国癌症杂志, 2015, 25(1): 25-30. DOI: 10.3969/j.issn.1007-3969.2015.01.005.
郭爱, 季万胜. The role of Δ133p53 during 5-FU inhibition experiments on the growth of gastric cancer cell line MKN45[J]. China Oncology, 2015, 25(1): 25-30. DOI: 10.3969/j.issn.1007-3969.2015.01.005.
背景与目的:Δ133p53具有促进肿瘤细胞生长的作用,但具体作用机制尚不明确,本实验是采用5-FU-MKN45胃癌细胞系模型,观察p53异构体Δ133p53表达与p53基因下游MDM2、cyclin G1基因表达的相关性。方法:使用不同浓度5-FU(50 μg/mL,100 μg/mL)作用于人胃癌MKN45细胞系后,MTT法检测细胞抑制率,巢式逆转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR法)检测Δ133p53、MDM2及cyclin G1 mRNA的表达变化。组间差异用单因素方差分析,组内比较用t检验,两变量相关性用Pearson直线相关分析。结果:MTT结果显示,随着5-FU浓度增高以及作用时间的延长,细胞抑制率逐渐增加,4组实验中50 μg/mL 5-FU作用于MKN45细胞24、48和78 h后抑制率平均值分别为41.10%、54.79%和68.48%,差异有统计学意义(F=45.52,P=0.00)。100 μg/mL 5-FU作用于MKN45细胞24、48和78 h后抑制率平均值分别为69.53%、78.21%和86.92%,差异有统计学意义(F=85.58,P=0.00)。50和100 μg/mL 5-FU作用于MKN45细胞24 h后抑制率分别为41.10%和69.53%,差异有统计学意义(F=51.29,P=0.00)。50和100 μg/mL 5-FU作用于MKN45细胞48 h后抑制率分别为54.79%和78.21%,差异有统计学意义(F=51.29,P=0.00)。50和100 μg/mL 5-FU作用于MKN45细胞72 h后抑制率分别为68.48%和86.82%,差异有统计学意义(F=104.91,P=0.00)。RT-PCR结果显示,随着5-FU浓度的增加,胃癌细胞系MKN45细胞中Δ133p53 mRNA、MDM2 mRNA和cyclin G1 mRNA表达量逐渐下降,组间差异具有统计学意义(F值分别为738.532、1 396.607和2 785.560,P=0.00)。相关性分析显示,Δ133p53 mRNA与MDM2 mRNA在胃癌中表达呈正相关(r=0.871,P=0.01),而cyclin G1 mRNA的表达与Δ133p53 mRNA没有明显的相关性(P=0.13)。结论:在5-FU-MKN45模型中,Δ133p53参与的5-FU的抗肿瘤途径与MDM2有关,与cyclin G1无关。
Background and purpose: Δ133p53 can promote tumor cell growth
but the exact mechanism is not clear. This study was aimed to observe the expression and significant of the p53 isoforms Δ133p53 and p53 gene downstream molecules MDM2 and cyclin G1 genes by 5-FU-MKN45 gastric cancer cell line model. Methods: After using different concentrations of 5-FU (50 μg/mL
100 μg/mL) to human gastric cancer cell line MKN45
inhibition rate should be detected by MTT assay
the changes of Δ133p53 mRNA
MDM2 mRNA and cyclin G1 mRNA expressing were detected by reverse transcription-polymerase chain reaction (RT-PCR). Differences between these groups were analyzed by ANOVA
comparisons within groups were analyzed by t-test
bivariate correlation was analyzed by Pearson linear correlation. Results: MTT results showed that with the increased concentration of 5-FU and the extension of time
the cell inhibition rates increased gradually. The inhibition rates of 50 μg/mL 5-FU were 41.10%
54.79% and 68.48%
for culturing 24
48 and 72 hours. There were statistically significant differences between the groups(F=45.52
P=0.00). The inhibition rates of 100 μg/mL 5-FU were 69.53%
78.21% and 86.92%
for culturing 24
48 and 72 hours. There were statistically significant differences between the groups(F=85.58,P=0.00). The inhibition rates of 50 and 100 μg/mL were 41.10% and 69.53%
for culturing 24 hours. There were statistically significant differences between the groups(F=51.29
P=0.00). The inhibition rates of 50 and 100 μg/mL were 54.79% and 78.21%
for culturing 48 hours. There were statistically significant differences between the groups(F=51.29
P=0.00). The inhibition rates of 50 and 100 μg/mL were 68.48% and 86.82%
for culturing 72 hours. There were statistically significant differences between the groups(104.91
P=0.00). RT-PCR results showed that with the increase of the concentration of 5-FU
the Δ133p53 mRNA
MDM2 mRNA and cyclin G1 mRNA expression gradually declined in gastric cancer cell line MKN45 cells
and there were statistically significant differences between the groups(F=738.532
1 396.607
2 785.56,P=0.00). Correlation analysis showed that the expressions of Δ133p53 mRNA and MDM2 mRNA in gastric cancer were positively correlated (r=0.871
P=0.01)
while the expression of cyclin G1 mRNA and p53 mRNA had no obvious relevance (P=0.13). Conclusion: In the 5-FU-MKN45 gastric cancer cell line model
anti-tumor pathway of Δ133p53 isomers is related with MDM2 but was not related with cyclin G1.
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