乌司他丁对顺铂处理的胃癌细胞增殖的影响及作用机制

Effect and Mechanism of Ulinastatin Combined with Cisplatin on Gastric Cancer Cell

目的探讨顺铂联合乌司他丁处理胃癌细胞BGC-823后，观察细胞增殖率和细胞周期相关蛋白的变化。方法通过体外培养胃癌细胞BGC-823。实验分为四组：对照组、顺铂化疗组、乌司他丁组以及顺铂＋乌司他丁组。MTF法检测BGC-823细胞增殖率，观察顺铂及乌司他丁对胃癌细胞系增殖率的影响。WesternBlot法检测胃癌细胞中细胞周期相关蛋白CyclinG1和MEDF2D的表达，RT—PCR法检测细胞中CyclinG1的表达水平。结果细胞增殖率结果表明：与对照组相比，顺铂组、乌司他丁组和顺铂＋乌司他丁组的细胞增殖率明显降低，有显著的统计学意义（P〈0．01）；与顺铂组和乌司他丁组相比，顺铂＋乌司他丁组的细胞增殖率明显降低，有统计学意义（P〈0．01）。WesternBlot结果显示：与对照组相比，顺铂组和乌司他丁组的CyclinG1和MEDF2D蛋白降低（P〈0．05），顺铂＋乌司他丁组的CyclinG1和MEDF2D蛋白亦明显降低，有显著的统计学意义（P〈0．01）；与顺铂组相比，顺铂＋乌司他丁组的CyclinG1和MEDF2D蛋白明显降低，有统计学意义（P〈0．01）。RT—PCR结果表明：与对照组相比，顺铂组和乌司他丁组的CyclinG1的mRNA水平明显降低；与顺铂组和乌司他丁组相比，顺铂＋乌司他丁组的CyclinGI的mRNA水平显著降低（P〈0．01）。结论乌司他丁能降低经顺铂化疗患者的胃癌细胞BGC-823的增殖率，这可能是通过降低细胞周期相关蛋白CyclinG1和MEDF2D的表达所致。

Objective To study the changes of the proliferation and Cyelin protein after the managemeng of the gastric cancer cell BGC - 823 treated by the Cisplatin combined with Ulinastatin. Methods The gastric cancer cell BGC - 823 was cultured in vitro and divided into four groups： the control group, Cisplatin group, Ulinastatin group and Cisplatin ＋ Ulinastatin group. The gastric cancer cell and the proliferation were detected by MTT; The eyelin protein of Cyelin GI and MEDF2D was detected by Western Blot and the level of miRNA Cyelin G1 was detected by RT - PCT. Results MTT showed that compared with control group, the pro- liferation rate was obviously decreased in Cisplatin group, Ulinastatin group and Cisplatin ＋ Ulinastatin group. There was significant difference （ P 〈 0. 01 ）. Compared with Cisplatin group and Ulinastatin group, the proliferation rate was obviously decreased in Cispl- atin ＋ Ulinastatin group. There was statiseally difference （ P 〈 0. 01 ）. Western Blot showed that compared with the control group, the protein of Cyelin G1 and MEDF2D decreased in Cisplatin group and Ulinastatin group （ P 〈 0. 05 ） , and in Cisplatin ＋ Ulinastatin group the protein also obviously decreased （P 〈 0. 01 ）. Compared with Cisplatin group and Ulinastatin group, the levels of Cyclin G1 and MEDF2D were significantly decreased （P 〈 0.01 ）. RT -PCR results showed that compared with the control group, the level of miRNA Cyclin G1 was decreased in Cisplatin group, Ulinastatin group and Cisplatin ＋ Ulinastatin group. Compared with Cisplatin group and Ulinastatin group, the level of miRNA Cyclin G1 was decreased in Cisplatin ＋ Ulinastatin group （P 〈 0. 05）. Conclusion Ulinastatin can decrease the proliferation of gastric cancer cell BGC - 823 and is relate to the decreasing the cyclin protein of Cyclin G1 and MEDF2D.