| Background and objectiveFerroportin(FPN)as a key protein in iron metabolism pathway,has attracted much attention recently.Studies have shown that iron metabolism is associated with the occurrence and progression of non-small cell lung cancer(NSCLC).To investigate the role of FPN in NSCLC cell proliferation and migration,we established different FPN expression systems to compare the effects of FPN on the proliferation and migration ability of NSCLC cells.Methods From February 2018 to October 2018,80 patients underwent thoracic surgery in the first affiliated hospital of University of Science and Technology of China were collected,and the basic information and clinical materials of the patients were recorded,including gender,age,pathological type of lung cancer and TNM stage of the patients.After the operation,the samples were subjected to immunohistochemical treatment,and the final mean staining positive score(MPSS)was calculated based on the addition of the positive staining cell area percentage score and staining intensity score.The relationship between parameters and clinical histological type,pathological type andstage was analyzed by one-way anova and two independent samples t-test.Human normal bronchial epithelial cells HBE and NSCLC cells A549 were cultured.Cells were collected to make protein samples when 90% of the cells were grown in a petri dish,and then quantified using BCA.The same amount of protein samples were taken for western blot,and the electrophoretogram was analyzed by gray scale and t test.FPN overexpression system(p CDH-fpn-f)was constructed on empty PCDH vector,and the sh RNA library was queried to construct knock down system(sh FPN8,9).Lentivirus packaging was performed to infect NSCLC cell lines A549 and H1299,respectively.FPN expression in NSCLC cell lines was detected by western blot(WB)and real-time fluorescence quantitative PCR(qrt-pcr).The gray value of electrophoresis and PCR results were used for t test statistical analysis.NSCLC cell lines A549 and H1299 were cultured and infected with lentiviruses containing overexpressed and knock down systems,respectively.The effects of FPN at different expression levels on proliferation and migration of lung cancer cells were detected by scratch experiment and growth curve method after the cells were overgrown in petri dishes.Results IHC showed that MPSS of FPN in 20 normal lung tissues was 4.60±1.23,and in60 cancer tissues was 2.42±0.83,and the expression of FPN was associated with tumor(P=0.000).Among the 60 cancer tissue samples,MPSS of FPN in 19 SCC cases was2.58±0.16,in 29 ADC cases was 2.36±1.50,and the expression of FPN was not correlated with pathological type(P=0.354).In TNM stage,MPSS of FPN in 2 patients with stage I was 3.00±1.41,in 19 patients with stage II was 2.53±1.07,in 24 patients with stage III was 2.21±0.72,and in 15 patients with stage IV was 2.53±0.52,among which the expression of FPN was not significantly correlated with clinical stage(P=0.360).In cell experiments,compared with HBE,the expression level of FPN in A549 decreased significantly.Both WB and qrt-pcr showed that p CDH-fpn-f up-regulated the expression of FPN in lung cancer cells,while sh FPN down-regulated the expression of FPN.The high expression of FPN inhibited the proliferation and migration of lung cancer cells,while the low expression was the opposite.ConclusionThe expression of FPN in NSCLC was down-regulated by IHC detection of clinical tissue samples and WB detection of lung cancer cell lines.Further regulation of the expression level of FPN can lead to changes in the biological behavior of lung cancer cells.Specifically,the down-regulation of FPN expression level can promote the proliferation and migration ability of cancer cells,while the up-regulation of FPN expression level can inhibit the proliferation and migration ability of cancer cells.Detecting the expression level of FPN can predict or reflect lung cancer from a new perspective,which has certain value for the evaluation of prevention or prognosis in patients. |
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