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；Westernblot技术进一步检测Cdx2蛋白和多药耐药基因1（MDRl）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72h后，转染效率达90％以上。实验组Cdx2蛋白的相对表达量为0．374-0．06，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、（2．92-4-0．11）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％4-0．11％，3组比较，差异有统计学意义（F=146．31，P〈0．05）。实验组G．期细胞的比例为52．2％±4．4％，明显低于空载体组的62．0％±5．O％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．O％和对照组的21．8％±1．3％，3组细胞G．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MDRl蛋白的表达量为1．854-0．18，高于空载体组的1．124-0．08和对照组的1．024-0．09，3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。
Objective To investigate the effects of over-expression of caudal-related homeodomain tran- scription 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-reeombi- nated 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 adriamyein, 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. Results The transfection efficiency was above 90% after transfectingthe SGC7901/DDP cells by lentiviral vector for 72 hours. The relative expression of Cdx2 protein in the experi- mental 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 ICs0 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 ＋0.05）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） rag/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 adriamyein in the experimental group was 18.20% _＋ 0. 12% , which was significantly higher than 7.22%＋0.09% of the empty vector group and 8.79% -＋0.11% of the control group （F = 146.31, P 〈0.05）. The proportion of ceils in the G1 phase in the experimental group was 52.2% ＋-4.4% , which was significantly lower than 62.0% ＋ 5.0% of the empty vector group and 67.8% -＋ 3.3% of the control group. The proportion of cells in the S phase in the experimental group was 40.4% ＋ 1.5% , which was significantly higher than 25.6% ＋ 2.0% of the empty vector group and 21.8% ＋- 1.3% of the control group. There were significant differences in the proportion of cells in the G1 phase and S phase between the 3 groups （ F = 17.08,236. 83, P 〈 0.05 ）. The cell apoptotic rate of the experimental group was 7.6%-＋ 1.3% , which was significantly lower than 11.1% ＋- 1.1% of the empty vector group and 10.8% ＋ 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 zastric cell line SGC7901/DDP.
出处
《中华消化外科杂志》
CAS
CSCD
北大核心
2014年第2期145-148,共4页
Chinese Journal of Digestive Surgery
基金
国家自然科学基金（81060201）
广西自然科学基金（2011GXNSFA018273）
重大疾病防治新技术研究开发项目（桂科攻1298003-2-6）
关键词
胃肿瘤
尾型同源盒基因2
多药耐药性
Gastric neoplasms
Caudal-related homeodomain transcription 2
Multi-drug resistance
