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干扰Sema4D基因表达对胰腺癌细胞生物学特征的影响
Effect of interference of Sema4D expression on cell biological characteristics in pancreatic cancer
文章发布日期:2018年01月05日  来源:  作者:侯丽艳,贾如江,任利兵,等  点击次数:326次  下载次数:60次

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【摘要】:目的探讨干扰Sema4D基因的表达对胰腺癌细胞生物学的影响。方法设计合成Sema4D-siRNA,转染到人胰腺癌细胞系中,将实验分为3组,转染siRNA组、阴性对照组(即转染非特异性对照组)﹑空白对照组(未经任何处理的胰腺癌细胞)。转染48 h后利用RT-qPCR检测转染前后Sema4D mRNA表达的变化,转染72 h后利用Western-Blot的方法检测转染前后Sema4D蛋白表达的变化;采用四甲基偶氮唑蓝显色法观察转染后细胞生长变化;Transwell小室侵袭试验检测转染后肿瘤细胞侵袭性的改变;采用流式细胞技术检测肿瘤细胞凋亡的变化。计量资料多组间比较采用单因素方差分析比较,进一步两两比较采用LSD-t检验法。结果转染Sema4D-siRNA 48 h后,转染siRNA组Sema4D mRNA的表达明显低于阴性对照组和空白对照组,3组比较差异有统计学意义(F=421.990,P<0.001)。转染Sema4D-siRNA 72 h后,转染siRNA组Sema4D蛋白明显低于阴性对照组和空白对照组,3组间比较差异有统计学意义(F=27.074,P=0.002 3)。转染siRNA组48、72、96 h的细胞生长速度明显低于阴性对照组和空白对照组,3组间比较差异均有统计学意义(F值分别为15.314、62.255、223.493,P值分别为0.004、<0.001、<0.001)。转染siRNA组穿膜细胞个数明显低于阴性对照组和空白对照组,3组间比较差异有统计学意义(42.0±5.9 vs 60.0±6.1 vs 61.0±4.6,F=37.21,P=0.000 4]。转染siRNA组穿膜细胞凋亡率明显低于阴性对照组和空白对照组,3组间比较差异有统计学意义[(16.57±0.31)% vs (9.50±0.45)% vs (9.90±0.61)%,F=26.75,P=0.007 4]。结论siRNA能够下调Sema4D基因在胰腺细胞中的表达,可以抑制胰腺癌细胞增殖,使胰腺癌细胞侵袭能力明显下降,凋亡率下降。
【Abstract】:ObjectiveTo investigate the effect of interference of Sema4D expression on cell biological behavior in pancreatic cancer. MethodsSema4D-siRNA was designed, synthesized, and transfected into a human pancreatic cancer cell line. The cells were divided into siRNA transfection group, negative control group (nonspecific transfection group), and blank control group (untreated pancreatic cancer cells). RT-qPCR was used to measure the change in the mRNA expression of Sema4D at 48 hours after transfection, and Western blot was used to measure the change in the protein expression of Sema4D at 72 hours after transfection. MTT assay was used to observe the change in cell growth after transfection, Transwell chamber invasion assay was used to observe the change in tumor cell invasion after transfection, and flow cytometry was used to observe the change in the apoptosis of tumor cells. An analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between any two groups. ResultsAt 48 hours after Sema4D-siRNA transfection, the siRNA transfection group had significantly lower mRNA expression of Sema4D than the negative control group and the blank control group (F=421.990, P<0.001). At 72 hours after Sema4D-siRNA transfection, the siRNA transfection group had significantly lower protein expression of Sema4D than the negative control group and the blank control group (F=27.074, P=0.002 3). At 48, 72, and 96 hours after transfection, the siRNA transfection group had a significantly lower cell growth rate than the negative control group and the blank control group (F=15.314, 62.255, and 223.493, P=0.004, <0.001, and <0.001). The siRNA transfection group had a significantly lower number of cells which crossed the membrane than the negative control group and the blank control group (42.0±5.9 vs 60.0±6.1 vs 61.0±4.6,F=37.21, P=0.000 4). The siRNA transfection group had a significantly lower apoptosis rate than the negative control group and the blank control group [(16.57±0.31)% vs (9.50±0.45)% vs (9.90±0.61)%,F=2675,P=0.007 4]. ConclusionSema4D-siRNA can downregulate the expression of Sema4D in pancreatic cells and can inhibit the proliferation of pancreatic cancer cells and reduce their invasion ability and apoptosis.
【关键字】:胰腺肿瘤; 细胞增殖; 细胞凋亡; 肿瘤侵润; RNA干扰
【Key words】:pancreatic neoplasms; cell proliferation; apoptosis; neoplasm invasiveness; RNA interference
【引证本文】:HOU LY, JIA RJ, REN LB, et al. Effect of interference of Sema4D expression on cell biological characteristics in pancreatic cancer[J]. J Clin Hepatol, 2018, 34(2): 350-353. (in Chinese)
侯丽艳,贾如江,任利兵, 等. 干扰Sema4D基因表达对胰腺癌细胞生物学特征的影响[J]. 临床肝胆病杂志, 2018, 34(2): 350-353.

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