The Expression Of CXCR7 In Esophageal Carcinoma And Its Effect On Cell Proliferation

BackgroundThe occurrence and development of malignant tumors is a complex process with multiple genes regulation in different stages,which involves the activation of oncogenes or the inactivation of anti-oncogenes.Various reasons lead to oncogenes expression increased or anti oncogene expression decreased will cause other molecules level changes in corresponding signal pathway,then further influence the biological behavior of cells.Studies have shown that chemokines and their receptors are closely related to tumors.CXCR7,as a chemokine receptor,can participate in the regulation of various malignancies after binding to ligand.However,the expression of CXCR7 and its role in esophageal cancer are poorly understood.ObjectiveTo determine the expression level of CXCR7 in esophageal cancer and therelationship between CXCR7 expression and esophageal cancer cell proliferation.Explore CXCR7 role in cancer development in order to find new molecular targets for esophageal cancer and provide a theoretical basis for clinical diagnosis and treatment.Methods1.Total RNA was extracted from 14 cases of esophageal cancer tissues and corresponding adjacent normal tissues,and then reverse transcription was used togenerate c DNA.The quantitative RT-PCR technique was used to detect the expression levels of CXCR7 mRNA in esophageal cancer tissues and their adjacent normal tissues.2.Wild type and mutation type of CXCR7 specificity primers were designed to amplify human wild type and mutant CXCR7 c DNAs by PCR technique,and then were subcloned into p EGFP-N1 to construct p EGFP-wt CXCR7 and p EGFP-CXCR7?C through double enzyme digestion and directional cloning,respectively.A small hairpin RNA targeting human CXCR7 gene was designed and synthesized as a complementary oligonucleotide chain,and was then ligated into the p Silencer4.1-CMV neo to construct the CXCR7 sh RNA expression plasmid-p Silencer4.1-sh CXCR7.3.The above prebuilt recombinant plasmids were transiently transfected into esophageal cancer cell line KYSE-510,respectively.Their effects on esophageal cancer cell biology were observed and photographed after48 hours.Western blot experiment was used to detect the expression of recombinant plasmid in esophageal squamous cell carcinoma.4.The above prebuilt recombinant plasmids were transiently transfected into esophageal cancer cell line KYSE-510,respectively.MTT assay was used to detect the optical density value of the experimental group cells and the control groupcells at 0h,24 h,48h and 72 h.Cell proliferation was analyzed.Results1.CXCR7 mRNA relative expression in esophageal cancer tissues(3.024�2.910)higher than that in adjacent normal tissues(1.446�1.118),the difference was statistically significant(P<0.05).2.The expression plasmids(pEGFP-wtCXCR7,p EGFP-CXCR7?C and p Silencer4.1-sh CXCR7)had been constructed successfully.The recombinant plasmids were able to successfully express in esophageal cancer cells after the transfection operation.3.MTT assay was used to detect the optical density value of each group cells at 24 h,48h and 72 h after transfection.Wild type group(0.663�0.032?1.120�0.032?1.343�0.037)and mutant type group(0.573�0.031?0.948�0.050?1.189�0.024)were higher than their control group(0.469�0.027?0.768�0.031?0.991�0.041),and wild type group is higher than mutant type group,the difference was statistically significant(P<0.01);Interference group(0.561�0.025 ? 1.012�0.026 ? 1.303�0.028)was higher than that of the control group(0.449�0.031 ? 0.748�0.033 ? 1.039�0.037)and the difference is statistically significant(P<0.01).Conclusion1.CXCR7 is highly expressed in esophageal cancer,suggesting that it is closely related to the occurrence of esophageal cancer.2.The up-regulation and down-regulation of CXCR7 expression were able to promote the proliferation of esophageal cancer cell,suggesting that it may play a regulatory role on tumor cells through a direct or indirect pathway.3.Wild-type CXCR7 plasmid showed stronger activity of promoting cancer cells proliferation,suggesting the c-terminal of CXCR7 plays an indispensable role in its gene function.