The Effect Of CXCL12-CXCR4 Biological Axis On The Ability Of Proliferation, Migration And Invasion Of Hela Cells

Objective: Human Cervical cancer(HCC)is the most common malignant tumors in women.Recently;age of onset has been in advance.The molecular mechanism of occurrence and development of HCC is still not exactly clear.Tumor cell proliferation,invasion and metastasis are the main biological action,and it is also the leading cause of death in patients with malignant tumor.HCC is complex,continuous and multistage process,demonstrating highly tissue-specific.The most common route of metastasis is to the place promenade and adjacent tissues and lymph node,while hematogenous metastasis is less.The invasion of HCC is different at different stages.Tumor cells metastasis to organ-special produce a variety of mechanisms to promote the invasion of tissue,stimulating vascular or lymphatic vessel formation,producing a variety of cytokines,creating conditions for tumor metastasis.In recent research of the “signal netwoks”study,the CXCL12-CXCR4 biological axis consist of chemokine CXCL12 and its specific receptor CXCR4 plays an important role in the development of all kinds of tumors,and we also found the high express of CXCR4 in HCC tissue.In this research,we cultured HCC cell strains Hela cells in vitro,and observed its growth features and HE dyeing to study cell strcture,and investigated the expression of CXCR4 protein in HeLa cell by immunocytochemistry.Then,MTT was used to analyze the Hela cells proliferation ability after treatment with different concentrations of CXCLl2.Finally,on serum-free culture conditions,transwell method was used to detect the changes of Hela cells migration and invasion ability after treatment with different concentrations of CXCLl2 and to determine inhibition of CXCR4 antagonist AMD3100.This experiment to investigate the biologic axis CXCL12-CXCR4 in HCC provides theoretical basis,and also demonstrate the specific blocking of CXCR4 antagonist AMD3100,indicating that interaction of CXCR4 and CXCL12 might provide new treatment of HCC patients.CXCL12-CXCR4 might become new targets for anticancer treatment.Methods:1 Hela was cultured in vitro.When cell density reached 80%,cytoarchitecture was observed by optical microscopy and HE staining.2 To investigate the expression of CXCR4 protein in Hela cell by immunocytochemistry.3 MTT was used to analyze the Hela cells proliferation ability after treatment with different concentrations of CXCLl2.4 On serum-free culture conditions,transwell method was used to detect the changes of Hela cells migration and invasion ability after treatment with different concentrations of CXCLl2 and to determine inhibition of CXCR4 antagonist AMD3100.Results:1 Hela cells were polygonal,similar to epithelial cells,when cultured in vitro;Hela cells nuclei were large,dark by HE stain,some cells contained many nuclei,and cytoplasm were rich.2 CXCR4 strong staining is showed in Hela cells cytomembrance and cytoplasm by immunocytochemistry.It was the evidence that CXCR4 was expressed both in cytomembrance and in cytoplasm.3 MTT disclosed that Hela cells showed stronger proliferation ability with concentrations of 10 mg/mlCXCL12 、 100mg/ml CXCL12 vs.blank group((0.52±0.06)vs.(0.41±0.02),(0.66±0.07)vs.(0.41±0.02)),(P<0.05).Beside s,CXCR4 antagonist AMD3100 could inhibit Hela cells proliferation((0.66±0.07)vs.(0.43±0.04)),(P<0.05).4(1)Under serum-free suboptimal condition,10mg/ml CXCL12 can promote cell migration((166.80±4.84)vs.(136.87±5.28)),100mg/ml CXCL12 can significantly promote cell migration(223.20±8.51,P<0.05).This enhancing effect of CXCL12 on cell migration is increased with increasing concentration of CXCL12,and is strongly inhibited by treatment with 10μg/ml CXCR4 inhibitor AMD3100(82.87±4.36,P<0.05).(2)Matrigel is reconstituted artificial basal membrane,which have the shape and function of internal basal membrane.Under serum-free suboptimal condition,Hela displayed minimal invasiveness through Matrigel when CXCL12 was no present in the lower chamber of transwell(32.93± 1.94).10mg/ml CXCL12 promoted cell invasion across Matrigel(37.00±2.14,P<0.05).The number of invading cells through Matrigel in the presence of 100mg/ml CXCL12 was significantly higher than that of 10mg/ml CXCL12(57.40±1.64,P<0.05).The enhancing effect of CXCL12 on cell invasion was increased with increasing concentration of CXCL12,and was strongly inhibited by treatment with 10μg/ml CXCR4 inhibitor(24.67±2.19,P<0.05),which proved AMD3100 could inhibit CXCL12-CXCR4 biological axis function.Conclusions:1 When cultured in vitro,Hela cells were in good shape of polygon or fusiformis.2 CXCR4 was strongly expressed both in cytomembrance and in cytoplasm by immunocytochemistry.3 CXCL12-CXCR4 biological axis could enhance the ability of proliferation,migration and invasion of Hela cells,and AMD3100 could inhibit the CXCR4 positive tumor cells proliferation,migration and invasion by inhibiting CXCL12-CXCR4 biological axis function.4 Inhibition of CXCR4 function could prevent tumor cells proliferation,migration and invasion so that combined with CXCR4 antagonist was new therapy target for cervical cancer.It was of potential clinical significance.