Objective: In order to evaluate potential application for diagnosis and prognosis of non-small cell-lung cancer (NSCLC), as well as to determine its role in the pathogenesis of the disease, we prepared anti-human h...Objective: In order to evaluate potential application for diagnosis and prognosis of non-small cell-lung cancer (NSCLC), as well as to determine its role in the pathogenesis of the disease, we prepared anti-human hnRNPA2/B1 potyclonal antibody. Methods: Prokaryotic expression vector of pET28a (+)-hnRNP A2/B1 was constructed and bansformed into E.coli BL21. The recombinant protein induced by IPTG was purified and injected to rabbits for antibody preparation. Expression of hnRN P A2/B1 was examined in 45 tissues of NSCLC and 16 inflammatory pseudotumor tissues of lung by immunohistochemistry with the antibody. The commercial hnRNP A2/B1 monoclonal antibody was used as a controI.Results: (1) Polyclonal an-tibody against hnRNP A2/B1 with high title was obtained. (2) The positive staining in NSCLC tissues was 62.22%, which was substantially higher than that in normal tissues (40%, P = 0.035) or inflammatory pseudotumor tissues (31.25%, P=0.033). (3) Expression of hnRNP A2/B1 positively correlated with age and the history of smoking, whereas it negatively correlated with differentiation staging of tumors. (4) Follow-up study showed that the survival time of patients with positive staining was significantly shorter than that of patients without hnRNP A2/B1 expression (P=0.048). Conclusion: It is successful to make the recombinant protein and prepare the polyclonal antibody agonist human hnRNP A2/B1. It may be a valuable marker for the diagnosis and prognosis of NSCLC. Our results provide a basis for further study in clinical application.展开更多
基金a grant from the Provincial Natural Sciences Foundation of Anhui(No.050430703)Major Scientific Research Programs of the Department of Science and Technology of Anhui(No.05023086)
文摘Objective: In order to evaluate potential application for diagnosis and prognosis of non-small cell-lung cancer (NSCLC), as well as to determine its role in the pathogenesis of the disease, we prepared anti-human hnRNPA2/B1 potyclonal antibody. Methods: Prokaryotic expression vector of pET28a (+)-hnRNP A2/B1 was constructed and bansformed into E.coli BL21. The recombinant protein induced by IPTG was purified and injected to rabbits for antibody preparation. Expression of hnRN P A2/B1 was examined in 45 tissues of NSCLC and 16 inflammatory pseudotumor tissues of lung by immunohistochemistry with the antibody. The commercial hnRNP A2/B1 monoclonal antibody was used as a controI.Results: (1) Polyclonal an-tibody against hnRNP A2/B1 with high title was obtained. (2) The positive staining in NSCLC tissues was 62.22%, which was substantially higher than that in normal tissues (40%, P = 0.035) or inflammatory pseudotumor tissues (31.25%, P=0.033). (3) Expression of hnRNP A2/B1 positively correlated with age and the history of smoking, whereas it negatively correlated with differentiation staging of tumors. (4) Follow-up study showed that the survival time of patients with positive staining was significantly shorter than that of patients without hnRNP A2/B1 expression (P=0.048). Conclusion: It is successful to make the recombinant protein and prepare the polyclonal antibody agonist human hnRNP A2/B1. It may be a valuable marker for the diagnosis and prognosis of NSCLC. Our results provide a basis for further study in clinical application.