摘要
目的探讨趋化因子受体CXCR7在食管癌中的表达及对其细胞增殖的影响。方法应用实时定量聚合酶链反应(qRT-PCR)技术检测CXCR7在食管癌和癌旁组织中的表达;利用聚合酶链反应(PCR)技术扩增人CXCR7野生型和突变型cDNA全长,构建pEGFP-wtCXCR7和pEGFP-CXCR7ΔC质粒;设计合成靶向CXCR7基因的短发夹RNA(shRNA),构建CXCR7shRNA表达质粒pSilencer4.1-shCXCR7。将构建好的质粒分别转染人食管癌细胞KYSE-510,检测其对细胞增殖的影响。结果与癌旁组织比较,CXCR7在食管癌中高表达;成功构建了表达质粒pEGFPwtCXCR7、pEGFP-CXCR7ΔC和pSilencer4.1-shCXCR7;CXCR7表达水平上调和下调均可促进食管癌细胞KYSE-510的增殖。结论CXCR7可能直接或通过代偿途径促进食管癌细胞增殖,在食管癌发生、发展过程中可能发挥重要作用。
Objective To investigate the expression level of chemokine receptor CXCR7 in esophageal cancer and its effect on esophageal cancer cell proliferation. Methods The quantitative real time polymerase chain reaction (qRT-PCR) technique was used to detect the expression level of CXCR7 in esophageal cancer tissues and their adjacent normal tissues. Human wild type and mutant CXCR7 cDNAs were amplified by PCR technique, and then pEGFP-wtCXCR7 and pEGFP-CXCR7AC were contructed through double enzyme digestion and directional cloning, respectively. A small hairpin RNA(shRNA) targeting human CXCR7 gene was designed and synthesized as a complementary oligonucleotide chain, and the CXCR7 shRNA expression plasmid-pSilencer4.1-shCXCR7 was constructed. All the plasmids were transiently transfected into esophageal cancer cell line KYSE-510 cells. And its effect of the cell proliferation was analyzed. Results CXCR7 was overexpressed in esophageal cancer tissues compared to the adjacent normal tissues. The expression plasmids (pEGFP-wtCXCR7 ,pEGFP-CXCR7AC and pSilencer4.1-shCXCR7) had been constructed successfully. Both the increase and the decrease of CXCR7 were able to induce cell proliferation in KYSE-510 cells. Conclusion CXCR7 may promote esophageal cancer cell proliferation through a direct or compensatory pathway, thus, CXCR7 may play an important role in the development of esophageal cancer.
作者
王佳琪
陈志国
韩荣飞
鲍永华
张会娟
王倩
李凯
杨万才
WANG Jia-qi CHEN Zhi-guo HAN Rong-fei BAO Yong-hua ZHANG Hui-juan WANG Qian LI Kai YANG Wan-cai(Department of Pathology, Basic Medical College,Xinxiang Medical University, Xinxiang 453003, Henan Province, Chin)
出处
《新乡医学院学报》
CAS
2016年第1期16-19,共4页
Journal of Xinxiang Medical University
基金
国家自然科学基金项目资助(编号:81272251)
美中抗癌协会项目资助(编号:USCACA-TIGM-001)
关键词
CXCR7
食管癌
真核表达质粒
细胞增殖
CXCR7
esophageal cancer
eukaryotic expression plasmid
cell proliferation