AMD3100对胆管癌CXCR4基因、蛋白表达及QBC939细胞增殖能力的影响-东南大学学报医学版

AMD3100对胆管癌CXCR4基因、蛋白表达及QBC939细胞增殖能力的影响

单位：1. 长沙医学院附属株洲市人民医院普通外科, 湖南株洲 412000;
2. 南华大学附属南华医院普通外科, 湖南衡阳 421002

分类号：R735.8

出版年·卷·期（页码）：2019·38·第一期（98-102）

摘要：

目的：探讨CXCR4基因在胆管癌组织标本及人胆管癌细胞株QBC939中的表达情况，分析AMD3100应用对CXCR4基因表达及QBC939细胞的生物学行为变化。方法：收集人胆管癌组织标本，培养人胆管癌细胞株QBC939，加入不同浓度的AMD3100抑制剂。采用RT-PCR和Western blot方法，分别检测人胆管癌标本及转染AMD3100抑制剂前后QBC939胆管癌细胞株中CXCR4基因和蛋白的表达。应用CCK-8试剂盒对转染后细胞增殖活性进行检测。通过克隆形成实验观察AMD3100对细胞克隆形成的抑制。结果：CXCR4基因及其蛋白在胆管癌组织标本及人胆管癌细胞株QBC939中均高度表达。不同浓度AMD3100抑制剂对QBC939胆管癌细胞株的增殖抑制活性取决于AMD3100抑制剂的孵育浓度、孵育时间。其中，转染不同浓度抑制剂24 h，并未对细胞增殖造成影响；转染48 h后，0.1 μg·ml^-1AMD3100无明显影响，1 μg·ml^-1、5 μg·ml^-1的AMD3100抑制剂明显抑制了细胞的增殖；转染72 h后，各浓度均对细胞增殖有明显抑制作用。克隆形成实验分析显示，抑制CXCR4基因表达明显延长了克隆体形成时间，使克隆体体积和数目减少。结论：AMD3100抑制剂影响人胆管癌细胞株QBC939的增殖能力，抑制CXCR4基因和蛋白的表达。

Objective:To investigate the expression of CXCR4 gene in cholangiocarcinoma tissue and human cholangiocarcinoma cell line QBC939. The effects of AMD3100 on CXCR4 gene expression and biological behavior of QBC939 cells were analyzed. Methods:Human cholangiocarcinoma tissue specimens were collected, and human cholangiocarcinoma cell line QBC939 was cultured and different concentrations of AMD3100 inhibitors were added. RT-PCR and Western blot methods were used to detect the expression of CXCR4 gene and protein in human cholangiocarcinoma specimens and QBC939 cholangiocarcinoma cell lines before and after transfection with AMD3100 inhibitor. The CCK-8 kit was used to detect the cell proliferation activity after transfection. The inhibition of cell colony formation with AMD3100 was observed by clone formation experiments. The data were analyzed using SPSS 19.0 software. Results:The CXCR4 gene and its protein were highly expressed in cholangiocarcinoma tissue samples and human cholangiocarcinoma cell line QBC939. Different concentrations of AMD3100 inhibitors had different effects on the proliferation of QBC939 cholangiocarcinoma cell lines depending on the incubation concentration and incubation time of AMD3100 inhibitors. however, transfection with different concentrations of inhibitors for 24 h did not affect cell proliferation. After 48 hours of transfection, 0.1 μg·ml^-1 AMD3100 had no significant effect, and 1 μg·ml^-1 and 5 μg·ml^-1 of AMD3100 inhibitor significantly inhibited cell proliferation. After 72 hours of transfection, each concentration had a significant inhibitory effect on cell proliferation. Analysis of clone formation experiments showed that inhibition of CXCR4 gene expression significantly prolonged the formation of clones and reduced the size and number of clones. Conclusion:AMD3100 inhibitors affect the proliferation of human cholangiocarcinoma cell line QBC939 and inhibit the expression of CXCR4 gene and protein.

参考文献：

[1] 李昌,杨岩,蓝诺,等.肝外胆管癌组织MMP-2表达及其临床意义[J].中华肿瘤防治杂志,2017,24(10):689-691.
[2] 魏金鑫,刘胜利.肝门部胆管癌术前肝功能维护的几个热点问题[J].东南大学学报:医学版,2016,35(5):778-780.
[3] 胡剑飞,谢建平.不同手术方法治疗肝内胆管癌的临床效果评价及预后危险因素分析[J].实用癌症杂志,,2018,33(3):408-411.
[4] 马达,丁佑铭.肝细胞癌合并胆管癌的临床分型及手术方式对其远期疗效的影响[J].实用癌症杂志,2017,32(1):121-123.
[5] SILSIRIVANIT A,SAWANYAWISUTH K,RIGGINS G J,et al.Cancer biomarker discovery for cholangiocarcinoma:the high-throughput approaches[J].J Hepatobiliary Pancreat Sci,2014,21(6):388-396.
[6] 苏德望,付先东,于佳楠,等.血清糖类抗原19-9、癌胚抗原在胆管癌诊断中的应用价值[J].中国实验诊断学,2017,21(10):1693-1695.
[7] 唐映,陈浩军,林树洪.ERCP胆管刷检联合血清、胆汁中CA19-9、CEA对胆管良、恶性狭窄鉴别诊断的价值[J].现代消化及介入诊疗,2017,22(1):48-51.
[8] 吴唯,钱立元,戴荆,等.不同分子分型乳腺癌组织中趋化因子CXCL12及其受体CXCR4和CXCR7的表达及临床意义[J].中南大学学报:医学版,2017,42(2):147-153.
[9] 王振,辛明明,杨德林.CXCL12/CXCR4生物学与膀胱肿瘤关系[J].医学综述,2012,18(6):843-846.
[10] 刘静,郭华雄.趋化因子CXCL12/CXCR4生物轴在肿瘤中的研究进展[J].临床与实验病理学杂志,2010,26(4):477-480.
[11] 杨春康,简陈兴,应敏刚.CXCR4抑制剂AMD3100对结肠癌肝转移的实验研究[J].中华肿瘤防治杂志,2010,17(18):1422-1424.
[12] 霍胜军,汤恢焕,曾祥福.CXCR4抑制剂LPS对胆管癌细胞株QBC939生物学行为的影响[J].中国肿瘤临床,2010,37(12):669-672.
[13] 张引,余力信,张发鹏,等.趋化因子受体CXCR4及其配体CXCL12在肝癌侵袭转移中的作用及机制[J].天津医药,2018,46(1):20-26.
[14] 张卉,张树才.非小细胞肺癌EGFR基因靶向治疗研究进展[J].中国肺癌杂志,2017,20(1):61-65.
[15] 孔凡妮,查剑英,杨圆圆,等.以脐带间充质干细胞为载体的靶向基因治疗系统联合5-FU对裸鼠HepG2移植瘤的作用[J].山东医药,2017,57(26):9-12.
[16] 尹东,张娜,张佐,等.CXCR4拮抗剂AMD3100对人口腔鳞状细胞癌细胞增殖和凋亡的影响[J].宁夏医学杂志,2016,38(10):879-881.
[17] 王士猛.下调TCF3抑制非小细胞肺癌细胞的增殖和迁移能力[J].中国病理生理杂志,2017,33(2):289-296.

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