AKAP95、cyclinD1、cyclinE1以及Cx43在 结直肠癌模型小鼠病情发展中的协同调控作用研究

Synergistic regulation of AKAP95,cyclinD1,cyclinE1 and Cx43 in the development of colorectal cancer model mice

目的探讨与研究AKAP95、cyclinD1、cyclinE1、Cx43在结直肠癌模型小鼠病情发展中的协同调控作用。方法无特定病原体级雌性BALB/c裸小鼠32只,将所有小鼠按照随机数字表法分为2组-模型组(n=16)与对照组(n=16)。模型组采用葡聚糖硫酸钠诱导法建立结直肠癌模型,对照组只注射同剂量的0.9%氯化钠溶液。观察小鼠的一般状态,造模后第6周与第12周测量并计算小鼠肿瘤体积和肿瘤占结肠百分比;造模后第12周,采用流式细胞术检测CD3^(+)、CD4^(+)T淋巴细胞比例,蛋白质印迹法检测AKAP95、cyclinD1、cyclinE1以及Cx43蛋白的表达情况。结果造模后第6周与第12周,模型组小鼠的疾病活动指数[(7.22±0.83)、(9.88±0.17)分]均低于对照组[(1.09±0.13)、(1.11±0.15)分],差异均有统计学意义(P<0.05);造模后第6周和第12周,模型组肿瘤体积[(1156.98±123.75)mm 3和(1641.87±154.06)mm 3]均高于对照组(均为0),2组相比差异有统计学意义(P<0.05)。造模后第12周,模型组小鼠的结肠长度[(5.38±0.84)cm]少于对照组[(7.94±0.46)cm],差异有统计学意义(P<0.05),肿瘤占结肠百分比为(25.62±2.48)%。造模后第12周,模型组小鼠的全血CD3^(+)、CD4^(+)T淋巴细胞比例[(33.61±1.11)%、(34.98±1.47)%]低于对照组[(41.56±1.84)%、(38.77±2.11)%],结肠组织AKAP95、cyclinD1、cyclinE1、Cx43蛋白相对表达水平(4.52±0.42、4.29±0.41、3.76±0.28、4.11±0.38)均高于对照组(1.34±0.18、1.22±0.18、1.09±0.23、1.47±0.14),差异均有统计学意义(P<0.05)。Pearson分析显示模型组小鼠结肠组织AKAP95、cyclinD1、cyclinE1、Cx43蛋白相对表达水平与疾病活动指数、结肠长度、CD3^(+)、CD4^(+)T淋巴细胞比例均存在相关性(P<0.05)。结论AKAP95、cyclinD1、cyclinE1、Cx43在结直肠癌模型小鼠中呈现高表达,与小鼠的疾病活动度、病理状况与免疫功能都有一定的相关性。

Objective To explore and study the synergistic regulation of AKAP95,cyclinD1,cyclinE1,and Cx43 in the development of colorectal cancer model mice.Methods specific pathogen free female BALB/c nude 32 mice were divided into two groups-model group(n=16)and control group(n=16)according to the random number table method.In the model group,the colorectal cancer model was established by the dextran sodium sulfate induction method,and the control group was injected with the same dose of normal saline.The general state of the mouse was observed,the tumor volume and the percentage of the recorded tumor in the colon are measured and calculated at the 6th and 12th week of modeling,and the flow cytometry was used to detect the ratio of CD3^(+)and CD4^(+)T lymphocytes at the 12th week of modeling,Western blotting was used to detect the expression of AKAP95,cyclinD1,cyclinE1 and Cx43 proteins.Results The disease activity index of the model group[(7.22±0.83),(9.88±0.17)points]were lower than those of the control group[(1.09±0.13),(1.11±0.15)]at the 6th and 12th week of modeling,the differences were statistically significant(P<0.05).The tumor volume of the model group[(1156.98±123.75)mm 3 and(1641.87±154.06)mm 3]was higher than that of the control group(both 0)at the 6th and 12th week after modeling,the difference between the two groups was statistically significant(P<0.05).The colon length of the model group[(5.38±0.84)cm]was shorter than that of the control group[(7.94±0.46)cm]at the 12th week after modeling,the difference was statistically significant(P<0.05),and the percentage of tumor in the colon was(25.62±2.48)%.The proportion of CD3^(+)and CD4^(+)T lymphocytes in the whole blood of the model group[(33.61±1.11)%,(34.98±1.47)%]was lower than that of the control group[(41.56±1.84)%,(38.77±2.11)%]at the 12th week after modeling,the relative expression levels of AKAP95,cyclinD1,cyclinE1,and Cx43 proteins in colon tissue(4.52±0.42,4.29±0.41,3.76±0.28,4.11±0.38)were higher than those of the control group(1.34±0.18,1.22±0.18),1.09±0.23,1.47±0.14),the differences were all statistically significant(P<0.05).Pearson analysis showed that the relative expression levels of AKAP95,cyclinD1,cyclinE1,and Cx43 protein in the colon tissue of the model group were correlated with disease activity index,colon length,CD3^(+),CD4^(+)T lymphocyte ratio(P<0.05).Conclusion AKAP95,cyclinD1,cyclinE1,and Cx43 are highly expressed in colorectal cancer model mice,which are related to the disease activity,pathological conditions and immune function of the mice.