尾型同源盒基因2过表达对人胃癌耐顺铂细胞SGC7901/DDP多药耐药性的影响

Effects of overexpression of homeobox gene caudalrelated homeodomain transcription 2 on the multidrug resistance of human gastric cancer cells

    摘要
  • 摘要: 目的：探讨尾型同源盒基因2(Cdx2)过表达对人胃癌耐顺铂细胞SGC7901/DDP的多药耐药性的影响。
    方法：将SGC7901/DDP细胞分为实验组［Cdx2过表达重组慢病毒载体(pCMA Cdx2 HA)处理］、空载体组［慢病毒载体(pCMA HA)处理］、对照组(不做处理)3组。MTT法检测各组SGC7901/DDP细胞对化疗药物的敏感性；流式细胞仪检测各组SGC7901/DDP细胞阿霉素的泵出蓄积量、细胞周期分布及凋亡情况；Western blot技术进一步检测Cdx2蛋白和多药耐药基因1(MDR1)的蛋白表达水平。组间比较采用方差分析。
    结果：SGC7901/DDP细胞感染慢病毒载体72 h后，转染效率达90%以上。实验组Cdx2蛋白的相对表达量为0.37±0.06，较空载体组的0.12±0.02和对照组的0.11±0.03明显升高，3组比较，差异有统计学意义(F=82.37，P<0.05)。实验组SGC7901/DDP细胞对阿霉素、5 氟尿嘧啶和顺铂的半数凋亡浓度分别为(1.10±0.08)mg/L、(4.81±0.12)mg/L、(3.81±0.15)mg/L，均高于空载体组的(0.59±0.05)mg/L、(3.71±0.14)mg/L、(2.20±0.15)mg/L和对照组的(0.63±0.04)mg/L、(3.92± 0.15)mg/L、(2.92±0.11)mg/L，3组比较，差异有统计学意义(F=158.75，98.06，188.78，P<0.05)。实验组阿霉素的泵出率为18.20%±0.12%，高于空载体组的7.22%±0.09%和对照组的8.79%±0.11%，3组比较，差异有统计学意义(F=146.31，P<0.05)。实验组G1期细胞的比例为52.2%±4.4%，明显低于空载体组的62.0%±5.0%和对照组的67.8%±3.3%；实验组S期的细胞比例为40.4%±1.5%，明显高于空载体组的25.6%±2.0%和对照组的21.8%±1.3%，3组细胞G1期和S期比较，差异均有统计学意义 (F=17.08，236.83，P<0.05)。实验组细胞凋亡率为7.6%±1.3%，明显低于空载体组的11.1%±1.1%和对照组的10.8%±1.0%，3组比较，差异有统计学意义(F=16.29，P<0.05)。实验组中MDR1蛋白的表达量为1.85±0.18，高于空载体组的1.12±0.08和对照组的1.02±0.09，3组比较，差异有统计学意义(F=76.22，P<0.05)。
    结论：Cdx2基因的过表达可降低胃癌耐顺铂细胞SGC7901/DDP对化疗药物的敏感性，降低化疗药物在胃癌细胞内的蓄积浓度，增强胃癌多药耐药细胞的耐药性。

     

    Abstract: Objective:To investigate the effects of over expression of caudal related homeodomain transcription 2 (Cdx2) on multi drug resistance of the human gastric cell line SGC7901/DDP.
    Methods:The human gastric cell line SGC7901/DDP was divided into the experimental group (cells were infected with Cdx2 recombinated lentiviral vector), the empty vector group (cells were infected with lentiviral vector) and the control group, respectively. The sensitivity of SGC7901/DDP to the chemotherapy agents was detected by the MTT method. The pump out accumulation rates of adriamycin, cell cycle and apoptosis were detected by flow cytometry. The protein expressions of Cdx2 and multi drug resistance gene 1 (MDR1) were analyzed by Western blot. All data were analyzed using the analysis of variance.
    〖WTHZ〗R0ResultsThe transfection efficiency was above 90% after transfecting the SGC7901/DDP cells by lentiviral vector for 72 hours. The relative expression of Cdx2 protein in the experimental group was 0.37±0.06, which was significantly higher than 0.12±0.02 of the empty vector group and 0.11 ±0.03 of the control group, with no significant difference between the 3 groups (F=82.37, P<0.05). The IC 50 of SGC7901/DDP to adriamycin, 5 fluorouracil and cisplatin in the experimental group were (1.10± 0.08)mg/L, (4.81±0.12)mg/L and (3.81±0.15)mg/L, which were significantly higher than (0.59±R00.05)mg/L, (3.71±0.14)mg/L and (2.20±0.15)mg/L of the empty vector group and (0.63±0.04)mg/L, (3.92± 0.15)mg/L and (2.92±0.11)mg/L of the control group (F=158.75, 98.06, 188.78, P<0.05). The pump out accumulation rate of adriamycin in the experimental group was 18.20% ±1.0% of the control group (F=16.29, P< 0.05). The relative protein expression of MDR1 in the experimental group was 1.85±0.18, which was significantly higher than 1.12±0.08 in the empty vector group and 1.02±0.09 of the control group (F=76.22, P<0.05).
     Conclusion Over expression of Cdx2 gene could decrease the sensitivity of gastric cell line SGC7901/DDP to chemotherapy agents as well as the intracellular accumulation of the agents, and it can also enhance the multi drug resistance of the gastric cell line SGC7901/DDP.

     

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